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Sample records for bombyx mori identified

  1. Autophagy studies in Bombyx mori

    Directory of Open Access Journals (Sweden)

    L Tian

    2015-03-01

    Full Text Available Autophagy, which is well conserved from yeast to mammals, plays essential roles in development and diseases. Using the domesticated silkworm, Bombyx mori, as a model insect, several reports on autophagy have been made recently. Autophagic features are observed in the midgut and fat body during the larval-pupal transition as well as the silk gland and ovarian nurse cells during the pupal stage. There are 14 autophagy related (Atg genes, including at least two transcript variants of Atg1, predicated in Bombyx. Expression of most Atg genes is consistent with the autophagy process in the fat body during the larval-pupal transition, and reduction of Atg1 expression by RNAi blocks this process. The molting hormone, 20-hydroxyecdysone (20E, and starvation induce autophagy in the fat body by upregulating Atg gene expression and blocking the PI3K-TORC1 pathway. Meanwhile, autophagy precedes apoptosis in the midgut and other larval tissues during the larval-pupal transition, while the detailed mechanism is not illustrated yet. We assume that there are at least four future directions about autophagy studies in Bombyx during the next years: (1 physiological functions of autophagy; (2 identification of new components involved in the autophagy process; (3 detailed molecular mechanism of autophagosome formation; (4 functional relationship between autophagy and apoptosis.

  2. PHYTOCHEMICAL INVESTIGATION OF THE SILK COCOONS OF BOMBYX MORI L.

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    Kaskoos Raad A.

    2012-05-01

    Full Text Available Silk cocoons, produced by Bombyx mori L. (Bombicidae are useful as hypotensive, expectorant, bronchodilator and attenuant drug in traditional medicine. Phytochemical investigation of the ethanolic extract of the cocoons led to the isolation of new phenolic constituents identified as n-butyl-3,4-dihydroxybenzoate (1, 3′,8′,9′-trigeranilanyl-3,4-dihydroxybenzoate (2, 3′,7′-dimethyl-3′-hydroxy-octanyl gallate (3, 3,4-dihydroxyphenyl-n-pentanyl ether (4 and 2,3,4-trihydoxypenyl-n-pentanyl ether (5 on the basis of spectral data analysis.

  3. Molecular Detection of Healthiness of Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    2012-01-01

    [Objective] The paper was to explore the regularity between heat shock protein expression and the healthiness changes of Bombyx moil materials. [Method] The representative heat shock protein gene Bmhsp24.3 was screened by bioinfor- matic analysis method, and carried out real-time PCR expression analysis. [Result] The target gene Bmhsp24.3 expressed in different B. mori materials, but the expres- sion level in different materials significantly varied. The relative expression level of the gene had different degrees of changes under different rearing conditions. With the increase of rearing temperature, the gene expression was upregulated. The ma- terials with better healthiness had remarkable increase in expression of target gene, while the materials with poorer healthiness had less increase in expression of target gene. The expression difference of target gene Bmhsp24.3 was exactly consistent with the healthiness of breeds. [Conclusion] The healthiness of materials had rela- tionship with expression of target gene Bmhsp24.3. the higher the expression of tar- get gene Bmhsp24.3 was, the better the healthiness of materials was; conversely, the lower the expression of target gene Bmhsp24.3 was, the poorer the healthiness of materials was.

  4. Transgenic breeding of anti-Bombyx mori L. nuclear polyhedrosis virus silkworm Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    Huijuan Yang; Wei Fan; Hao Wei; Jinwei Zhang; Zhonghua Zhou; Jianying Li; Jianrong Lin; Nong Ding; Boxiong Zhong

    2008-01-01

    Silkworm strains resistant to Bombyx mori L. nuclear polyhedrosis virus were obtained through transgenic experiments.piggyBac transposon with an A3 promoter were randomly inserted into the silkworm, driving the enhanced green fluorescent protein (EGFP) reporter gene into the silkworm genome. Polymerase chain reaction results verified the insertion of the extraneous EGFP gene, and fluorescence microscopy showed that the EGFP was expressed in the midgut tissue. The morbidity ratio of the nuclear polyhedrosis decreased from 90% in the original silkworm strain to 66.7% in the transgenic silkworm strain. Compared with the resistance to the Bombyx mori L. nuclear polyhedrosis virus in the Qiufeng strain, which is commonly used in the production, there was an increase of 33 centesimal points in the transgenic silkworms. The antivirotic character in the ChunhuaxQiuyue strain, which was bred from a different transgenic family, was about 10 centesimal points higher than that in the Qiufeng×Baiyu, another crossbreed used in production. Our results indicated a good application value of the transposon-inserted mutation in the breeding of anti-BmNPV silkworm strain.

  5. Comparative Proteome Analysis of Multi-Layer Cocoon of the Silkworm, Bombyx mori

    OpenAIRE

    Zhang, Yan; ZHAO, PING; Dong, Zhaoming; Wang, Dandan; Guo, Pengchao; Guo, Xiaomeng; Song, Qianru; Zhang, Weiwei; Xia, Qingyou

    2015-01-01

    Bombyx mori cocoon has a multi-layer structure that provides optimal protection for silkworm pupa. Research on the mechanical properties of the multi-layer structure revealed structure-property relationships of the cocoon. Here, we investigated the protein components of the B. mori cocoon in terms of its multi-layer structure. Liquid chromatography-tandem mass spectrometry identified 286 proteins from the multiple cocoon layers. In addition to fibroins and sericins, we identified abundant pro...

  6. Diapause Prevention Effect of Bombyx mori by Dimethyl Sulfoxide

    OpenAIRE

    Takayuki Yamamoto; Keisuke Mase; Hiroshi Sawada

    2013-01-01

    HCl treatment has been, for about 80 years, the primary method for the prevention of entry into embryonic diapauses of Bombyx mori. This is because no method is as effective as the HCl treatment. In this study, we discovered that dimethyl sulfoxide (DMSO) prevented entry into the diapause of the silkworm, Bombyx mori. The effect of diapause prevention was 78% as a result of treatment with 100% DMSO concentration, and the effect was comparable to that of the HCl treatment. In contrast, in the ...

  7. The polymorphism of sericin 2 gene in silkworm (Bombyx mori)

    OpenAIRE

    TYLLEROVÁ, Helena

    2010-01-01

    In our study, we examined the polymorphism of gene Ser2 from domesticated silkmoth Bombyx mori and its closest wild relative B. mandarina. As a starting material for our work, we used the restriction map of allele C isolated from hybrid lineages 200 and 300 of European silkmoth B. mori (Michaille et al. 1990a). We also used the published sequence of allele D which was isolated from {\\clq}qDaizo`` p50 strain of B. mori (Kludkiewicz et al 2009). Based on the published sequence, we designed PCR ...

  8. Retrotransposon "Qian" mediated segmental duplication in silkworm, Bombyx mori.

    Science.gov (United States)

    Xu, Yunmin; Jiang, Ning; Zou, Ziliang; Tu, Zhijian; Chen, Anli; Zhao, Qiaoling; Xiang, Zhonghuai; He, Ningjia

    2014-03-01

    Transposable elements constitute a large fraction of the eukaryotic genomes. They have the potential to alter genome structure and play a major role in genome evolution. Here, we report a segmental duplication mediated by a novel long terminal repeat (LTR) retrotransposon as the cause of an egg-shell recessive lethal mutant (l-em mutant) in silkworm (Bombyx mori). The segmental duplication resulted in the duplication of six genes and the disruption of two genes. Disruption of BmEP80 (B. mori egg protein 80), a gene encoding a major egg-shell structure protein, is likely responsible for the lethal water-loss phenotype in the l-em/l-em mutant. Our data revealed that BmEP80 is present in the inner egg-shell layer and plays important roles in resistance to water efflux form eggs. A novel LTR retrotransposon (named as "Qian") was identified and the model for the Qian-mediated chromosomal segmental duplication was proposed. Detail biochemical and genomic analyses on the l-em mutant offer an opportunity to demonstrate that an LTR retrotransposon could trigger duplication of a chromosomal segment (∼96.3 kb) and confer novel phenotype. PMID:24462715

  9. Efficient TALEN construction for Bombyx mori gene targeting

    Czech Academy of Sciences Publication Activity Database

    Takasu, Y.; Sajwan, Suresh; Daimon, T.; Osanai-Futahashi, M.; Uchino, K.; Sezutsu, H.; Tamura, T.; Žurovec, Michal

    2013-01-01

    Roč. 8, č. 9 (2013), e73458. E-ISSN 1932-6203 R&D Projects: GA ČR GAP305/10/2406 Grant ostatní: Japan Society for the Promotion of Science(JP) 23580083 Institutional support: RVO:60077344 Keywords : Bombyx mori Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.534, year: 2013 http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0073458

  10. Isolation and identification of a pathogen of silkworm Bombyx mori.

    Science.gov (United States)

    Tao, Heng-Ping; Shen, Zhong-Yuan; Zhu, Feng; Xu, Xiao-Fang; Tang, Xu-Dong; Xu, Li

    2011-03-01

    A pathogenic bacterial strain, ST-1, was isolated from a naturally infected silkworm. The strain was identified on the basis of its physiological and biochemical properties and the results of sequence analysis of its 16S rRNA gene. The results of the 16S rRNA gene sequence analysis revealed that ST-1 shared the highest sequence identity (more than 99%) with Pseudomonas chlororaphis subsp. aurantiaca. ST-1 bacteria were gram-negative and 0.7-0.9 × 1.3-1.5 μm long, short rods with rounded ends. The strain could utilize sodium citrate, malonate, D-glucose, sucrose, D-fructose, D-mannose, and L-arabinose. Pathogenicity of ST-1 for silkworm could be depicted as a linear regression of the logarithm (y) of ST-1 concentration against probability (x) (y = 0.4040 + 0.0600x). The median lethal concentration (LC(50)) was 2.12 × 10(4) cfu/ml. In conclusion, ST-1 was identified as Ps. chlororaphis subsp. aurantiaca. This is the first report that Ps. aurantiaca is a pathogen for silkworm Bombyx mori. PMID:21046395

  11. Surface display and bioactivity of Bombyx mori acetylcholinesterase on Pichia pastoris

    Science.gov (United States)

    To construct the Pichia pastoris (P. pastoris) cell surface display system of Bombyx mori acetylcholinesterase (BmAChE), the gene for the anchor protein (AGa1) was obtained from Saccharomyces cerevisiae and was fused with the modified Bombyx mori acetylcholinesterase gene (bmace) and transformed int...

  12. Precocious Metamorphosis in the Juvenile Hormone–Deficient Mutant of the Silkworm, Bombyx mori

    OpenAIRE

    Daimon, Takaaki; Kozaki, Toshinori; Niwa, Ryusuke; Kobayashi, Isao; Furuta, Kenjiro; Namiki, Toshiki; Uchino, Keiro; Banno, Yutaka; Katsuma, Susumu; Tamura, Toshiki; Mita, Kazuei; Sezutsu, Hideki; Nakayama, Masayoshi; Itoyama, Kyo; Shimada, Toru

    2012-01-01

    Insect molting and metamorphosis are intricately governed by two hormones, ecdysteroids and juvenile hormones (JHs). JHs prevent precocious metamorphosis and allow the larva to undergo multiple rounds of molting until it attains the proper size for metamorphosis. In the silkworm, Bombyx mori, several “moltinism” mutations have been identified that exhibit variations in the number of larval molts; however, none of them have been characterized molecularly. Here we report the identification and ...

  13. Alternative splicing and trans-splicing events revealed by analysis of the Bombyx mori transcriptome

    OpenAIRE

    Shao, Wei; Zhao, Qiong-Yi; Wang, Xiu-Ye; Xu, Xin-Yan; Tang, Qing; Li, Muwang; Li, Xuan; Xu, Yong-Zhen

    2012-01-01

    Alternative splicing and trans-splicing events have not been systematically studied in the silkworm Bombyx mori. Here, the silkworm transcriptome was analyzed by RNA-seq. The authors identified 320 novel genes, modified 1140 gene models, and found thousands of alternative splicing and 58 trans-splicing events. Studies of three SR proteins show that both their alternative splicing patterns and mRNA products are conserved from insect to human, and one isoform of Srsf6 with a retained intron is ...

  14. Identification of a putative RNAse III (dicer homolog gene in silkworm Bombyx mori

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    KM Ponnuvel

    2007-02-01

    Full Text Available Like other invertebrates, silkworms also encounter a problem from microbial infection including from RNA viruses. In insects, RNA interference acts as a natural anitiviral response to RNA virus infection. Especially in Drosophila, it is proved that dicer mediated RNA interference directs innate immunity against RNA viruses. This information prompted us to identify similar RNAse III (dicer gene in mulberry silkworm Bombyx mori. The Drosophila dicer gene was BLAST-searched with B. mori genome and single contig (GenBank accession n° AADK01001038 showed maximum homology with dicer gene, through which the RNAse III gene sequence was identified in the genome of silkworm B. mori. The RNAse III domain was present in the three regions with the length of 278 bp, 277 bp and 185 bp in the contig, possibly these three regions form exons. The primers were designed for three B. mori RNAse III regions and amplified through PCR. The region I was amplified only in pure Mysore silkworm strain whereas all three regions were amplified in Daizo strain. The PCR product sequences were translated and showed RNAse III domain with in the amplified product. The predicted B. mori RNAse III domain had phylogenetic relationship with other insect dicer genes. We presume that this RNAse III (dicer would protect B. mori larvae from invading RNA viruses, which exists in the other insects.

  15. Annotation and expression of carboxylesterases in the silkworm, Bombyx mori

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    Li Wen-Le

    2009-11-01

    Full Text Available Abstract Background Carboxylesterase is a multifunctional superfamily and ubiquitous in all living organisms, including animals, plants, insects, and microbes. It plays important roles in xenobiotic detoxification, and pheromone degradation, neurogenesis and regulating development. Previous studies mainly used Dipteran Drosophila and mosquitoes as model organisms to investigate the roles of the insect COEs in insecticide resistance. However, genome-wide characterization of COEs in phytophagous insects and comparative analysis remain to be performed. Results Based on the newly assembled genome sequence, 76 putative COEs were identified in Bombyx mori. Relative to other Dipteran and Hymenopteran insects, alpha-esterases were significantly expanded in the silkworm. Genomics analysis suggested that BmCOEs showed chromosome preferable distribution and 55% of which were tandem arranged. Sixty-one BmCOEs were transcribed based on cDNA/ESTs and microarray data. Generally, most of the COEs showed tissue specific expressions and expression level between male and female did not display obvious differences. Three main patterns could be classified, i.e. midgut-, head and integument-, and silk gland-specific expressions. Midgut is the first barrier of xenobiotics peroral toxicity, in which COEs may be involved in eliminating secondary metabolites of mulberry leaves and contaminants of insecticides in diet. For head and integument-class, most of the members were homologous to odorant-degrading enzyme (ODE and antennal esterase. RT-PCR verified that the ODE-like esterases were also highly expressed in larvae antenna and maxilla, and thus they may play important roles in degradation of plant volatiles or other xenobiotics. Conclusion B. mori has the largest number of insect COE genes characterized to date. Comparative genomic analysis suggested that the gene expansion mainly occurred in silkworm alpha-esterases. Expression evidence indicated that the expanded

  16. Tuning Molecular Weights of Bombyx mori (B. mori) Silk Sericin to Modify Its Assembly Structures and Materials Formation

    OpenAIRE

    Yang, Mingying; Shuai, Yajun; Zhou, Guanshan; Mandal, Namita; Zhu, Liangjun; Mao, Chuanbin

    2014-01-01

    Bombyx mori (B. mori) silk sericin is a protein with features desirable as a biomaterial, such as increased hydrophilicity and biodegradation, as well as resistance to oxidation, bacteria, and ultraviolet light. In contrast to other widely studied B. mori silk proteins such as fibroin, sericin is still unexplored as a building block for fabricating biomaterial, and thus a facile technique of processing it into a material is needed. Here, electrospinning technology was used to fabricate it int...

  17. Distinctive presence of peritracheal athrocytes in Bombyx mori L. and Bombyx mandarina M. as compared to their absence in several other lepidopteran species.

    Science.gov (United States)

    Owa, Chie; Aoki, Fugaku; Nagata, Masao

    2006-06-01

    Pericardial cells are present in a wide variety of insects and are thought to constitute the majority of 'athrocytes (nephrocytes)'. In contrast, peritracheal athrocytes have only been observed in Bombyx mori L. Although peritracheal athrocytes have a distinct morphology, it is unknown whether these cells are common to all lepidopterans. We anatomically compared eight lepidopteran species: Bombyx mori L. and Bombyx mandarina M. (Bombycidae); Samia cynthia ricini D. (Saturniidae); Agrius convolvuli L. (Sphingidae); Spodoptera litura F. and Mythimna separata W. (Noctuidae); Pieris rapae L. (Pieridae); and Glyphodes pyloalis W. (Crambidae). Of these species, only Bombyx mori L. and Bombyx mandarina M. possess peritracheal athrocytes. PMID:18089062

  18. Enhancer activity of Helitron in sericin-1 gene promoter from Bombyx mori.

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    Huang, Ke; Li, Chun-Feng; Wu, Jie; Wei, Jun-Hong; Zou, Yong; Han, Min-Jin; Zhou, Ze-Yang

    2016-06-01

    Sericin is a kind of water-soluble protein expressed specifically in the middle silk gland of Bombyx mori. When the sericin-1 gene promoter was cloned and a transgenic vector was constructed to express a foreign protein, a specific Helitron, Bmhel-8, was identified in the sericin-1 gene promoter sequence in some genotypes of Bombyx mori and Bombyx mandarina. Given that the Bmhel-8 Helitron transposon was present only in some genotypes, it could be the source of allelic variation in the sericin-1 promoter. The length of the sericin-1 promoter sequence is approximately 1063 or 643 bp. The larger size of the sequence or allele is ascribed to the presence of Bmhel-8. Silkworm genotypes can be homozygous for either the shorter or larger promoter sequence or heterozygous, containing both alleles. Bmhel-8 in the sericin-1 promoter exhibits enhancer activity, as demonstrated by a dual-luciferase reporter system in BmE cell lines. Furthermore, Bmhel-8 displays enhancer activity in a sericin-1 promoter-driven gene expression system but does not regulate the tissue-specific expression of sericin-1. PMID:27067405

  19. Cloning and Alternative Splicing Analysis of Bombyx mori Transformer-2 Gene using Silkworm EST Database

    Institute of Scientific and Technical Information of China (English)

    Bao-Long NIU; Zhi-Qi MENG; Yue-Zhi TAO; Shun-Lin LU; Hong-Biao WENG; Li-Hua HE; Wei-Feng SHEN

    2005-01-01

    We have identified Bombyx mori transformer-2 gene (Bmtra-2) cDNA by blasting the EST database of B. mori. It was expressed in the whole life of the male and female silkworm and was observed as a band of 1.3 kb by Northern blot analysis. By comparing corresponding ESTs to the Bmtra-2 DNA sequence,it was revealed that there were eight exons and seven introns, and all splice sites of exons/introns conformed to the GT/AG rule. Bmtra-2 pre-mRNA can produce multiple mRNAs encoding six distinct isoforms of BmTRA-2 protein using an alternative splicing pathway during processing. Six types of Bmtra-2 cDNA clones were identified by reverse transcription-polymerase chain reaction. All isoforms of BmTRA-2 protein contain two arginine/serine-rich domains and one RNA recognition motif, showing striking organizational similarity to Drosophila TRA-2 proteins.

  20. Biochemical characterization of rab proteins from Bombyx mori.

    Science.gov (United States)

    Uno, Tomohide; Moriwaki, Tsubasa; Nakamura, Masahiko; Matsubara, Mamoru; Yamagata, Hiroshi; Kanamaru, Kengo; Takagi, Michihiro

    2009-02-01

    The small GTPases known as Rab proteins are key regulators of membrane trafficking. We used RT-PCR to isolate cDNA clones of insect-specific Rab proteins (BRabN1 and BRabN2) showing low homology with known Rab proteins from other animals, from mRNA of Bombyx mori. These 2 Rabs were produced in Escherichia coli and purified. BRabN1 bound [(3)H]-GDP and [(35)S]-GTPgammaS with dissociation constants of 0.087 x 10(-6) M and 1.02 x 10(-6) M, respectively, whereas those of BRabN2 were 0.546 x 10(-6) M and 1.02 x 10(-6) M, respectively. Binding of [(35)S]-GTPgammaS to BRabN1 and N2 was inhibited by GDP and GTP. The GTP-hydrolysis activities of BRabN1 and N2 were 154 and 35.5 mmol/min/mole, respectively, and bound [(35)S]-GTPgammaS was exchanged efficiently with GTP. BRabN1 also showed ATPase activity and exchange of [(35)S]-GTPgammaS with ATP. Monoclonal antibodies against BRabN1 and N2 did not recognize any other Rab proteins, and Western blotting using the anti-BRabN1 antibody revealed a single band in the testis of B. mori. These results suggest that BRabN1 and N2 of B. mori bind GTP, convert from the GTP-bound state to the GDP-bound state by intrinsic GTP hydrolysis activity, and return to the GTP-bound state with the exchange, and that BRabN1 is specifically expressed in testis. Arch. Insect Biochem. Physiol. 2008. (c) 2008 Wiley-Liss, Inc. PMID:18949803

  1. Sialylation potentials of the silkworm, Bombyx mori; B. mori possesses an active α2,6-sialyltransferase.

    Science.gov (United States)

    Kajiura, Hiroyuki; Hamaguchi, Yuichi; Mizushima, Hiroki; Misaki, Ryo; Fujiyama, Kazuhito

    2015-12-01

    N-Glycosylation is an important post-translational modification in most secreted and membrane-bound proteins in eukaryotic cells. However, the insect N-glycosylation pathway and the potentials contributing to the N-glycan synthesis are still unclear because most of the studies on these subjects have focused on mammals and plants. Here, we identified Bombyx mori sialyltransferase (BmST), which is a Golgi-localized glycosyltransferase and which can modify N-glycans. BmST was ubiquitously expressed in different organs and in various stages of development and localized at the Golgi. Biochemical analysis using Sf9-expressed BmST revealed that BmST encoded α2,6-sialyltransferase and transferred N-acetylneuraminic acid (NeuAc) to the nonreducing terminus of Galβ1-R, but exhibited the highest activity toward GalNAcβ1,4-GlcNAc-R. Unlike human α2,6-sialyltransferase, BmST required the post-translational modification, especially N-glycosylation, for its full activity. N-Glycoprotein analysis of B. mori fifth instar larvae revealed that high-mannose-type structure was predominant and GlcNAc-linked and fucosylated structures were observed but endogenous galactosyl-, N-acetylgalactosaminyl- and sialyl-N-glycoproteins were undetectable under the standard analytical approach. These results indicate that B. mori genome encodes an α2,6-sialyltransferase, but further investigations of the sialylation potentials are necessary. PMID:26306633

  2. Genomic analysis of carboxyl/cholinesterase genes in the silkworm Bombyx mori

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    Shiotsuki Takahiro

    2010-06-01

    Full Text Available Abstract Background Carboxyl/cholinesterases (CCEs have pivotal roles in dietary detoxification, pheromone or hormone degradation and neurodevelopment. The recent completion of genome projects in various insect species has led to the identification of multiple CCEs with unknown functions. Here, we analyzed the phylogeny, expression and genomic distribution of 69 putative CCEs in the silkworm, Bombyx mori (Lepidoptera: Bombycidae. Results A phylogenetic tree of CCEs in B. mori and other lepidopteran species was constructed. The expression pattern of each B. mori CCE was also investigated by a search of an expressed sequence tag (EST database, and the relationship between phylogeny and expression was analyzed. A large number of B. mori CCEs were identified from a midgut EST library. CCEs expressed in the midgut formed a cluster in the phylogenetic tree that included not only B. mori genes but also those of other lepidopteran species. The silkworm, and possibly also other lepidopteran species, has a large number of CCEs, and this might be a consequence of the large cluster of midgut CCEs. Investigation of intron-exon organization in B. mori CCEs revealed that their positions and splicing site phases were strongly conserved. Several B. mori CCEs, including juvenile hormone esterase, not only showed clustering in the phylogenetic tree but were also closely located on silkworm chromosomes. We investigated the phylogeny and microsynteny of neuroligins in detail, among many CCEs. Interestingly, we found the evolution of this gene appeared not to be conserved between B. mori and other insect orders. Conclusions We analyzed 69 putative CCEs from B. mori. Comparison of these CCEs with other lepidopteran CCEs indicated that they had conserved expression and function in this insect order. The analyses showed that CCEs were unevenly distributed across the genome of B. mori and suggested that neuroligins may have a distinct evolutionary history from other

  3. Mechanism of fluorescent cocoon sex identification for silkworms Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    By using silkworms,Bombyx mori, fluorescent cocoon sex identification (FCSI) as an experimental material, direct fluorescence spectrometry of the cocoon surface indicates that the fluorescent color of silkworm cocoons is made up of two peaks of yellow and blue-purple fluorescence emission. The fluorescent difference between male and female cocoons is attributed to the differential absorption of yellow fluorescent substances by the midgut tissue of 5th instar female silkworms. Thin layer chromatography (TLC) and fluorescent spectra indicate that blue-purple fluorescent substances are composed of at least five blue-purple fluorescent pigments, and yellow fluorescent substances are made up of at least three. UV spectra and AlCl3 color reaction show that the three fluorescent yellow pigments are flavonoids or their glycosides. Silkworm FCSI is due to selective absorption or accumulation of the yellow fluorescent pigments by the posterior midgut cells of female 5th instar larvae. The cells of the FCSI silkworm midgut, especially the cylinder intestinal cells of the posterior midgut have a component which is a yellow fluorescent pigment-specific binding protein that may be vigorously expressed in the 5th instar larvae.

  4. Genome engineering and parthenocloning in the silkworm, Bombyx mori

    Indian Academy of Sciences (India)

    Valeriya Zabelina; Vyacheslav Klymenko; Toshiki Tamura; Karina Doroshenko; Haoyuan Liang; Hideki Sezutsu; František Sehnal

    2015-09-01

    Genetic engineering of the silkworm, Bombyx mori, opens door to the production of new kinds of silk and to the use of silkworms as proteosynthetic bioreactors. The insertion of foreign genes into silkworm genome and the control of their expression by diverse promoters have become possible but are not yet efficient enough for commercial use. Several methods of gene targeting are being developed to minimize position effect on transgene expression and facilitate cloning. Parthenocloning can be exploited to conserve genetic traits and improve selection and amplification of clones containing genes of interest. Some silkworm clones have been bred for decades as genetically stable female stocks whose unfertilized eggs are induced to develop by heat-shock treatment. Any exclusively female generation contains exact copies of the maternal clone-founder genome. Ovaries transplanted in either direction between the standard and the parthenogenetic genotypes yield eggs capable of parthenocloning. In addition, use ofmale larvae as ovary recipients eliminates diapause in eggs produced in the implants. Unfertilized eggs of some silkworm clones respond also to the cold-shock treatment by producing homozygous fertile sons; cloned females can be crossed with their parthenogenetic sons to obtain progeny homozygous for the transgene in both sexes. Rational exploitation of available parthenozygous pools and the use of parthenocloning methods enable rapid fixation and maintenance of the desired genotypes.

  5. Cloning and characterization of nanos gene in silkworm Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    Guoli Zhao; Keping Chen; Qin Yao; Weihua Wang

    2008-01-01

    Gene nanos is a maternal posterior group gene required for normal development of abdominal segments and the germ line in Droso phila. Expression of nanos-related genes is associated with the germ line in a broad variety of other taxa. In this study, the 5'-RACE method and the in silico cloning method are used to isolate the new nanos-like gene of Bombyx mori and the gene obtained is analyzed with bioinformatics tools. The putative protein is expressed in Escherichia coli and the antiserum has been produced in New Zealand white rabbits. The result shows that the nanos cDNA is 1,913 bp in full length and contains a 954 bp open reading frame. The deduced protein has 317 amino acid residues, with a predicted molecular weight of 35 kDa, isoelectric point of 5. 38, and contains a conserved nanos RNA binding domain. The conserved region of the deduced protein shares 73% homology with the nanos protein conserved region of Honeybee (Apis mellifera). This gene has been registered in the GenBank under the accession number EF647589. One encoding se quence of the nanos fragment has been successfully expressed in E. coli. Western blotting analysis indicates that homemade antiserum can specifically detect nanos protein expressed in prokaryotic cells.

  6. Cloning and characterization of nanos gene in silkworm Bombyx mori.

    Science.gov (United States)

    Zhao, Guoli; Chen, Keping; Yao, Qin; Wang, Weihua

    2008-02-01

    Gene nanos is a maternal posterior group gene required for normal development of abdominal segments and the germ line in Drosophila. Expression of nanos-related genes is associated with the germ line in a broad variety of other taxa. In this study, the 5'-RACE method and the in silico cloning method are used to isolate the new nanos-like gene of Bombyx mori and the gene obtained is analyzed with bioinformatics tools. The putative protein is expressed in Escherichia coli and the antiserum has been produced in New Zealand white rabbits. The result shows that the nanos cDNA is 1,913 bp in full length and contains a 954 bp open reading frame. The deduced protein has 317 amino acid residues, with a predicted molecular weight of 35 kDa, isoelectric point of 5. 38, and contains a conserved nanos RNA binding domain. The conserved region of the deduced protein shares 73% homology with the nanos protein conserved region of Honeybee (Apis mellifera). This gene has been registered in the GenBank under the accession number EF647589. One encoding sequence of the nanos fragment has been successfully expressed in E. coli. Western blotting analysis indicates that homemade antiserum can specifically detect nanos protein expressed in prokaryotic cells. PMID:18407054

  7. Vertical transmission of nucleopolyhedrovirus in the silkworm, Bombyx mori L.

    Science.gov (United States)

    Khurad, A M; Mahulikar, A; Rathod, M K; Rai, M M; Kanginakudru, S; Nagaraju, J

    2004-09-01

    Nucleopolyhedrovirus (NPV) was tested for vertical transmission in the silkworm, Bombyx mori. Fifth instar larvae were exposed to four different dosages of BmNPV (830, 1300, 1800, and 2000OBs/larva) and a dosage of about 2000OBs/larva was found suitable for obtaining infected adults. Histopathological studies revealed the infection in susceptible tissues and organs initially, and at later stages of infection cycles the spermatocytes and nurse cells in the young oocytes were infected in the larval rudiments of testis and ovary, respectively. The mating of infected females with uninfected males resulted in significant reduction in fecundity (P < 0.01) and hatching of eggs (P < 0.001) due to transovarial transmission of BmNPV. Mating tests of uninfected females and infected males also confirmed venereal transmission as there was a significant reduction in hatching of eggs (P < 0.01). Further, among the F1 hybrid offspring (infected female x uninfected male) that were infected transovarially, larval progeny died at first and second instar stages, whereas those infected venereally developed acute lethal infection late and died by the end of third and fourth instar stage. PCR amplification and sequencing of 473bp of immediate early-1 (ie-1) gene of BmNPV isolated from the viral-infected parent and the F1 offspring confirmed that the viral infection is vertically transmitted to the progeny. PMID:15491594

  8. Molecular Dissection of Bombyx mori Nucleopolyhedrovirus orf8 Gene

    Institute of Scientific and Technical Information of China (English)

    WonKyung Kang

    2009-01-01

    Viruses including baculoviruses are obligatory parasites, as their genomes do not encode all the proteins required for replication. Therefore, viruses have evolved to exploit the behavior and the physiology of their hosts and often eoevolved with their hosts over millions of years. Recent comparative analyses of complete genome sequences of baculoviruses revealed the patterns of gene acquisitions and losses that have occurred during baculovirus evolution. In addition, knowledge of virus genes has also provided understanding of the mechanism of baculovirus infection including replication, species-specific virulence and host range. The Bm8 gene of Bombyx mori nucleopolyhedrovirus (NPV) and its homologues are found only in group I NPV genomes. The Autographa californica NPV Acl6 gene is a homologue of Bm8 and, encodes a viral structural protein. It has been shown that Bm8/Ac 16 interacts with baculoviral and cellular proteins. Bm8/Ac 16 interacts with baculoviral IE1 that is facilitated by coiled coil domains, and the interaction with IE1 is important for Bin8 function. Ac16 also forms a complex with viral FP25 and cellular actin and associates with membranes via palmitoylation. These data suggested that this gene family encodes a multifunctional protein that accomplishes specific needs of group INPVs.

  9. Hemocytes and hematopoiesis in the silkworm, Bombyx mori

    Directory of Open Access Journals (Sweden)

    F Liu

    2013-10-01

    Full Text Available The silkworm, Bombyx mori, is a typical Lepidopteran insect. In the silkworm hemolymph, there are 5 types of circulating hemocytes that are classified as prohemocytes, granulocytes, plasmatocytes, spherulocytes and oenocytoids. All of them are involved in humoral and cellular immunity either directly or indirectly. Insect hematopoietic organs can produce hemocytes that are continuously released into the circulation. Recent studies indicate that in the hematopoietic organs of silkworm larvae, there are mainly prohemocytes and oenocytoids. Based on in vitro observations, silkworm prohemocytes can differentiate into plasmatocytes and granulocytes, and granulocytes can differentiate into spherulocytes. The silkworm also has a novel type of hematopoiesis. When its hematopoietic organs are extirpated through a surgical operation, circulating hemocytes can still remain at a high level through the wandering stage due to an increase in the level of cell division. Previously, oenocytoids have been considered as the only source of prophenoloxidase (PPO which is an important immunity protein in insects. However, recent studies in different insect species, as well as in the silkworm, show that additional hemocyte types contain PPO. Furthermore, PPO can be produced by epidermal cells in the hindgut of the silkworm. Consequently, the silkworm is a valuable model to study hemocyte development and cellular and humoral immune responses.

  10. Molecular Characterization of Endoplasmic Reticulum Oxidoreductin 1 from Bombyx mori

    Directory of Open Access Journals (Sweden)

    Minchul Seo

    2015-11-01

    Full Text Available We isolated a complementary DNA (cDNA clone encoding endoplasmic reticulum oxidoreductin 1 (bERO1, a specific oxidant of protein disulfide isomerase (PDI from Bombyx mori. This protein has a putative open reading frame (ORF of 489 amino acids and a predicted size of 57.4 kDa. Although bERO1 protein shares less than 57% amino acid sequence homology with other reported ERO1s, it contains two conserved redox active motifs, a Cys-X-X-X-X-Cys motif of N-terminal and Cys-X-X-Cys-X-X-Cys motif of C-terminal. Both motifs are typically present in ERO1 protein family members. The bEro1 mRNA expression was highest in posterior silk gland on the sixth day of the 5th instar larvae. Expression of bEro1 mRNA also markedly increased during endoplasmic reticulum (ER stress induced by stimulation with antimycin, calcium ionophore A23187, dithiothreitol, H2O2, monencin, and tunicamycin. In addition, expression levels of bEro1 exactly coincided with that of bPdi. This is the first result suggesting that bERO1 plays an essential role in ER quality control through the combined activities of bERO1 and bPDI as a catalyst of protein folding in the ER and sustaining cellular redox homeostasis.

  11. Ultrathin Free-Standing Bombyx mori Silk Nanofibril Membranes.

    Science.gov (United States)

    Ling, Shengjie; Jin, Kai; Kaplan, David L; Buehler, Markus J

    2016-06-01

    We report a new ultrathin filtration membrane prepared from silk nanofibrils (SNFs), directly exfoliated from natural Bombyx mori silk fibers to retain structure and physical properties. These membranes can be prepared with a thickness down to 40 nm with a narrow distribution of pore sizes ranging from 8 to 12 nm. Typically, 40 nm thick membranes prepared from SNFs have pure water fluxes of 13 000 L h(-1) m(-2) bar(-1), more than 1000 times higher than most commercial ultrathin filtration membranes and comparable with the highest water flux reported previously. The commercial membranes are commonly prepared from polysulfone, poly(ether sulfone), and polyamide. The SNF-based ultrathin membranes exhibit efficient separation for dyes, proteins, and colloids of nanoparticles with at least a 64% rejection of Rhodamine B. This broad-spectrum filtration membrane would have potential utility in applications such as wastewater treatment, nanotechnology, food industry, and life sciences in part due to the protein-based membrane polymer (silk), combined with the robust mechanical and separation performance features. PMID:27076389

  12. The ionotropic γ-aminobutyric acid receptor gene family of the silkworm, Bombyx mori.

    Science.gov (United States)

    Yu, Lin-Lin; Cui, Ying-Jun; Lang, Guo-Jun; Zhang, Ming-Yan; Zhang, Chuan-Xi

    2010-09-01

    γ-Aminobutyric acid (GABA) is a very important inhibitory neurotransmitter in both vertebrate and invertebrate nervous systems. GABA receptors (GABARs) are known to be the molecular targets of a class of insecticides. Members of the GABAR gene family of the silkworm, Bombyx mori, a model insect of Lepidoptera, have been identified and characterized in this study. All putative silkworm GABAR cDNAs were cloned using the reverse transcriptase polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). Bombyx mori appears to have the largest insect GABAR gene family known to date, including three RDL, one LCCH3, and one GRD subunit. The silkworm RDL1 gene has RNA-editing sites, and the RDL1 and RDL3 genes possess alternative splicing. These mRNA modifications enhance the diversity of the silkworm's GABAR gene family. In addition, truncated transcripts were found for the RDL1 and LCCH3 genes. In particular, the three RDL subunits may have arisen from two duplication events. PMID:20924418

  13. Isozymic variations in specific and nonspecific esterase and its thermostability in silkworm, Bombyx mori L.

    Science.gov (United States)

    Patnaik, Bharat Bhusan; Biswas, Tapati Datta; Nayak, Sandeepta Kumar; Saha, A K; Majumdar, M K

    2012-09-01

    Esterase isozymic variations were documented in the haemolymph of developed multivoltine and bivoltine silkworm breeds during unfavorable seed crop seasons of May - September using á- and â- napthylacetate separately to identify specific and nonspecific esterase having thermotolerant potentiality. Variations existed in the isozyme pattern with three bands (Est-2, 3 and 4) in pure Nistari race and other developed multivoltine and bivoltine breeds. Est-2 and Est-3 were non-specific esterases as they were observed when both á- and â-napthylacetate was used as substrates separately. Est-4 band was observed only with á-napthylacetate as substrate and was therefore confirmed to be specific á-esterase band in the haemolymph of silkworm, Bombyx mori L. Zymograms showed that the non-specific esterase band (Est-3) with R1 of 0.43 and specific á-esterase band (Est-4) with R(f) of 0.32 predominately withstood a temperature of 70 +/- 2 degrees C for a duration of 10 min and were confirmed as thermostable esterases in haemolymph of silkworm, Bombyx mori L. This also categorized the presence of thermostable esterases in developed multivoltine and bivoltine breeds of silkworm, even though the qualitative activity was more in the former than the latter. The qualitative presence of thermostable esterases and their activity could be adopted as an indicative biochemical marker in relation to thermotolerance in silkworm. PMID:23734447

  14. Expression, purification and characterization of methyl DNA binding protein from Bombyx mori.

    Science.gov (United States)

    Uno, Tomohide; Nomura, Yuka; Nakamura, Masahiko; Nakao, Atsushi; Tajima, Shoji; Kanamaru, Kengo; Yamagata, Hiroshi; Iwanaga, Yousuke

    2005-01-01

    A cDNA clone encoding methyl DNA binding domain-containing protein (bMBD2/3) was obtained by homology searches using a Bombyx mori fat body cDNA library. The cDNA encoded a polypeptide with 249 amino acids sharing 54% similarity with the methyl DNA binding protein from Drosophila melanogaster. To characterize the biochemical properties of bMBD2/3, the clone was expressed in Escherichia coli as His-tagged protein. The recombinant protein was purified to homogeneity using Ni-NTA superflow resin and heparin agarose. The protein showed specific methyl DNA binding activity and was phosphorylated by protein kinase in vitro. Immunoblotting using the purified antibody indicated that bMBD2/3 was expressed in almost all tissues. Using west-western blotting analysis, some proteins that interact with bMBD2/3 were identified in the brain. This is the first report that insect MBD is phosphorylated and is present in adult tissues. These results suggest that bMBD2/3 plays important roles in the DNA methylation-specific transcription of Bombyx mori. PMID:16299598

  15. Genome-wide identification and characterization of ATP-binding cassette transporters in the silkworm, Bombyx mori

    OpenAIRE

    Liu, Shumin; Zhou, Shun; Tian, Ling; Guo, Enen; Luan, Yunxia; Zhang, Jianzhen; Li, Sheng

    2011-01-01

    Background The ATP-binding cassette (ABC) transporter superfamily is the largest transporter gene family responsible for transporting specific molecules across lipid membranes in all living organisms. In insects, ABC transporters not only have important functions in molecule transport, but also play roles in insecticide resistance, metabolism and development. Results From the genome of the silkworm, Bombyx mori, we have identified 51 putative ABC genes which are classified into eight subfamil...

  16. Regulation of the innate immune responses in the silkworm, Bombyx mori

    Directory of Open Access Journals (Sweden)

    H Tanaka

    2011-04-01

    Full Text Available Insects possess an effective innate immune system against foreign microorganisms. Innate immunity of insects is divided into two major reaction types: humoral and cellular reactions. Humoral reactions involve soluble proteins in the hemolymph such as phenoloxidase, antimicrobial proteins (AMPs, lysozymes, and lectins, whereas hemocytes mediate cellular reactions such as phagocytosis, encapsulation and nodule formation. In Bombyx mori, six different families of AMPs have been identified: Cecropin, Attacin, Lebocin, Moricin, Gloverin, and Defensin. One lysozyme and three lysozyme-like proteins, one of which is involved in elimination of invading pathogens, are also found in the silkworm. Both lysine-containing peptidoglycan (Lys-PGN and meso-diaminopimelic acid containing peptidoglycan (DAP-PGN trigger expression of AMP genes, probably through the Toll and IMD pathways, respectively. DAP-PGN has stronger elicitor activity than Lys-PGN in B. mori because of the difference in transcriptional activity between BmRelishes and BmRels, which are effectors of the IMD and Toll pathways, respectively. Furthermore, two recognition proteins and a serine protease are involved in activation of prophenoloxidase for melanization, and several C-type lectins, which participated in cellular reactions, were identified in B. mori. Moreover, a paralytic peptide was reported to play important roles in silkworm immunity. Recent development of transgenic technologies and silkworm genome information are expected to accelerate silkworm immunity studies.

  17. Identification of key uric acid synthesis pathway in a unique mutant silkworm Bombyx mori model of Parkinson's disease.

    Directory of Open Access Journals (Sweden)

    Hiroko Tabunoki

    Full Text Available Plasma uric acid (UA levels decrease following clinical progression and stage development of Parkinson's disease (PD. However, the molecular mechanisms underlying decreases in plasma UA levels remain unclear, and the potential to apply mutagenesis to a PD model has not previously been discovered. We identified a unique mutant of the silkworm Bombyx mori (B.mori op. Initially, we investigated the causality of the phenotypic "op" by microarray analysis using our constructed KAIKO functional annotation pipeline. Consequently, we found a novel UA synthesis-modulating pathway, from DJ-1 to xanthine oxidase, and established methods for large-scale analysis of gene expression in B. mori. We found that the mRNA levels of genes in this pathway were significantly lower in B. mori op mutants, indicating that downstream events in the signal transduction cascade might be prevented. Additionally, levels of B.mori tyrosine hydroxylase (TH and DJ-1 mRNA were significantly lower in the brain of B. mori op mutants. UA content was significantly lower in the B. mori op mutant tissues and hemolymph. The possibility that the B. mori op mutant might be due to loss of DJ-1 function was supported by the observed vulnerability to oxidative stress. These results suggest that UA synthesis, transport, elimination and accumulation are decreased by environmental oxidative stress in the B. mori op mutant. In the case of B. mori op mutants, the relatively low availability of UA appears to be due both to the oxidation of DJ-1 and to its expenditure to mitigate the effects of environmental oxidative stress. Our findings are expected to provide information needed to elucidate the molecular mechanism of decreased plasma UA levels in the clinical stage progression of PD.

  18. Expansion of CRISPR targeting sites in Bombyx mori.

    Science.gov (United States)

    Zeng, Baosheng; Zhan, Shuai; Wang, Yueqiang; Huang, Yuping; Xu, Jun; Liu, Qun; Li, Zhiqian; Huang, Yongping; Tan, Anjiang

    2016-05-01

    The CRISPR/Cas9 system has been proven as a revolutionary genome engineering tool. In most cases, single guide RNA (sgRNA) targeting sites have been designed as GN19NGG or GGN18NGG, because of restriction of the initiation nucleotide for RNA Pol III promoters. Here, we demonstrate that the U6 promoter from a lepidopteran model insect, Bombyx mori, effectively expressed the sgRNA initiated with any nucleotide bases (adenine, thymine, guanine or cytosine), which further expands the CRISPR targeting space. A detailed expansion index in the genome was analysed when N20NGG was set as the CRISPR targeting site instead of GN19NGG, and revealed a significant increase of suitable targets, with the highest increase occurring on the Z sex chromosome. Transfection of different types of N20NGG sgRNAs targeting the enhanced green fluorescent protein (EGFP) combined with Cas9, significantly reduced EGFP expression in the BmN cells. An endogenous gene, BmBLOS2, was also disrupted by using various types of N20NGG sgRNAs, and the cleavage efficiency of N20NGG sgRNAs with different initial nucleotides and GC contents was evaluated in vitro. Furthermore, transgenic silkworms expressing Cas9 and sgRNAs targeting the BmBLOS2 gene were generated with many types of mutagenesis. The typical transparent skin phenotype in knock-out silkworms was stable and inheritable, suggesting that N20NGG sgRNAs function sufficiently in vivo. Our findings represent a renewal of CRISPR/Cas9 target design and will greatly facilitate insect functional genetics research. PMID:27032928

  19. Electroporation, an alternative to biolistics for transfection of Bombyx mori embryos and larval tissues

    OpenAIRE

    Jean-Luc Thomas

    2003-01-01

    There are few powerful techniques available to transfect insect tissues. We previously used biolistics to transfect Bombyx mori embryos, and larval and pupal tissues (Thomas J-L et al. 2001. Journal of Insect Science 1/9, Kravariti L et al. 2001. Insect Biochemistry and Molecular Biology 31: 473–479). As the main limitation was the irregularity in results we explored electroporation as an alternative technique by adapting techniques used for chicken embryos to B. mori embryos. By injecting th...

  20. Comparative proteome analysis of multi-layer cocoon of the silkworm, Bombyx mori.

    Science.gov (United States)

    Zhang, Yan; Zhao, Ping; Dong, Zhaoming; Wang, Dandan; Guo, Pengchao; Guo, Xiaomeng; Song, Qianru; Zhang, Weiwei; Xia, Qingyou

    2015-01-01

    Bombyx mori cocoon has a multi-layer structure that provides optimal protection for silkworm pupa. Research on the mechanical properties of the multi-layer structure revealed structure-property relationships of the cocoon. Here, we investigated the protein components of the B. mori cocoon in terms of its multi-layer structure. Liquid chromatography-tandem mass spectrometry identified 286 proteins from the multiple cocoon layers. In addition to fibroins and sericins, we identified abundant protease inhibitors, seroins and proteins of unknown function. By comparing protein abundance across layers, we found that the outermost layer contained more sericin1 and protease inhibitors and the innermost layer had more seroin1. As many as 36 protease inhibitors were identified in cocoons, showing efficient inhibitory activities against a fungal protease. Thus, we propose that more abundant protease inhibitors in the outer cocoon layers may provide better protection for the cocoon. This study increases our understanding of the multi-layer mechanism of cocoons, and helps clarify the biological characteristics of cocoons. The data have been deposited to the ProteomeXchange with identifier PXD001469. PMID:25860555

  1. Analysis of tissue-specific region in sericin 1 gene promoter of Bombyx mori

    International Nuclear Information System (INIS)

    The gene encoding sericin 1 (Ser1) of silkworm (Bombyx mori) is specifically expressed in the middle silk gland cells. To identify element involved in this transcription-dependent spatial restriction, truncation of the 5' terminal from the sericin 1 (Ser1) promoter is studied in vivo. A 209 bp DNA sequence upstream of the transcriptional start site (-586 to -378) is found to be responsible for promoting tissue-specific transcription. Analysis of this 209 bp region by overlapping deletion studies showed that a 25 bp region (-500 to -476) suppresses the ectopic expression of the Ser1 promoter. An unknown factor abundant in fat body nuclear extracts is shown to bind to this 25 bp fragment. These results suggest that this 25 bp region and the unknown factor are necessary for determining the tissue-specificity of the Ser1 promoter

  2. Silkworm (Bombyx mori) hemocytes do not produce reactive oxygen metabolites as a part of defense mechanisms

    Czech Academy of Sciences Publication Activity Database

    Hyršl, P.; Číž, Milan; Kubala, Lukáš; Lojek, Antonín

    2004-01-01

    Roč. 49, č. 3 (2004), s. 315-319. ISSN 0015-5632 R&D Projects: GA AV ČR IBS5004009 Institutional research plan: CEZ:AV0Z5004920 Keywords : hemocytes * Bombyx mori * reactive oxygen species Subject RIV: BO - Biophysics Impact factor: 1.034, year: 2004

  3. Cloning and expression of a portion of \\kur{Bombyx Mori Ser/2} gene

    OpenAIRE

    STAŠKOVÁ, Tereza

    2009-01-01

    A 897 bp section of Bombyx mori Ser-2 gene was amplified, cloned into a bacterial expression vector, and used to prepare a recombinant sericin-like protein. The protein is expected to contain domain supporting growth of mammalian cells. It will be used in assays verifying this assumption; if confirmed, recombinant protein will be considered for preparations of scaffold guiding tissue reconstruction.

  4. Effect of pyridoxine on the reproduction of the mulberry silkworm, Bombyx mori L.

    Directory of Open Access Journals (Sweden)

    SI Faruki

    2005-03-01

    Full Text Available The present investigation reports the effects of vitamin B6, also known as pyridoxine supplementedfeed on the reproductive potential of Bombyx mori L. All the concentrations of the vitamin significantlyreduced the fecundity. Egg-viability was also reduced at all the concentrations, but the differences werenot significant in comparison to control.

  5. Comparative proteome analysis of silkworm in its susceptibility and resistance responses to Bombyx mori densonucleosis virus.

    Science.gov (United States)

    Chen, Hui-Qing; Yao, Qin; Bao, Fang; Chen, Ke-Ping; Liu, Xiao-Yong; Li, Jun; Wang, Lin

    2012-01-01

    Bombyx mori densonucleosis virus (BmDNV) is one of the most disastrous viruses in cocoon production. Silkworm resistance to BmDNV has been examined previously using a number of traditional biochemical and molecular techniques. In this study, a near isogenic line, BC(6), was constructed to eliminate the difference in inherited background, which has 99.9% identity with the susceptible strain but carries a resistant gene. We utilized a proteomic approach involving two-dimensional differential gel electrophoresis and mass spectrometry to examine changes in the midgut proteins from the susceptible and resistant silkworm larvae infected with BmDNV. The protein profiles were compared and 9 differentially expressed proteins were identified by mass spectrometry. In the resistant strains, the heat-shock 70-kDa protein cognate, cytochrome P450, vacuolar ATP synthase subunit B, arginine kinase, vacuolar ATP synthase subunit D and glutathione S-transferase sigma were strongly upregulated and α-tubulin was downregulated. Our results imply that these upregulated genes and the downregulated genes might be involved in B. mori immune responses against BmDNV-Z infection. PMID:21242662

  6. Mechanisms of nodule-specific melanization in the hemocoel of the silkworm, Bombyx mori.

    Science.gov (United States)

    Shu, Min; Mang, Dingze; Fu, Gege Sun; Tanaka, Shiho; Endo, Haruka; Kikuta, Shingo; Sato, Ryoichi

    2016-03-01

    In the insect immune system, nodules are known to be a product of the cellular response against microorganisms and may be a preferential target for melanization. However, the mechanism of nodule-preferential melanization remains to be explored. In this study, we identified several mechanisms of nodule-preferential melanization by analyzing congregation and the activation of several factors involved in the prophenoloxidase (proPO)-activating system in the silkworm, Bombyx mori. Microorganism-binding assays revealed that B. mori larval plasma have an effective invading microorganism-surveillance network consisting of at least six pattern-recognition receptors (PRRs). We also found that a hemolymph serine proteinase, BmHP14, can bind to Saccharomyces cerevisiae. Pull-down assays showed that PRR C-type lectins form protein complexes with serine proteinase homologs, BmSPH1 and BmSPH2, which leads to the activated forms of BmSPH1 and BmSPH2 being gathered on microorganisms and trapped in nodules. Immunostaining analysis revealed that most factors in the proPO-activating system and some factors in the triggering system for antimicrobial peptide production exist in the granules of hemocytes which can gather in nodules. Western blot analysis showed that factors in the proPO-activating system are congregated in formed nodules by their concentration in plasma and aggregating hemocytes. PMID:26707571

  7. Silvernanotherapy on the viral borne disease of silkworm Bombyx mori L

    International Nuclear Information System (INIS)

    Antiviral assays of chemically and biologically synthesized silver nanoparticles were made against BmNPV (Bombyx mori Nuclear Polyhedrosis Virus). Reduction of silver ions by sodium citrate and Spirulina platensis led to the formation of spherical silver nanoparticles of 40–60 and 7–16 nm size. Single cell protein (Spirulina platensis)-synthesized silver nanoparticles showed the strongest antiviral activity. Immunological studies made on the silkworm Bombyx mori disclosed that a significant increase in the total hemocyte count and differential hemocyte count due to S. platensis-synthesized silver nanoparticles supplementation. Improvement in the defense mechanism was noticed from the strengthened peritrophic membrane of the digestive tract and the increased total protein. Overall, the results presented illustrate that single cell protein-synthesized silver nanoparticles supplementation is effective in controlling viral-borne diseases of the silkworm.

  8. Mapping and recombination analysis of two moth colour mutations, Black moth and Wild wing spot, in the silkworm Bombyx mori.

    Science.gov (United States)

    Ito, K; Katsuma, S; Kuwazaki, S; Jouraku, A; Fujimoto, T; Sahara, K; Yasukochi, Y; Yamamoto, K; Tabunoki, H; Yokoyama, T; Kadono-Okuda, K; Shimada, T

    2016-01-01

    Many lepidopteran insects exhibit body colour variations, where the high phenotypic diversity observed in the wings and bodies of adults provides opportunities for studying adaptive morphological evolution. In the silkworm Bombyx mori, two genes responsible for moth colour mutation, Bm and Ws, have been mapped to 0.0 and 14.7 cM of the B. mori genetic linkage group 17; however, these genes have not been identified at the molecular level. We performed positional cloning of both genes to elucidate the molecular mechanisms that underlie the moth wing- and body-colour patterns in B. mori. We successfully narrowed down Bm and Ws to ~2-Mb-long and 100-kb-long regions on the same scaffold Bm_scaf33. Gene prediction analysis of this region identified 77 candidate genes in the Bm region, whereas there were no candidate genes in the Ws region. Fluorescence in-situ hybridisation analysis in Bm mutant detected chromosome inversion, which explains why there are no recombination in the corresponding region. The comparative genomic analysis demonstrated that the candidate regions of both genes shared synteny with a region associated with wing- and body-colour variations in other lepidopteran species including Biston betularia and Heliconius butterflies. These results suggest that the genes responsible for wing and body colour in B. mori may be associated with similar genes in other Lepidoptera. PMID:26219230

  9. Carbonic anhydrase generates a pH gradient in Bombyx mori silk glands.

    Science.gov (United States)

    Domigan, L J; Andersson, M; Alberti, K A; Chesler, M; Xu, Q; Johansson, J; Rising, A; Kaplan, D L

    2015-10-01

    Silk is a protein of interest to both biological and industrial sciences. The silkworm, Bombyx mori, forms this protein into strong threads starting from soluble silk proteins using a number of biochemical and physical cues to allow the transition from liquid to fibrous silk. A pH gradient has been measured along the gland, but the methodology employed was not able to precisely determine the pH at specific regions of interest in the silk gland. Furthermore, the physiological mechanisms responsible for the generation of this pH gradient are unknown. In this study, concentric ion selective microelectrodes were used to determine the luminal pH of B. mori silk glands. A gradient from pH 8.2 to 7.2 was measured in the posterior silk gland, with a pH 7 throughout the middle silk gland, and a gradient from pH 6.8 to 6.2 in the beginning of the anterior silk gland where silk processing into fibers occurs. The small diameter of the most anterior region of the anterior silk gland prevented microelectrode access in this region. Using a histochemical method, the presence of active carbonic anhydrase was identified in the funnel and anterior silk gland of fifth instar larvae. The observed pH gradient collapsed upon addition of the carbonic anhydrase inhibitor methazolamide, confirming an essential role for this enzyme in pH regulation in the B. mori silk gland. Plastic embedding of whole silk glands allowed clear visualization of the morphology, including the identification of four distinct epithelial cell types in the gland and allowed correlations between silk gland morphology and silk stages of assembly related to the pH gradient. B. mori silk glands have four different epithelial cell types, one of which produces carbonic anhydrase. Carbonic anhydrase is necessary for the mechanism that generates an intraluminal pH gradient, which likely regulates the assembly of silk proteins and then the formation of fibers from soluble silk proteins. These new insights into native silk

  10. Existence of Prophenoloxidase in Wing Discs: A Source of Plasma Prophenoloxidase in the Silkworm, Bombyx mori

    OpenAIRE

    Diao, Yupu; Lu, Anrui; Yang, Bing; Hu, Wenli; Peng, Qing; Ling, Qing-Zhi; Beerntsen, Brenda T.; Söderhäll, Kenneth; Ling, Erjun

    2012-01-01

    In insects, hemocytes are considered as the only source of plasma prophenoloxidase (PPO). PPO also exists in the hemocytes of the hematopoietic organ that is connected to the wing disc of Bombyx mori. It is unknown whether there are other cells or tissues that can produce PPO and release it into the hemolymph besides circulating hemocytes. In this study, we use the silkworm as a model to explore this possibility. Through tissue staining and biochemical assays, we found that wing discs contain...

  11. Genetic Divergence, Implication of Diversity, and Conservation of Silkworm, Bombyx mori

    OpenAIRE

    Bindroo, Bharat Bhusan; Manthira Moorthy, Shunmugam

    2014-01-01

    Genetic diversity is critical to success in any crop breeding and it provides information about the quantum of genetic divergence and serves a platform for specific breeding objectives. It is one of the three forms of biodiversity recognized by the World Conservation Union (IUCN) as deserving conservation. Silkworm Bombyx mori, an economically important insect, reported to be domesticated over 5000 years ago by human to meet his requirements. Genetic diversity is a particular concern because ...

  12. Enhancing Effect of Glycerol on the Tensile Properties of Bombyx mori Cocoon Sericin Films

    OpenAIRE

    Liangjun Zhu; Lei Yang; Sijia Min; Haiping Zhang; Lianxia Deng; Mingying Yang

    2011-01-01

    An environmental physical method described herein was developed to improve the tensile properties of Bombyx mori cocoon sericin films, by using the plasticizer of glycerol, which has a nontoxic effect compared with other chemical crosslinkers. The changes in the tensile characteristics and the structure of glycerolated (0–40 wt% of glycerol) sericin films were investigated. Sericin films, both in dry and wet states, showed enhanced tensile properties, which might be regulated by the addition ...

  13. Effect of Venom from the Jellyfish Nemopilema nomurai on the Silkworm Bombyx mori L.

    Directory of Open Access Journals (Sweden)

    Huahua Yu

    2015-09-01

    Full Text Available The silkworm Bombyx mori L. (B. mori has a significant impact on the economy by producing more than 80% of the globally produced raw silk. The exposure of silkworm to pesticides may cause adverse effects on B. mori, such as a reduction in the production and quality of silk. This study aims to assay the effect of venom from the jellyfish Nemopilema nomurai on growth, cuticle and acetylcholinesterase (AChE activity of the silkworm B. mori by the leaf dipping method. The experimental results revealed that the four samples caused neither antifeeding nor a lethal effect on B. mori. The sample SFV inhibited B. mori growth after 6 days of treatment in a dose-dependent manner. The samples SFV, DSFV and Fr-1 inhibited the precipitation and synthesis of chitin in the cuticle after 12 and 14 days of treatment. In the case of the four samples, the AChE was significantly improved after 14 days of treatment.

  14. Advanced technologies for genetically manipulating the silkworm Bombyx mori, a model Lepidopteran insect

    Science.gov (United States)

    Xu, Hanfu; O'Brochta, David A.

    2015-01-01

    Genetic technologies based on transposon-mediated transgenesis along with several recently developed genome-editing technologies have become the preferred methods of choice for genetically manipulating many organisms. The silkworm, Bombyx mori, is a Lepidopteran insect of great economic importance because of its use in silk production and because it is a valuable model insect that has greatly enhanced our understanding of the biology of insects, including many agricultural pests. In the past 10 years, great advances have been achieved in the development of genetic technologies in B. mori, including transposon-based technologies that rely on piggyBac-mediated transgenesis and genome-editing technologies that rely on protein- or RNA-guided modification of chromosomes. The successful development and application of these technologies has not only facilitated a better understanding of B. mori and its use as a silk production system, but also provided valuable experiences that have contributed to the development of similar technologies in non-model insects. This review summarizes the technologies currently available for use in B. mori, their application to the study of gene function and their use in genetically modifying B. mori for biotechnology applications. The challenges, solutions and future prospects associated with the development and application of genetic technologies in B. mori are also discussed. PMID:26108630

  15. Identification of candidate aldehyde oxidases from the silkworm Bombyx mori potentially involved in antennal pheromone degradation.

    Science.gov (United States)

    Pelletier, Julien; Bozzolan, Françoise; Solvar, Marthe; François, Marie-Christine; Jacquin-Joly, Emmanuelle; Maïbèche-Coisne, Martine

    2007-12-01

    Signal inactivation is a crucial step in the dynamic of olfactory process and involves various Odorant-Degrading Enzymes. In the silkworm Bombyx mori, one of the best models for studying olfaction in insects, the involvement of an antennal-specific aldehyde oxidase in the degradation of the sex pheromone component bombykal has been demonstrated over the three past decades by biochemical studies. However, the corresponding enzyme has never been characterized at the molecular level. Bioinformatic screening of B. mori genome and molecular approaches have been used to isolate several candidate sequences of aldehyde oxidases. Two interesting antennal-expressed genes have been further characterized and their putative functions are discussed in regard to their respective expression pattern and to our knowledge on aldehyde oxidase properties. Interestingly, one gene appeared as specifically expressed in the antennae of B. mori and associated in males with the bombykal-sensitive sensilla, strongly suggesting that it could encode for the previously biochemically characterized enzyme. PMID:17904312

  16. Interactions between fibroin and sericin proteins from Antheraea pernyi and Bombyx mori silk fibers.

    Science.gov (United States)

    Du, Shan; Zhang, Jin; Zhou, Wei T; Li, Quan X; Greene, George W; Zhu, Hai J; Li, Jing L; Wang, Xun G

    2016-09-15

    Silkworm silk fibers are core-shell composites of fibroin and sericin proteins. Studying the interactions between fibroin and sericin is essential for understanding the properties of these composites. It is observed that compared to the domestic silk cocoon Bombyx mori (B. mori), the adhesion between fibroin and sericin from the wild silk cocoon, Antheraea pernyi (A. pernyi), is significantly stronger with a higher degree of heterogeneity. The adsorption of A. pernyi sericin on its fibroin is almost twice the value for B. mori sericin on fibroin, both showing a monolayer Langmuir adsorption. (1)H NMR and FTIR studies demonstrate on a molecular level the stronger interactions and the more intensive complex formation between A. pernyi fibroin and sericin, facilitated by the hydrogen bonding between glycine and serine. The findings of this study may help the design of composites with superior interfacial adhesion between different components. PMID:27314644

  17. Precocious metamorphosis in the juvenile hormone-deficient mutant of the silkworm, Bombyx mori.

    Directory of Open Access Journals (Sweden)

    Takaaki Daimon

    Full Text Available Insect molting and metamorphosis are intricately governed by two hormones, ecdysteroids and juvenile hormones (JHs. JHs prevent precocious metamorphosis and allow the larva to undergo multiple rounds of molting until it attains the proper size for metamorphosis. In the silkworm, Bombyx mori, several "moltinism" mutations have been identified that exhibit variations in the number of larval molts; however, none of them have been characterized molecularly. Here we report the identification and characterization of the gene responsible for the dimolting (mod mutant that undergoes precocious metamorphosis with fewer larval-larval molts. We show that the mod mutation results in complete loss of JHs in the larval hemolymph and that the mutant phenotype can be rescued by topical application of a JH analog. We performed positional cloning of mod and found a null mutation in the cytochrome P450 gene CYP15C1 in the mod allele. We also demonstrated that CYP15C1 is specifically expressed in the corpus allatum, an endocrine organ that synthesizes and secretes JHs. Furthermore, a biochemical experiment showed that CYP15C1 epoxidizes farnesoic acid to JH acid in a highly stereospecific manner. Precocious metamorphosis of mod larvae was rescued when the wild-type allele of CYP15C1 was expressed in transgenic mod larvae using the GAL4/UAS system. Our data therefore reveal that CYP15C1 is the gene responsible for the mod mutation and is essential for JH biosynthesis. Remarkably, precocious larval-pupal transition in mod larvae does not occur in the first or second instar, suggesting that authentic epoxidized JHs are not essential in very young larvae of B. mori. Our identification of a JH-deficient mutant in this model insect will lead to a greater understanding of the molecular basis of the hormonal control of development and metamorphosis.

  18. Precocious metamorphosis in the juvenile hormone-deficient mutant of the silkworm, Bombyx mori.

    Science.gov (United States)

    Daimon, Takaaki; Kozaki, Toshinori; Niwa, Ryusuke; Kobayashi, Isao; Furuta, Kenjiro; Namiki, Toshiki; Uchino, Keiro; Banno, Yutaka; Katsuma, Susumu; Tamura, Toshiki; Mita, Kazuei; Sezutsu, Hideki; Nakayama, Masayoshi; Itoyama, Kyo; Shimada, Toru; Shinoda, Tetsuro

    2012-01-01

    Insect molting and metamorphosis are intricately governed by two hormones, ecdysteroids and juvenile hormones (JHs). JHs prevent precocious metamorphosis and allow the larva to undergo multiple rounds of molting until it attains the proper size for metamorphosis. In the silkworm, Bombyx mori, several "moltinism" mutations have been identified that exhibit variations in the number of larval molts; however, none of them have been characterized molecularly. Here we report the identification and characterization of the gene responsible for the dimolting (mod) mutant that undergoes precocious metamorphosis with fewer larval-larval molts. We show that the mod mutation results in complete loss of JHs in the larval hemolymph and that the mutant phenotype can be rescued by topical application of a JH analog. We performed positional cloning of mod and found a null mutation in the cytochrome P450 gene CYP15C1 in the mod allele. We also demonstrated that CYP15C1 is specifically expressed in the corpus allatum, an endocrine organ that synthesizes and secretes JHs. Furthermore, a biochemical experiment showed that CYP15C1 epoxidizes farnesoic acid to JH acid in a highly stereospecific manner. Precocious metamorphosis of mod larvae was rescued when the wild-type allele of CYP15C1 was expressed in transgenic mod larvae using the GAL4/UAS system. Our data therefore reveal that CYP15C1 is the gene responsible for the mod mutation and is essential for JH biosynthesis. Remarkably, precocious larval-pupal transition in mod larvae does not occur in the first or second instar, suggesting that authentic epoxidized JHs are not essential in very young larvae of B. mori. Our identification of a JH-deficient mutant in this model insect will lead to a greater understanding of the molecular basis of the hormonal control of development and metamorphosis. PMID:22412378

  19. A Hox Gene, Antennapedia, Regulates Expression of Multiple Major Silk Protein Genes in the Silkworm Bombyx mori.

    Science.gov (United States)

    Tsubota, Takuya; Tomita, Shuichiro; Uchino, Keiro; Kimoto, Mai; Takiya, Shigeharu; Kajiwara, Hideyuki; Yamazaki, Toshimasa; Sezutsu, Hideki

    2016-03-25

    Hoxgenes play a pivotal role in the determination of anteroposterior axis specificity during bilaterian animal development. They do so by acting as a master control and regulating the expression of genes important for development. Recently, however, we showed that Hoxgenes can also function in terminally differentiated tissue of the lepidopteranBombyx mori In this species,Antennapedia(Antp) regulates expression of sericin-1, a major silk protein gene, in the silk gland. Here, we investigated whether Antpcan regulate expression of multiple genes in this tissue. By means of proteomic, RT-PCR, and in situ hybridization analyses, we demonstrate that misexpression of Antpin the posterior silk gland induced ectopic expression of major silk protein genes such assericin-3,fhxh4, and fhxh5 These genes are normally expressed specifically in the middle silk gland as is Antp Therefore, the evidence strongly suggests that Antpactivates these silk protein genes in the middle silk gland. The putativesericin-1 activator complex (middle silk gland-intermolt-specific complex) can bind to the upstream regions of these genes, suggesting that Antpdirectly activates their expression. We also found that the pattern of gene expression was well conserved between B. moriand the wild species Bombyx mandarina, indicating that the gene regulation mechanism identified here is an evolutionarily conserved mechanism and not an artifact of the domestication of B. mori We suggest that Hoxgenes have a role as a master control in terminally differentiated tissues, possibly acting as a primary regulator for a range of physiological processes. PMID:26814126

  20. Expression of RYamide in the nervous and endocrine system of Bombyx mori.

    Science.gov (United States)

    Roller, Ladislav; Čižmár, Daniel; Bednár, Branislav; Žitňan, Dušan

    2016-06-01

    RYamides are neuropeptides encoded by a gene whose precise expression and function have not yet been determined. We identified the RYamide gene transcript (fmgV1g15f, SilkBase database) and predicted two candidates for G-protein coupled RYamide receptors (A19-BAG68418 and A22-BAG68421) in the silkworm Bombyx mori. We cloned the RYamide transcript and described its spatial expression using in situ hybridisation. In the larval central nervous system (CNS) expression of RYamide was restricted to 12-14 small neurons in the brain and two posterior neurons in the terminal abdominal ganglion. During metamorphosis their number decreased to eight protocerebral neurons in the adults. Multiple staining, using various insect neuropeptide antibodies, revealed that neurons expressing RYamide are different from other peptidergic cells in the CNS. We also found RYamide expression in the enteroendocrine cells (EC) of the anterior midgut of larvae, pupae and adults. Two minor subpopulations of these EC were also immunoreactive to antibodies against tachykinin and myosupressin. This expression pattern suggests RYamides may play a role in the regulation of feeding and digestion. PMID:26896568

  1. Possible effect of 30K proteins in embryonic development of silkworm Bombyx mori.

    Science.gov (United States)

    Zhong, Bo-Xiong; Li, Jian-Ke; Lin, Jian-Rong; Liang, Jian-She; Su, Song-Kun; Xu, Hai-Sheng; Yan, Hai-Yan; Zhang, Ping-Bo; Fujii, Hiroshi

    2005-05-01

    The silkworm Bombyx mori possesses a 30K protein family of 3x10(4) Da, the biological functions of which have not been fully identified. The relationship between the 30K protein family and the embryonic development of temperature sensitive sex-linked mutant strain of silkworm was investigated by two dimensional polyacrylamide gel electrophoresis (2D-PAGE) and Matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). The results show that protein spots 1-5 of the 30K protein family, mainly existing in normal strain, are possibly related to embryonic development. The early consumption of a 30K protein named 6G1-30K-1 and the accumulation of 30K proteins named 6G1-30K-3 and 6G1-30K-4 are likely caused by the destruction of physiological balance in normal embryonic development, which may lead to lower hatchability of the temperature sensitive strain. The results suggest that reasonable metabolism of 30K proteins is a prerequisite for the embryo's normal development. PMID:15880265

  2. Combinatory annotation of cell membrane receptors and signalling pathways of Bombyx mori prothoracic glands

    Science.gov (United States)

    Moulos, Panagiotis; Samiotaki, Martina; Panayotou, George; Dedos, Skarlatos G.

    2016-01-01

    The cells of prothoracic glands (PG) are the main site of synthesis and secretion of ecdysteroids, the biochemical products of cholesterol conversion to steroids that shape the morphogenic development of insects. Despite the availability of genome sequences from several insect species and the extensive knowledge of certain signalling pathways that underpin ecdysteroidogenesis, the spectrum of signalling molecules and ecdysteroidogenic cascades is still not fully comprehensive. To fill this gap and obtain the complete list of cell membrane receptors expressed in PG cells, we used combinatory bioinformatic, proteomic and transcriptomic analysis and quantitative PCR to annotate and determine the expression profiles of genes identified as putative cell membrane receptors of the model insect species, Bombyx mori, and subsequently enrich the repertoire of signalling pathways that are present in its PG cells. The genome annotation dataset we report here highlights modules and pathways that may be directly involved in ecdysteroidogenesis and aims to disseminate data and assist other researchers in the discovery of the role of such receptors and their ligands. PMID:27576083

  3. Possible Effect of 30K Proteins in Embryonic Development of Silkworm Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    Bo-Xiong ZHONG; Jian-Ke LI; Jian-Rong LIN; Jian-She LIANG; Song-Kun SU; Hai-Sheng XU; Hai-Yan YAN; Ping-Bo ZHANG; Hiroshi FUJII

    2005-01-01

    The silkworm Bombyx mori possesses a 30K protein family of 3×104 Da, the biological functions of which have not been fully identified. The relationship between the 30K protein family and the embryonic development of temperature sensitive sex-linked mutant strain of silkworm was investigated by two dimensional polyacrylamide gel electrophoresis (2D-PAGE) and Matrix assisted laser desorption ionizationtime of flight mass spectrometry (MALDI-TOF MS). The results show that protein spots 1-5 of the 30K protein family, mainly existing in normal strain, are possibly related to embryonic development. The early consumption of a 30K protein named 6G1-30K-1 and the accumulation of 30K proteins named 6G1-30K-3and 6G 1-30K-4 are likely caused by the destruction of physiological balance in normal embryonic development,which may lead to lower hatchability of the temperature sensitive strain. The results suggest that reasonable metabolism of 30K proteins is a prerequisite for the embryo's normal development.

  4. Firing activity of "diapause hormone" producing cells in the male silkmoth, Bombyx mori.

    Science.gov (United States)

    Ichikawa, Toshio; Suenobu, Akiko

    2003-08-01

    Diapause hormone (DH) originally identified to be a factor originating from neurosecretory cells in the suboesophageal ganglion acts on developing ovaries to produce diapause eggs in a female silkmoth, Bombyx mori. A male silkmoth has homologous neurosecretory cells, but little is known of the physiological nature of the cells and actions of their products. We examined the long-term firing activity of putative DH-producing neurosecretory cells and hormonal activity of their products in male pupae that had been experienced different environmental regimens for diapause induction. Firing activity patterns of male labial cells strongly depended on diapause types of pupae: cells in a diapause-type male were active throughout the pupal period, whereas the same cells in a non-diapause-type male were usually inactive during the early two-thirds of the pupal period. A male pupa with electrically active labial cells could induce diapause eggs in a female pupa connected parabiotically to that male. The firing activity of male neurosecretory cells and hormonal action of their products are qualitatively the same as in the female previously examined. We suggest that there is no evident sexual dimorphism in the physiological and biochemical nature of neurosecretory cells producing DH and the amidated peptide DH has different functions in a male. PMID:12951400

  5. Microarray analysis of New Green Cocoon associated genes in silkworm, Bombyx mori.

    Science.gov (United States)

    Lu, Ya-Ru; He, Song-Zhen; Tong, Xiao-Ling; Han, Min-Jin; Li, Chun-Lin; Li, Zhi-Quan; Dai, Fang-Yin

    2016-06-01

    Green cocoons in silkworm, Bombyx mori, are caused by flavonoids accumulation in the silk proteins, fibroin and sericin. Despite the economic value of natural green cocoon and medical value of flavonoids, there is limited understanding of the molecular mechanism regulating flavonoids uptake in silkworm, which is tightly associated with the trait of green cocoon. The purpose of this study is to perform a comprehensive analysis to understand the molecular mechanisms of flavonoids uptake in silkworm based on microarray analyses. The study subject was the New Green Cocoon from the silkworm strains, G200 and N100, a new spontaneous dominant green cocoon trait identified in the 2000s. The genes regulating this trait are independent of other green cocoon genes previously reported. Genome-wide gene expression was compared between the New Green Cocoon producing silkworm strains, G200 and N100, and the control sample, which is the white cocoon producing strain 872B. Among these strains, N100 and 872B are near-isogenic lines. The results showed that 130 genes have consistently changing expression patterns in the green cocoon strains when compared with the white cocoon strain. Among these, we focused on the genes related to flavonoids metabolism and absorption, such as sugar transporter genes and UDP-glucosyltransferase genes. Based on our findings, we propose the potential mechanisms for flavonoids absorption and metabolism in silkworm. Our results imply that silkworm might be used as an underlying model for flavonoids in pharmaceutical research. PMID:26936509

  6. Analysis of proteome dynamics inside the silk gland lumen of Bombyx mori.

    Science.gov (United States)

    Dong, Zhaoming; Zhao, Ping; Zhang, Yan; Song, Qianru; Zhang, Xiaolu; Guo, Pengchao; Wang, Dandan; Xia, Qingyou

    2016-01-01

    The silk gland is the only organ where silk proteins are synthesized and secreted in the silkworm, Bombyx mori. Silk proteins are stored in the lumen of the silk gland for around eight days during the fifth instar. Determining their dynamic changes is helpful for clarifying the secretion mechanism of silk proteins. Here, we identified the proteome in the silk gland lumen using liquid chromatography-tandem mass spectrometry, and demonstrated its changes during two key stages. From day 5 of the fifth instar to day 1 of wandering, the abundances of fibroins, sericins, seroins, and proteins of unknown functions increased significantly in different compartments of the silk gland lumen. As a result, these accumulated proteins constituted the major cocoon components. In contrast, the abundances of enzymes and extracellular matrix proteins decreased in the silk gland lumen, suggesting that they were not the structural constituents of silk. Twenty-five enzymes may be involved in the regulation of hormone metabolism for proper silk gland function. In addition, the metabolism of other non-proteinous components such as chitin and pigment were also discussed in this study. PMID:27102218

  7. Serpin-15 from Bombyx mori inhibits prophenoloxidase activation and expression of antimicrobial peptides.

    Science.gov (United States)

    Liu, Dongran; Wang, Lei; Yang, Liu; Qian, Cen; Wei, Guoqing; Dai, Lishang; Li, Jun; Zhu, Baojian; Liu, Chaoliang

    2015-07-01

    Serine protease inhibitors (SPIs) play a key role in physiological responses by controlling protease activities. In this study, we studied the biochemical functions of serpin-15, an SPI, from Bombyx mori (Bmserpin-15). Recombinant Bmserpin-15 was expressed in Escherichia coli cells and used to raise rabbit anti-Bmserpin-15 polyclonal antibodies. Bmserpin-15 mRNA and protein expression was detected in all tested tissues, particularly in the fat body and silk gland. After challenge with four different microorganisms (Escherichia coli, Beauveria bassiana, Micrococcus luteus and B. mori nuclear polyhedrosis virus), the expressions of Bmserpin-15 mRNA and protein were induced significantly, particularly by B. bassiana and M. luteus. Recombinant Bmserpin-15 inhibited prophenoloxidase activation, but did not affect phenoloxidase activity, in B. mori hemolymph. Injection of recombinant Bmserpin-15 into B. mori larvae reduced significantly the transcript levels of antimicrobial peptides in fat body. Our results suggested that Bmserpin-15 plays an important role in the innate immunity of B. mori. PMID:25720980

  8. Cathepsin B protease is required for metamorphism in silkworm,Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    Gen-Hong Wang; Chun Liu; Qing-You Xia; Xing-Fu Zha; Jie Chen; Liang Jiang

    2008-01-01

    Cathepsin B belongs to lysosomal cysteine protease of the papain family.Temporal and spatial expression analysis of cathepsin B of Bombyx mori (BmCtB) was carried out based on Expression Sequence Tags (ESTs) data,oligonucleotide microarray,reverse transcription polymerase chain reaction (RT-PCR) and quantitative real-time PCR.Expression of BmCtB was observed in all of the tissues and stages.Among the 10 tested tissues,the fat body and posterior silk gland are the two most enriched tissues with BmCtB.During Bombyx development,there was an expression fastigium of BmCtB during metamorphosis.RNA interference was used to suppress the expression of cathepsin B during metamorphosis.Significant developmental defective phenotypes were obtained in the RNAi treated group.The dramatically reduced expression of BmCtB was confirmed by Northern blot and quantitative real-time PCR.These evidences strongly suggest cathepsin B proteinase was predominantly involved in the metabolism process of fat body and the posterior silk gland and was critical for metamorphism and development of silkworm,Bombyx mori.

  9. Genome-wide identification and characterization of ATP-binding cassette transporters in the silkworm, Bombyx mori

    OpenAIRE

    Zhang Jianzhen; Luan Yunxia; Guo Enen; Tian Ling; Zhou Shun; Liu Shumin; Li Sheng

    2011-01-01

    Abstract Background The ATP-binding cassette (ABC) transporter superfamily is the largest transporter gene family responsible for transporting specific molecules across lipid membranes in all living organisms. In insects, ABC transporters not only have important functions in molecule transport, but also play roles in insecticide resistance, metabolism and development. Results From the genome of the silkworm, Bombyx mori, we have identified 51 putative ABC genes which are classified into eight...

  10. Characterization of a ligand-gated cation channel based on an inositol receptor in the silkworm, Bombyx mori.

    Science.gov (United States)

    Kikuta, Shingo; Endo, Haruka; Tomita, Natsuo; Takada, Tomoyuki; Morita, Chiharu; Asaoka, Kiyoshi; Sato, Ryoichi

    2016-07-01

    Insect herbivores recognize non-volatile compounds in plants to direct their feeding behavior. Gustatory receptors (Gr) appear to be required for nutrient recognition by gustatory organs in the mouthparts of insects. Gr10 is expressed in Bombyx mori (BmGr10) mouthparts such as maxillary galea, maxillary palp, and labrum. BmGr10 is predicted to function in sugar recognition; however, the precise biochemical function remains obscure. Larvae of B. mori are monophagous feeders able to find and feed on mulberry leaves. Soluble mulberry leaf extract contains sucrose, glucose, fructose, and myo-inositol. In this study, we identified BmGr10 as an inositol receptor using electrophysiological analysis with the Xenopus oocyte expression system and Ca(2+) imaging techniques using mammalian cells. These results demonstrated that Xenopus oocytes or HEK293T cells expressing BmGr10 specifically respond to myo-inositol and epi-inositol but do not respond to any mono-, di-, or tri-saccharides or to some sugar alcohols. These inositols caused Ca(2+) and Na(+) influxes into the cytoplasm independently of a G protein-mediated signaling cascade, indicating that BmGr10 is a ligand-gated cation channel. Overall, BmGr10 plays an important role in the myo-inositol recognition required for B. mori larval feeding behavior. PMID:27132146

  11. Soaking RNAi in Bombyx mori BmN4-SID1 cells arrests cell cycle progression.

    Science.gov (United States)

    Mon, Hiroaki; Li, Zhiqing; Kobayashi, Isao; Tomita, Shuichiro; Lee, JaeMan; Sezutsu, Hideki; Tamura, Toshiki; Kusakabe, Takahiro

    2013-01-01

    RNA interference (RNAi) is an evolutionarily conserved mechanism for sequence-specific gene silencing. Previously, the BmN4-SID1 cell expressing Caenorhabditis ele gans SID-1 was established, in which soaking RNAi could induce effective gene silencing. To establish its utility, 6 cell cycle progression related cDNAs, CDK1, MYC, MYB, RNRS, CDT1, and GEMININ, were isolated from the silkworm, Bombyx mori L. (Lepidoptera: Bombycidae), and their expressions were further silenced by soaking RNAi in the BmN4-SID1 cells. The cell cycle progression analysis using flow cytometer demonstrated that the small amount of double stranded RNA was enough to arrest cell cycle progression at the specific cell phases. These data suggest that RNAi in the BmN4-SID1 cells can be used as a powerful tool for loss-of-function analysis of B. mori genes. PMID:24773378

  12. Structure of autophagy-related protein Atg8 from the silkworm Bombyx mori

    International Nuclear Information System (INIS)

    The crystal structure of autophagy-related protein Atg8 from the silkworm B. mori has two additional helices at the N-terminus before the expected ubiquitin fold. Autophagy-related protein Atg8 is ubiquitous in all eukaryotes. It is involved in the Atg8–PE ubiquitin-like conjugation system, which is essential for autophagosome formation. The structures of Atg8 from different species are very similar and share a ubiquitin-fold domain at the C-terminus. In the 2.40 Å crystal structure of Atg8 from the silkworm Bombyx mori reported here, the ubiquitin fold at the C-terminus is preceded by two additional helices at the N-terminus

  13. Expression of two types of acetylcholinesterase gene from the silkworm, Bombyx mori, in insect cells

    Institute of Scientific and Technical Information of China (English)

    JIN-YAN SHANG; YA-MING SHAO; GUO-JUN LANG; GAN YUAN; ZHEN-HUA TANG; CHUAN-XI ZHANG

    2007-01-01

    Complementary DNAs encoding two types of acetylcholinesterase(AChE)were isolated from the silkworm, Bombyx mori. The type 1 (Bmace1) and type 2 (Bmace2) ORFs are 2052 and 1917 bp in length, respectively. Both the complete ORFs of the Bmaces and Cterminal truncated forms were recombined into the Bacmid baculovirus vector under the control of the polyhedrin promoter and expressed in Trichoplusia ni (Tn-5B 1-4) cells. The resulting products exhibited AChE activity and glycosylation of the expressed proteins. An inhibition assay indicated that the ace2-type enzyme was more sensitive than the acel-type enzyme to inhibition by eserine and paraoxon.

  14. From genome to proteome: great progress in the domesticated silkworm (Bombyx mori L.)

    Institute of Scientific and Technical Information of China (English)

    Zhonghua Zhou; Huijuan Yang; Boxiong Zhong

    2008-01-01

    As the only truly domesticated insect,the silkworm not only has great economic value,but it also has value as a model for genetics and molecular biology research.Genomics and proteomics have recently shown vast potential to be essential tools in domesticated silkworm research,especially after the completion of the Bombyx mori genome sequence.This paper reviews the progress of the domesticated silkworm genome,particularly focusing on its genetic map,physical map and functional genome.This review also presents proteomics,the proteomic technique and its application in silkworm research.

  15. The biochemical effects of potassium chloride on the silkworm, ( Bombyx mori L.)

    Institute of Scientific and Technical Information of China (English)

    ARUNDHUTIBHATTACHARYA; BASSAPPAB.KALIWAL

    2005-01-01

    The supplementation with 50, 100 and 150μg/mL potassium chloride to the fifth instar larvae of the silkworm Bombyx mori on fat body glycogen, protein, total lipids and haemolymph protein and trehalose were analyzed. The fat body glycogen and protein and haemolymph protein were increased significantly in all the treated groups; whereas fat body total lipids increased only in 100 and 150μg/mL and haemolymph trehalose increased only in 150μg/mL potassium chloride-treated groups when compared with those of the corresponding parameters of the carrier controls.

  16. Correlation between yield and biochemical parameters in the mulberry silkworm,Bombyx mori L.

    Science.gov (United States)

    Chatterjee, S N; Rao, C G; Chatterjee, G K; Ashwath, S K; Patnaik, A K

    1993-11-01

    A detailed study was carried out on six biochemical parameters and four yield attributes using multiple regression analysis to investigate their relationship in the mulberry silkworm,Bombyx mori. The study generated new information on the importance of digestive amylase activity for the survival of the silkworm and revealed the inability of other enzymes to affect this relationship. Data also substantiate the observations made earlier on the genetic variability of amylase in the mulberry silkworm. Analyses extend the positive role of alkaline phosphatase and invertase in the expression of the other yield traits studied and indicate the definite possibility of using biochemical markers for silkworm breeding. PMID:24190267

  17. Expression, purification and characterization of methyl DNA binding protein from Bombyx mori

    OpenAIRE

    Uno, Tomohide; Nomura, Yuka; Nakamura, Masahiko; Nakao, Atsushi; Tajima, Shoji; Kanamaru, Kengo; Yamagata, Hiroshi; Iwanaga, Yousuke

    2005-01-01

    A cDNA clone encoding methyl DNA binding domain-containing protein (bMBD2/3) was obtained by homology searches using a Bombyx mori fat body cDNA library. The cDNA encoded a polypeptide with 249 amino acids sharing 54% similarity with the methyl DNA binding protein from Drosophila melanogaster. To characterize the biochemical properties of bMBD2/3, the clone was expressed in Escherichia coli as His-tagged protein. The recombinant protein was purified to homogeneity using Ni-NTA superflow resin...

  18. Hygroscopics and yield characteristics of cocoon in races of Bombyx mori L.

    OpenAIRE

    Antonio José Porto; José Eduardo de Almeida

    2012-01-01

    Cocoons, cocoon shells and chrysalis of four races of Bombyx mori L., two from China (C202 and C212) and two from Japan (B104 and M11-2), were analyzed in function of determinant factors as weight (dry mater and moisture) and by hygroscopic capacity after cooking. It was used a complete randomized design, with four treatments and ten replications. Although the cocoon shell has a higher hygroscopic capacity, the percentage of water in the chrysalis is the main factor to determine the weight of...

  19. Study on fibroin heavy chain of the silkworm Bombyx mori by fluorescence in situ hybridization (FISH)

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    The sericulture industry plays a very important role in our national economy. Silkworm (Bombyx mori) is always regarded as a model animal and biological reactor. There have been detailed studies on the structure, expression and control and molecular evolution of silk genes. However, few, if any, reports are available on the localization of structural genes in silkworm by molecular cytogenetics. The present experiment has tentatively localized the Fib-H gene at the distal end of the 25th linkage group, namely at the 25-0.0 position, and verified that Fib-H has only one locus, thus providing a temporary solution to the problem about its localization.

  20. Comparing the properties of Bombyx mori silk cocoons against sericin-fibroin regummed biocomposite sheets.

    Science.gov (United States)

    Morin, Alexander; Alam, Parvez

    2016-08-01

    This paper considers the utility of sericin, a degumming waste product, in the regumming of Bombyx mori silk fibroin fibres to form sericin-fibroin biocomposites. Regummed biocomposites have a chemical character that is somewhat closer to fibroin than sericin, though sericin presence is confirmed through FT-IR spectroscopy. Using direct measurements we further find the weight fractions of sericin in the regummed biocomposites and the native cocoons differ by only 5%. Mechanically, B. mori cocoons exhibit brittle stress-strain characteristics, failing at strengths of X̅= 16.6MPa and at strains of X̅= 13%. Contrarily, aligning fibroin fibres to a unidirectional axis in the regummed biocomposites causes them to exhibit characteristics of strain hardening, which is itself a typical characteristic of silk fibre pulled in tension. Though they are half as strong (X̅= 7.2MPa), regummed biocomposites are able to absorb five times more mechanical energy (X̅= 5.6MJm(-3)) than the B. mori cocoons (X̅= 1.1MJm(-3)) and are furthermore able to elongate to more than ten times (X̅= 180%) that of the native cocoons prior to failure. Our research shows that degummed B. mori cocoons can be regummed into sheets that have potential for use as load bearing engineering biocomposites. PMID:27157746

  1. Transmission Electron Microscopy of Bombyx Mori Silk Fibers

    Science.gov (United States)

    Shen, Y.; Martin, D. C.

    1997-03-01

    The microstructure of B. Mori silk fibers before and after degumming was examined by TEM, selected area electron diffraction (SAED), WAXS and low voltage SEM. SEM micrographs of the neat cocoon revealed a network of pairs of twisting filaments. After degumming, there were only individual filaments showing a surface texture consistent with an oriented fibrillar structure in the fiber interior. WAXS patterns confirmed the oriented beta-sheet crystal structure common to silkworm and spider silks. Low dose SAED results were fully consistent with the WAXS data, and revealed that the crystallographic texture did not vary significantly across the fiber diameter. TEM observations of microtomed fiber cross sections indicated a somewhat irregular shape, and also revealed a 0.5-2 micron sericin coating which was removed by the degumming process. TEM observations of the degummed silk fiber showed banded features with a characteristic spacing of nominally 600 nm along the fiber axis. These bands were oriented in a roughly parabolic or V-shape pointing along one axis within a given fiber. We hypothesize that this orientation is induced by the extrusion during the spinning process. Equatorial DF images revealed that axial and lateral sizes of the β-sheet crystallites in silk fibroin ranged from 20 to 170 nm and from 1 to 24 nm, respectively. Crazes developed in the degummed silk fiber parallel to the fiber direction. The formation of these crazes suggests that there are significant lateral interactions between fibrils in silk fibers.

  2. Preparation of Porous Scaffolds from Silk Fibroin Extracted from the Silk Gland of Bombyx mori (B. mori

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    Liangjun Zhu

    2012-06-01

    Full Text Available In order to use a simple and ecofriendly method to prepare porous silk scaffolds, aqueous silk fibroin solution (ASF was extracted from silk gland of 7-day-old fifth instar larvae of Bombyx mori (B. mori. SDS-page analysis indicated that the obtained fibroin had a molecular weight higher than 200 kDa. The fabrication of porous scaffolds from ASF was achieved by using the freeze-drying method. The pore of porous scaffolds is homogenous and tends to become smaller with an increase in the concentration of ASF. Conversely, the porosity is decreased. The porous scaffolds show impressive compressive strength which can be as high as 6.9 ± 0.4 MPa. Furthermore, ASF has high cell adhesion and growth activity. It also exhibits high ALP activity. This implies that porous scaffolds prepared from ASF have biocompatibility. Therefore, the porous scaffolds prepared in this study have potential application in tissue engineering due to the impressive compressive strength and biocompatibility.

  3. Construction of an in vivo system for functional analysis of the genes involved in sex pheromone production in the silkmoth, Bombyx mori.

    Directory of Open Access Journals (Sweden)

    Ken-IchiMoto

    2012-02-01

    Full Text Available Moths produce species-specific sex pheromones to attract conspecific mates. The biochemical processes that comprise sex pheromone biosynthesis are precisely regulated and a number of gene products are involved in this biosynthesis and regulation. In recent years, at least 300 EST clones have been isolated from Bombyx mori pheromone gland (PG specific cDNA libraries with some of those clones (i.e., B. mori PG-specific desaturase1 (Bmpgdesat1, PG-specific fatty-acyl reductase (pgFAR, PG-specific acyl-CoA-binding protein (pgACBP, B. mori fatty acid transport protein (BmFATP, B. mori lipid storage droplet protein-1 (BmLsd1 characterized and demonstrated to play a role in sex pheromone production. However, most of the EST clones have yet to be fully characterized and identified. To develop an efficient system for analyzing sex pheromone production-related genes, we investigated the feasibility of a novel gene analysis system using the upstream region of Bmpgdesat1 that should contain a PG-specific gene promoter in conjunction with piggyBac vector-mediated germ-line transformation. As a result, we have been able to obtain expression of our reporter gene (enhanced green fluorescent protein in the PG but not in other tissues of transgenic B. mori. Current results indicate that we have successfully constructed a novel in vivo gene analysis system for sex pheromone production in B. mori.

  4. Construction of an in vivo system for functional analysis of the genes involved in sex pheromone production in the silkmoth, Bombyx mori.

    Science.gov (United States)

    Moto, Ken-Ichi; Matsumoto, Shogo

    2012-01-01

    Moths produce species-specific sex pheromones to attract conspecific mates. The biochemical processes that comprise sex pheromone biosynthesis are precisely regulated and a number of gene products are involved in this biosynthesis and regulation. In recent years, at least 300 EST clones have been isolated from Bombyx mori pheromone gland (PG) specific cDNA libraries with some of those clones [i.e., B. mori PG-specific desaturase 1 (Bmpgdesat1), PG-specific fatty acyl reductase, PG-specific acyl-CoA-binding protein, B. mori fatty acid transport protein, B. mori lipid storage droplet protein-1] characterized and demonstrated to play a role in sex pheromone production. However, most of the EST clones have yet to be fully characterized and identified. To develop an efficient system for analyzing sex pheromone production-related genes, we investigated the feasibility of a novel gene analysis system using the upstream region of Bmpgdesat1 that should contain a PG-specific gene promoter in conjunction with piggyBac vector-mediated germ line transformation. As a result, we have been able to obtain expression of our reporter gene (enhanced green fluorescent protein) in the PG but not in other tissues of transgenic B. mori. Current results indicate that we have successfully constructed a novel in vivo gene analysis system for sex pheromone production in B. mori. PMID:22649415

  5. Heritable genome editing with CRISPR/Cas9 in the silkworm, Bombyx mori.

    Directory of Open Access Journals (Sweden)

    Wei Wei

    Full Text Available We report the establishment of an efficient and heritable gene mutagenesis method in the silkworm Bombyx mori using modified type II clustered regularly interspaced short palindromic repeats (CRISPR with an associated protein (Cas9 system. Using four loci Bm-ok, BmKMO, BmTH, and Bmtan as candidates, we proved that genome alterations at specific sites could be induced by direct microinjection of specific guide RNA and Cas9-mRNA into silkworm embryos. Mutation frequencies of 16.7-35.0% were observed in the injected generation, and DNA fragments deletions were also noted. Bm-ok mosaic mutants were used to test for mutant heritability due to the easily determined translucent epidermal phenotype of Bm-ok-disrupted cells. Two crossing strategies were used. In the first, injected Bm-ok moths were crossed with wild-type moths, and a 28.6% frequency of germline mutation transmission was observed. In the second strategy, two Bm-ok mosaic mutant moths were crossed with each other, and 93.6% of the offsprings appeared mutations in both alleles of Bm-ok gene (compound heterozygous. In summary, the CRISPR/Cas9 system can act as a highly specific and heritable gene-editing tool in Bombyx mori.

  6. Characterization of antiviral and antibacterial activity of Bombyx mori seroin proteins.

    Science.gov (United States)

    Singh, C P; Vaishna, R L; Kakkar, A; Arunkumar, K P; Nagaraju, J

    2014-09-01

    Lepidopterans as other insects have a very potent innate immune system, which basically comprises cellular and humoral defence mechanisms against bacterial and fungal infections. In lepidopterans, not much is known about the defence mechanisms against viral pathogens, such as baculoviruses. Here we show that small silk proteins of the domesticated silkworm, Bombyx mori, called seroins, act as antiviral agents against a baculovirus pathogen, Bombyx mori nucleopolyhedrovirus (BmNPV). Involvement of these proteins in the inhibition of baculovirus infection was revealed by estimating the viral load upon their dsRNA-mediated knockdown. Additionally, we found through antimicrobial assays that seroins are potent inhibitors of bacterial growth. Binding competition assays followed by antimicrobial assays showed that seroins bind to peptidoglycan, a cell wall component of bacteria. Analysis of bacterial load upon knockdown of seroins resulted in higher proliferation of bacteria. Phylogenetic analysis showed the recent origin of seroins in a few moth species and duplication only in Bombycids. The antiviral and antibacterial activity of seroins shown in this study using several biochemical and molecular biological assays provide strong evidence to characterize them as antimicrobial proteins. Hence, we hypothesize that seroins are potent candidates for use in development of transgene-based disease resistant silkworm strains. PMID:24628957

  7. Crystallization and preliminary X-ray analysis of cecropin B from Bombyx mori

    International Nuclear Information System (INIS)

    Cecropin B derived from the hemolymph of Bombyx mori has been crystallized by the hanging-drop vapour-diffusion method. The crystal diffracted to 1.43 Å resolution using X-ray radiation. Cecropin B is a 37-residue cationic antimicrobial peptide derived from the haemolymph of Bombyx mori. The precise mechanism by which cecropins exert their antimicrobial and cytolytic activities is not well understood. Crystals of cecropin B were obtained by the hanging-drop vapour-diffusion method using polyethylene glycol as a precipitant at 289 K. The crystal diffracted to 1.43 Å resolution using X-ray radiation and belonged to the orthorhombic space group P1, with unit-cell parameters a = 15.08, b = 22.75, c = 30.20 Å, α = 96.9, β = 103.1, γ = 96.5°. The asymmetric unit contained only one molecule of cecropin B, with a calculated Matthews coefficient of 2.48 Å3 Da−1 and a solvent content of 50.4%

  8. Intracellular transduction in the regulation of pheromone biosynthesis of the silkworm, Bombyx mori: suggested involvement of calmodulin and phosphoprotein phosphatase.

    Science.gov (United States)

    Matsumoto, S; Ozawa, R; Nagamine, T; Kim, G H; Uchiumi, K; Shono, T; Mitsui, T

    1995-03-01

    We have tested the effects of chemicals on bombykol production in vitro in the silkworm, Bombyx mori, to probe the biochemical steps as well as underlying mechanisms regulated by PBAN. These results suggest the involvement of calmodulin and phosphoprotein phosphatase in the intracellular signal transduction of PBAN action. PMID:7766202

  9. Construction of the Bac-to-Bac System of Bombyx mori Nucleopolyhedroviru

    Institute of Scientific and Technical Information of China (English)

    Jin-shan HUANG; Bi-fang HAO; Xiu-lian SUN; Fei DENG; Hua-lin WANG; Zhi-hong HU

    2007-01-01

    To construct the Bac-to-Bac expression system of Bombyx mori nucleopolyhedrovirus (BmNPV), a transfer vector was constructed which contained an Escherichia coli (E. coli) mini-F replicon and a lacZ: attTN7: lacZ cassette within the upstream and downstream regions of the BmNPV polyhedrin gene. B. mori larvae were cotransfected with wild-type BmNPV genomic DNA and the transfer vector through subcutaneous injection to generate recombinant viruses by homologous recombination in vivo. The genomic DNA of budded viruses extracted from the hemolymph of the transfected larvae was used to transform E. coli DH10B. Recombinant bacmids were screened by kanamycin resistance, PCR and restriction enzyme (REN) digestion. One of the bacmid colonies, BmBacJS13, which had similar REN profiles to that of wild-type BmNPV, was selected for further research. To investigate the infectivity of BmBacJS13, the polyhedrin gene was introduced into the bacmid and the resultant recombinant (BmBacJS13-ph) was transfected to BmN cells. The budded viruses were collected from the supernatant of the transfected cells and used for infecting BmN cells. Growth curve analysis indicated that BmBacJS13-ph had a similar growth curve to that of wild-type BmNPV. Bio-assays indicated that BmBacJS13-ph was also infectious to B. mori larvae.

  10. Metabolic allometry during development and metamorphosis of the silkworm Bombyx mori: analyses, patterns, and mechanisms.

    Science.gov (United States)

    Blossman-Myer, Bonnie L; Burggren, Warren W

    2010-01-01

    Intraspecific allometric (scaling) relationships for metabolism, which have received little examination compared to interspecific relationships, reflect a complex interplay of organogenesis, growth, and shifting physiologies. In this study of the silkworm Bombyx mori, we hypothesized that allometric relationships for metabolism both across all developmental stages and within each stage would not reflect conventional scaling coefficients (e.g., b not equal to 0.75). Histology, gross morphology, body surface and cross-sectional area, total lipid content, and cytochrome c oxidase activity levels (as evidence of the total metabolic potential of mitochondria) were determined across development. Also measured were oxygen consumption, carbon dioxide production, and the respiratory exchange ratio. The overall slope, b, in the allometric equation relating to body mass across all developmental stages was 0.82, not greatly different from the value of 0.75 typical of interspecific data. However, within larval instars II-V and in prepupae, b varied between 0.99 and 1.49, far higher than hypothesized. Thus, in B. mori, an analytical approach that lumps all developmental stages hides interinstar variability. Morphological and biochemical data suggest that observed scaling patterns in B. mori are likely correlated with changes in overall mitochondrial density rather than with specific changes in body proportion of tissues with higher intrinsic metabolic intensity. PMID:20105069

  11. Cloning, expression and enzymatic properties analysis of dihydrofolate reductase gene from the silkworm, Bombyx mori.

    Science.gov (United States)

    Wang, Wenjing; Gao, Junshan; Wang, Jing; Liu, Chaoliang; Meng, Yan

    2012-12-01

    Tetrahydrobiopterin (BH4) is an essential cofactor for aromatic acid hydroxylases, which control the levels of monoamine neurotransmitters. BH4 deficiency has been associated with many neuropsychological disorders. Dihydrofolate reductase (DHFR) can catalyze 7,8-dihydrobiopterin to 5,6,7,8-tetrahydrobiopterin (BH4) in the salvage pathway of BH4 synthesis from sepiapterin (SP), a major pigment component contained in the integument of silkworm Bombyx mori mutant lemon (lem) in high concentration. In this study, we report the cloning of DHFR gene from the silkworm B. mori (BmDhfr) and identification of enzymatic properties of BmDHFR. BmDhfr is located on scaffold Bm_199 with a predicted gene model BGIBMGA013340, which encodes a 185-aa polypeptide with a predicted molecular mass of about 21 kDa. Biochemical analyses showed that the recombinant BmDHFR protein exhibited high enzymatic activity and suitable parameters to substrate. Together with our previous studies on SP reductase of B. mori (BmSPR) and the lem mutant, it may be an effective way to industrially extract SP from the lem silkworms in large scale to produce BH4 in vitro by co-expressing BmSPR and BmDHFR and using the extracted SP as a substrate in the future. PMID:23065260

  12. Expression of the Japanese oak silkworm Antheraea yamamai fibroin gene in the domesticated silkworm Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    Isao Kobayashi; Katsura Kojima; Hideki Sezutsu; Keiro Uchino; Toshiki Tamura

    2009-01-01

    To understand the evolutionary conservation of the gene expression mechanism and secretion machinery between Antheraea and Bombyx fibroins, we introduced the genomic A. yamamai fibroin gene into the domesticated silkworm, B. mori. The spliced A. yamamai fibroin mRNA appeared only in the posterior region of the silk gland of the transgenic silkworm, suggesting that the functions of the fibroin promoter region and the splicing machinery are conserved between these two species. The A. yamamai fibroin protein was detected in the lumen of the silk gland of the transgenic silkworm, albeit at lower levels compared with the B. mori-type fibroin. We found a strong degeneration of the posterior region of the silk gland of the transgenic silkworm. As a result, the cocoon shell weight was much lower in the transgenic silkworm than in the non-transgenic line. These results indicate that the promoter function and splicing machinery are well conserved between A. yamamai and B. mori but that the secretion mechanism of fibroin is diversified between the two.

  13. Relationship between Biological Characteristics of Beauveria bassiana (Bals.) Vuill and Pathogenicity to Bombyx mori L.%Relationship between Biological Characteristics of Beauveria bassiana (Bals.) Vuill and Pathogenicity to Bombyx mori L.

    Institute of Scientific and Technical Information of China (English)

    Haiyu LUO; Yecheng DENG; Yongmei LIAO; Ruiyu LI

    2012-01-01

    [Objective] This study was to investigate the relationship between biological characteristics of Beauveria bassiana (Bals.) Vuill and pathogenicity to Bombyx rnori L, with the aim to provide scientific basis for the control of white muscardine in Bombyx mori L. [Method] The strains isolated and purified from the 6 Beauveria bassiana biocontrol agents from all over the country and the 3 white muscardine silkworms collected from Guangxi provincial silkworm rearing areas were identified by the morphological observation and molecular biology technology. The pathogenicity of B. bassaina to silkworms was determined, and the biological characteristics such as growth diameter, sporulation and the extracellular protease activity of the different B. bassiana strains were compared. [Result] The isolated 9 strains were all B. bassaina (Bals.) Vuillemin, and all strains had high pathogenicity to silkworm, but with different pathogenicities. The growth diameter, sporulation and extracellular protease activity of different B. bassiana strains were also different, and showed correlation with the patheogenicity to silkworms. [Conclusion] B. bassiana spores production amount and exocellular protease activity had significant positive correlation with their pathogenicity to silkworm.

  14. Novel Infection System of Recombinant BmBDV DNA into BmN Cells of Silkworm, Bombyx mori.

    Science.gov (United States)

    Guo, Rui; Cao, Guangli; Zhu, Yuexiong; Kumar, Dhiraj; Xue, Renyu; Lu, Yahong; Hu, Xiaolong; Gong, Chengliang

    2016-10-01

    Bombyx mori bidensovirus (BmBDV) was previously termed as Bombyx mori densovirus type 2 and later it was reclassified in the new genus bidensovirus of the new family Bidnaviridae. The genome of BmBDV Zhenjiang isolate (BmBDV-Z) consists of two non-homologous single-stranded linear DNA molecules VD1 and VD2 which are encapsidated into separate virion. To investigate the infectivity of BmBDV DNA, recombinant plasmids pGEM-VD1 inserted with VD1 genome were transfected into the BmN cells of silkworm. Structural proteins of BmBDV were detected with Western blot and immunofluorescence assay, which indicates pGEM-VD1 replicated in the transfected BmN cells and viral proteins were also expressed. Through TEM observation, we identified about 20 nm BmBDV-like viral particles, which confirmed that BmBDV can be generated after transfection. Subsequently, a recombinant baculovirus BmBac-VD1 inserted with VD1 genome was constructed. Results of Western blot and immunofluorescence assay indicated that viral structural proteins of BmBDV were expressed in the BmBac-VD1-infected cells. Baculiform and spherical virions were also observed in infected cells by TEM, and two kinds of virions were separated. However, results of molecular biological detection revealed that infectious sequence from BmBac-VD1 was packaged within spherical virion. Therefore, we suggested that vector inserted with BmBDV genomic DNA showed infectivity, and BmBDV-like viral particles packaging recombinant DNA can be produced in the cultured BmN cells. Outcome of our current research provided not only a new method of infection to explore the gene function of BmBDV in vitro but also a protocol to facilitate development of more effective new-type pesticides. PMID:27447797

  15. Comparison of susceptibility of Chilo suppressalis and Bombyx mori to five Bacillus thuringiensis proteins.

    Science.gov (United States)

    Jiao, Yaoyu; Yang, Yan; Meissle, Michael; Peng, Yufa; Li, Yunhe

    2016-05-01

    Transformation of rice with genes encoding insecticidal Cry proteins from Bacillus thuringiensis (Bt) should confer high resistance to target lepidopteran pests, such as Chilo suppressalis, and low toxicity to non-target organisms, such as silkworm Bombyx mori. Five purified Cry proteins that have been used for plant transformation were tested using dietary exposure assays. The susceptibility of C. suppressalis larvae to the five insecticidal proteins in the decreasing order was: Cry1Ca>Cry1Ab>Cry1Ac>Cry2Aa>Cry1Fa. However, the toxicities of the Cry proteins to B. mori were in the order: Cry1Fa>Cry1Ca>Cry2Aa>Cry1Ab>Cry1Ac. The Cry1Ca, Cry1Ab and Cry1Ac proteins exhibited relatively high toxicity to C. suppressalis larvae, with EC50 values of 16.4, 45.8 and 89.6ng/g, respectively. The toxicities of the three Cry proteins to B. mori larvae were 8, 14, and 22times lower, with EC50 values of 138.3, 628.4 and 1939.2ng/g, respectively. The Cry1Fa and Cry2Aa proteins showed high toxicity to B. mori larvae, with EC50 values of 135.7 and 373.9ng/g, respectively, but low toxicity to C. suppressalis larvae, with EC50 values of 6092.1 and 1208.5ng/g, respectively. We thus conclude that Cry1Ab, Cry1Ac and Cry1Ca are appropriate for transforming rice to control lepidopteran rice pests. In contrast, Cry1Fa and Cry2Aa are not appropriate due to their high toxicity to silkworm larvae and low activity against the target pest. PMID:26994840

  16. Characterization of Klebsiella granulomatis pathogenic to silkworm, Bombyx mori L.

    OpenAIRE

    Mohanta, M. K.; Saha, A. K.; Saleh, D. K. M. A.; M S Islam; Mannan, K. S. B.; Fakruddin, M.

    2014-01-01

    Bacterial disease of silkworm causes significant reduction of silk production leading to huge economic loss. This study aims to isolate bacteria from diseased silkworm and to determine its pathogenicity and antibiotic resistance. A strain of Klebsiella granulomatis has been isolated from silkworm haemolymph which was later identified on the basis of biochemical tests and 16S rRNA gene sequencing. The optimum culture condition of K. granulomatis was determined at pH 7.0 and 37 °C temperature. ...

  17. Genome-wide identification and expression analysis of serine proteases and homologs in the silkworm Bombyx mori

    Directory of Open Access Journals (Sweden)

    Xu Ping-Zhen

    2010-06-01

    Full Text Available Abstract Background Serine proteases (SPs and serine proteases homologs (SPHs are a large group of proteolytic enzymes, with important roles in a variety of physiological processes, such as cell signalling, defense and development. Genome-wide identification and expression analysis of serine proteases and their homologs in the silkworm might provide valuable information about their biological functions. Results In this study, 51 SP genes and 92 SPH genes were systematically identified in the genome of the silkworm Bombyx mori. Phylogenetic analysis indicated that six gene families have been amplified species-specifically in the silkworm, and the members of them showed chromosomal distribution of tandem repeats. Microarray analysis suggests that many silkworm-specific genes, such as members of SP_fam12, 13, 14 and 15, show expression patterns that are specific to tissues or developmental stages. The roles of SPs and SPHs in resisting pathogens were investigated in silkworms when they were infected by Escherichia coli, Bacillus bombysepticus, Batrytis bassiana and B. mori nucleopolyhedrovirus, respectively. Microarray experiment and real-time quantitative RT-PCR showed that 18 SP or SPH genes were significantly up-regulated after pathogen induction, suggesting that SP and SPH genes might participate in pathogenic microorganism resistance in B. mori. Conclusion Silkworm SP and SPH genes were identified. Comparative genomics showed that SP and SPH genes belong to a large family, whose members are generated mainly by tandem repeat evolution. We found that silkworm has species-specific SP and SPH genes. Phylogenetic and microarray analyses provide an overview of the silkworm SP and SPHs, and facilitate future functional studies on these enzymes.

  18. Structure of Cytoplasmic Polyhedrosis Virus from Bombyx mori.

    Science.gov (United States)

    Yu, Xue-Kui; Lu, Xin-Yin; Zhang, Hong; Zhou, Zhen-Hong; Zhang, Qin-Fen; Zhang, Xing; Zhang, Jing-Qiang

    1999-01-01

    The structures of full and empty capsids of CPV were studied by negative staining and electron cryomicroscopy and computer reconstruction techniques. By comparing the structures and biochemical compositions, the CPV was identified as a single-layered capsid with its five structural proteins located on it. This single capsid is arranged according to T=1 icosahedral symmetry with 12 turret-like spikes at its icosahedral vertices. The empty and full CPV show identical capsid but differ inside. The dense and ordered genomic dsRNA is located inside the full CPV. The internal space of the empty CPV has almost no electron density except for 12 electron densities attributed the transcriptional enzyme complexes extending inward from the base part of CPV spikes. PMID:12114971

  19. Silkworm Gut Fiber of Bombyx mori as an Implantable and Biocompatible Light-Diffusing Fiber

    Directory of Open Access Journals (Sweden)

    Jose Luis Cenis

    2016-07-01

    Full Text Available This work describes a new approach to the delivery of light in deeper tissues, through a silk filament that is implantable, biocompatible, and biodegradable. In the present work, silkworm gut fibers (SGFs of Bombyx mori L., are made by stretching the silk glands. Morphological, structural, and optical properties of the fibers have been characterized and the stimulatory effect of red laser light diffused from the fiber was assayed in fibroblast cultures. SGFs are formed by silk fibroin (SF mainly in a β-sheet conformation, a stable and non-soluble state in water or biological fluids. The fibers showed a high degree of transparency to visible and infrared radiation. Using a red laser (λ = 650 nm as source, the light was efficiently diffused along the fiber wall, promoting a significant increment in the cell metabolism 5 h after the irradiation. SGFs have shown their excellent properties as light-diffusing optical fibers with a stimulatory effect on cells.

  20. Silkworm Gut Fiber of Bombyx mori as an Implantable and Biocompatible Light-Diffusing Fiber

    Science.gov (United States)

    Cenis, Jose Luis; Aznar-Cervantes, Salvador D.; Lozano-Pérez, Antonio Abel; Rojo, Marta; Muñoz, Juan; Meseguer-Olmo, Luis; Arenas, Aurelio

    2016-01-01

    This work describes a new approach to the delivery of light in deeper tissues, through a silk filament that is implantable, biocompatible, and biodegradable. In the present work, silkworm gut fibers (SGFs) of Bombyx mori L., are made by stretching the silk glands. Morphological, structural, and optical properties of the fibers have been characterized and the stimulatory effect of red laser light diffused from the fiber was assayed in fibroblast cultures. SGFs are formed by silk fibroin (SF) mainly in a β-sheet conformation, a stable and non-soluble state in water or biological fluids. The fibers showed a high degree of transparency to visible and infrared radiation. Using a red laser (λ = 650 nm) as source, the light was efficiently diffused along the fiber wall, promoting a significant increment in the cell metabolism 5 h after the irradiation. SGFs have shown their excellent properties as light-diffusing optical fibers with a stimulatory effect on cells. PMID:27438824

  1. Transcription factor SGF1 is critical for the neurodevelopment in the silkworm, Bombyx mori.

    Science.gov (United States)

    Liu, Zhao-Yang; Yu, Qi; Yang, Chun-Hong; Meng, Miao; Ren, Chun-Jiu; Mu, Zhi-Mei; Cui, Wei-Zheng; Liu, Qing-Xin

    2016-08-01

    FoxA transcription factors play vital roles in regulating the expression of organ-specific genes. BmSGF1, the sole FoxA family member in Bombyx mori, is required for development of the silk gland. However, the function of BmSGF1 in development of the nervous system in the silkworm remains unknown. Here, we show that the amino acids sequence of BmSGF1 is evolutionarily conserved in its middle region from Trichoplax adhaerens to human and diverged from the homologues in most other species in its N-terminal region. BmSGF1 expresses in the nervous system at the embryonic stage. Knockdown of Bmsgf1 by RNA interference (RNAi) results in abnormal development of axons. Therefore, our results demonstrate that BmSGF1 is an indispensable regulator for neurodevelopment. PMID:27106119

  2. The complete mitochondrial genome of Bombyx mori strain Yu39 (Lepidoptera: Bombycidae).

    Science.gov (United States)

    Zhang, Yong-Liang; Zhao, Jin-Hui; Zhou, Qi-Ming

    2016-09-01

    The complete mitochondrial genome of Bombyx mori strain Yu39 (Lepidoptera: Bombycidae) is a circular molecule of 15,652 bp in length, containing 37 typical mitochondrial genes: 13 protein-coding genes (PCGs), 22 transfer RNAs, 2 ribosomal RNAs and a non-coding AT-rich region. Its gene order and arrangement are identical to the common type found in most insect mitogenomes. All PCGs start with a typical ATN codon, except for the cox1 gene, which begins with uncertained codon. All PCGs terminate in the common stop codon TAA, except for the cox1 and cox2, which use single T as their stop codons. The non-coding AT-rich region is 494-bp long, located between rrnS and trnM genes. It contains some structures of repeated motifs and microsatellite-like elements characteristic of the other lepidopterons. PMID:25676361

  3. Analysis of defective genomes of bombyx mori nucleopolyhedrovirus generated by serial undiluted passage in cell culture

    International Nuclear Information System (INIS)

    Viral DNA was extracted from cells infected with bombyx mori nucleopolyhedrovirus (BmNPV) Dl strain after 34 serial undiluted passages (P34). P34 DNA was subjected to restriction analysis and Southern blot hybridisation using standard D 1 DNA and P34 DNA of BmNPV as probes. Based on hybridisation profiles, the BmNPV DNA regions retained in the P34 DNA were localised on HindIII and PstI restriction maps. Two regions of BmNPV DNA located at 0 - 12.8 and 40.2 - 65.0 map unit were highly conserved in P34 DNA. These regions contained two of three interspersed homologous sequences, but only one of five homologous regions. This suggests that ihss may have an essential role in BmNPV replication. (authors)

  4. Biochemical properties of an omega-class glutathione S-transferase of the silkmoth, Bombyx mori.

    Science.gov (United States)

    Yamamoto, Kohji; Nagaoka, Sumiharu; Banno, Yutaka; Aso, Yoichi

    2009-05-01

    A cDNA encoding an omega-class glutathione S-transferase of the silkmoth, Bombyx mori (bmGSTO), was cloned by reverse transcriptase-polymerase chain reaction. The resulting clone was sequenced and deduced for amino acid sequence, which revealed 40, 40, and 39% identities to omega-class GSTs from human, pig, and mouse, respectively. A recombinant protein (rbmGSTO) was functionally overexpressed in Escherichia coli cells in a soluble form and purified to homogeneity. rbmGSTO was able to catalyze the biotranslation of glutathione with 1-chloro-2,4-dinitrobenzene, a model substrate for GST, as well as with 4-hydroxynonenal, a product of lipid peroxidation. This enzyme was shown to have high affinity for organophosphorus insecticide and was present abundantly in silkmoth strain exhibiting fenitrothion resistance. These results indicate that bmGSTO could be involved in the increase in level of insecticide resistance for lepidopteran insects. PMID:19022397

  5. Identification and characterization of globin genes from two lepidopteran insects, Bombyx mori and Samia cynthia ricini.

    Science.gov (United States)

    Kawaoka, Shinpei; Katsuma, Susumu; Meng, Yan; Hayashi, Nobumitsu; Mita, Kazuei; Shimada, Toru

    2009-02-15

    We describe the characterization of hemoglobin-like genes from two lepidopteran insects, Bombyx mori (Bmglobin) and Samia cynthia ricini (Scglobin). Bmglobin and Scglobin are predicted to be intracellular proteins and contain amino acids required for heme and oxygen binding. Expression profiles of two lepidopteran globins, especially Bmglobin, were different from that of other insect globins. Although other insect globins are mainly associated with the tracheal system, Bmglobin was expressed almost exclusively in the Malpighian tubules, and the strongest signal for Scglobin was detected in the fat body. Furthermore, biochemical fractionation analysis revealed that both Bmglobin and Scglobin were localized in the cytoplasm. These results suggest that each lepidopteran globin has a distinct role in the tissues in which it is expressed and that the functions of insect globins are more divergent than previously thought. PMID:19059317

  6. Molecular and biochemical characterization of manganese-containing superoxide dismutase from the silkworm, Bombyx mori.

    Science.gov (United States)

    Yamamoto, Kohji; Zhang, Pingbo; He, Ningjia; Wang, Yongqiang; Aso, Yoichi; Banno, Yutaka; Fujii, Hiroshi

    2005-12-01

    Superoxide dismutase (SOD) is responsible for the removal of superoxide anion from living organisms. In this study, cDNA encoding the manganese-containing SOD (MnSOD) from the silkworm, Bombyx mori, was isolated by reverse transcriptase-polymerase chain reaction and sequenced. The deduced amino acid sequence of the MnSOD revealed 62% identity to that of the Drosophila melanogaster; both were close to each other in a phylogenetic tree. The MnSOD was overproduced in Escherichia coli and purified. The internal structure of the recombinant MnSOD was confirmed by peptide mass fingerprinting method. The recombinant MnSOD facilitating the reduction reaction of superoxide anion retained 75% of its original activity after incubation at pH 4-11 for 24 h at 4 degrees C. Its activity was never affected by incubation at pH 7 for 30 min below 50 degrees C. PMID:16236537

  7. Analysis of the structure and expression of the 30K protein genes in silkworm, Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    QUAN SUN; PING ZHAO; YING LIN; YONG HOU; QING-YOU XIA; ZHONG-HUAI XIANG

    2007-01-01

    A group of lipoproteins with molecular sizes of approximately 30 kDa, referredto as 30K proteins, are synthesized in fat body cells in the fifth instar larvae of silkworm,Bombyx mori. Analyzing the silkworm genome and its expressed sequence tags (ESTs), wefound 10 genes encoding 30K proteins, which are mainly distributed in three subfamilies.Of these, seven coding proteins were found to harbor the degrading sites of 30kP proteaseA, although the number of degrading sites may be different. As some potential corepromoters and regulatory elements were supposed to be essential for gene transcription, theexpression profiles of these genes were examined by semi-quantitative reverse transcriptionpolymerase chain reaction. Eight 30K protein genes were detected to express luxuriantly inthe fat body, while two were hardly expressed. Such results suggest that these 30K proteinsmay have different functions, and their adjacent regulatory elements play a crucial role inregulating their transcription.

  8. Hygroscopics and yield characteristics of cocoon in races of Bombyx mori L.

    Directory of Open Access Journals (Sweden)

    Antonio José Porto

    2012-01-01

    Full Text Available Cocoons, cocoon shells and chrysalis of four races of Bombyx mori L., two from China (C202 and C212 and two from Japan (B104 and M11-2, were analyzed in function of determinant factors as weight (dry mater and moisture and by hygroscopic capacity after cooking. It was used a complete randomized design, with four treatments and ten replications. Although the cocoon shell has a higher hygroscopic capacity, the percentage of water in the chrysalis is the main factor to determine the weight of cocoon. The percentage of water in the cocoon and its components are inversely related to the absorption after cooking. Of all races evaluated, the chinese ones had lower percentage of water in the cocoon and higher cocoon shell weight than the japanese races.

  9. Bacillus thuringiensis Cry1Aa toxin-binding region of Bombyx mori aminopeptidase N.

    Science.gov (United States)

    Yaoi, K; Nakanishi, K; Kadotani, T; Imamura, M; Koizumi, N; Iwahana, H; Sato, R

    1999-12-17

    The Bacillus thuringiensis Cry1Aa toxin-binding region of Bombyx mori aminopeptidase N (APN) was analyzed, to better understand the molecular mechanism of susceptibility to the toxin and the development of resistance in insects. APN was digested with lysylendopeptidase and the ability of the resulting fragments to bind to Cry1Aa and 1Ac toxins was examined. The binding abilities of the two toxins to these fragments were different. The Cry1Aa toxin bound to the fragment containing 40-Asp to 313-Lys, suggesting that the Cry1Aa toxin-binding site is located in the region between 40-Asp and 313-Lys, while Cry1Ac toxin bound exclusively to mature APN. Next, recombinant APN of various lengths was expressed in Escherichia coli cells and its ability to bind to Cry1Aa toxin was examined. The results localized the Cry1Aa toxin binding to the region between 135-Ile and 198-Pro. PMID:10606725

  10. Attack behavior of Podisus rostralis (Heteroptera: Pentatomidade adults on caterpillars of Bombyx mori (Lepidoptera: Bombycidae

    Directory of Open Access Journals (Sweden)

    Walkymário Paulo Lemos

    2005-11-01

    Full Text Available Attack behavior of the predator Podisus rostralis (Stäl (Heteroptera: Pentatomidae adults on fourth instar Bombyx mori L. (Lepidoptera: Bombycidae caterpillars was studied in laboratory conditions. Ten 24 hours old adults of this predator were observed during two hours with the following attack behavior: (1 Predator: prey finding; prey observation; touching prey with antenna; attack behavior; prey paralysis; predator retreat after attack; attack cessation; successive attacks; and (2 Prey: defense. The predator P. rostralis found its prey before attacking and it approached it with slow circular movements. The attack was usually made in the posterior part of the prey to reduce defense reaction. Larger size of prey in relation to the predator resulted difficult prey paralysis but it occurred in less than two hours.Estudou-se, em laboratório, o comportamento de ataque de adultos do predador Podisus rostralis (Stäl (Heteroptera: Pentatomidae tendo como presa lagartas de quarto estádio de Bombyx mori L. (Lepidoptera: Bombycidae. Dez adultos do predador, com 24 horas de idade, foram observados durante duas horas acompanhando-se os seguintes comportamentos de ataque: (1 Predador: localização da presa; observação da presa; toque das presas com as antenas; comportamento de ataque; paralisação da presa; fuga do predador após ataque; finalização do ataque; ataques sucessivos; e (2 Presa: defesa. O predador P. rostralis localizou sua presa antes do ataque, aproximando-se dela através de lentos movimentos circulares. O ataque é, usualmente, realizado na parte posterior da presa para reduzir reação de defesa. O maior tamanho da presa em relação ao predador pode dificultar a paralisação, porém o predador consegue paralisá-la em menos de duas horas.

  11. Characterisation of a desmosterol reductase involved in phytosterol dealkylation in the silkworm, Bombyx mori.

    Directory of Open Access Journals (Sweden)

    Leonora F Ciufo

    Full Text Available Most species of invertebrate animals cannot synthesise sterols de novo and many that feed on plants dealkylate phytosterols (mostly C(29 and C(28 yielding cholesterol (C(27. The final step of this dealkylation pathway involves desmosterol reductase (DHCR24-catalysed reduction of desmosterol to cholesterol. We now report the molecular characterisation in the silkworm, Bombyx mori, of such a desmosterol reductase involved in production of cholesterol from phytosterol, rather than in de novo synthesis of cholesterol. Phylogenomic analysis of putative desmosterol reductases revealed the occurrence of various clades that allowed for the identification of a strong reductase candidate gene in Bombyx mori (BGIBMGA 005735. Following PCR-based cloning of the cDNA (1.6 kb and its heterologous expression in Saccharomyces cerevisae, the recombinant protein catalysed reduction of desmosterol to cholesterol in an NADH- and FAD-dependent reaction.Conceptual translation of the cDNA, that encodes a 58.9 kDa protein, and database searching, revealed that the enzyme belongs to an FAD-dependent oxidoreductase family. Western blotting revealed reductase protein expression exclusively in the microsomal subcellular fraction and primarily in the gut. The protein is peripherally associated with microsomal membranes. 2D-native gel and PAGE analysis revealed that the reductase is part of a large complex with molecular weight approximately 250 kDa. The protein occurs in midgut microsomes at a fairly constant level throughout development in the last two instars, but is drastically reduced during the wandering stage in preparation for metamorphosis. Putative Broad Complex transcription factor-binding sites detectable upstream of the DHCR24 gene may play a role in this down-regulation.

  12. Novel non-autonomous transposable elements onWchromosome of the silkworm, Bombyx mori

    Indian Academy of Sciences (India)

    Hiroaki Abe; Tsuguru Fujii; Toru Shimada; Kazuei Mita

    2010-09-01

    The sex chromosomes of the silkworm Bombyx mori are designated ZW(XY) for females and ZZ (XX) for males. Numerous long terminal repeat (LTR) and non-LTR retrotransposons, retroposons and DNA transposons have accumulated as strata on the W chromosome. However, there are nucleotide sequences that do not show the characteristics of typical transposable elements on the W chromosome. To analyse these uncharacterized nucleotide sequences on the W chromosome, we used whole-genome shotgun (WGS) data and assembled data that was obtained using male genome DNA. Through these analyses, we found that almost all of these uncharacterized sequences were non-autonomous transposable elements that do not fit into the conventional classification. It is notable that some of these transposable elements contained the Bombyx short interspersed element (Bm1) sequences in the elements. We designated them as secondary-Bm1 transposable elements (SBTEs). Because putative ancestral SBTE nucleotide sequences without Bm1 do not occur in theWGS data, we suggest that the Bm1 sequences of SBTEs are not carried on each element merely as a package but are components of each element. Therefore, we confirmed that SBTEs should be classified as a new group of transposable elements.

  13. Characterization and expression patterns of let-7 microRNA in the silkworm (Bombyx mori

    Directory of Open Access Journals (Sweden)

    Hong Kaili

    2007-07-01

    Full Text Available Abstract Background lin-4 and let-7, the two founding members of heterochronic microRNA genes, are firstly confirmed in Caenorhabditis elegans to control the proper timing of developmental programs in a heterochronic pathway. let-7 has been thought to trigger the onset of adulthood across animal phyla. Ecdysone and Broad-Complex are required for the temporal expression of let-7 in Drosophila melanogaster. For a better understanding of the conservation and functions of let-7, we seek to explore how it is expressed in the silkworm (Bombyx mori. Results One member of let-7 family has been identified in silkworm computationally and experimentally. All known members of this family share the same nucleotides at ten positions within the mature sequences. Sequence logo and phylogenetic tree show that they are not only conserved but diversify to some extent among some species. The bmo-let-7 was very lowly expressed in ova harvested from newborn unmated female adult and in individuals from the first molt to the early third instar, highly expressed after the third molt, and the most abundant expression was observed after mounting, particularly after pupation. The expression levels were higher at the end of each instar and at the beginning of each molt than at other periods, coinciding with the pulse of ecdysone and BR-C as a whole. Using cultured ovary cell line, BmN-SWU1, we examined the effect of altered ecdysone levels on bmo-let-7 expression. The expression was also detected in various tissues of day 3 of the fifth instar and of from day 7 of the fifth to pupa, suggesting a wide distributing pattern with various signal intensities. Conclusion bmo-let-7 is stage- and tissue-specifically expressed in the silkworm. Although no signals were detected during embryonic development and first larval instar stages, the expression of bmo-let-7 was observed from the first molt, suggesting that it might also function at early larval stage of the silkworm. The

  14. cDNA cloning and expression of Bacillus thuringiensis Cry1Aa toxin binding 120 kDa aminopeptidase N from Bombyx mori.

    Science.gov (United States)

    Yaoi, K; Nakanishi, K; Kadotani, T; Imamura, M; Koizumi, N; Iwahana, H; Sato, R

    1999-01-18

    Bacillus thuringiensis Cry1Aa toxin binds to a 120 kDa putative receptor protein in the Bombyx mori midgut. Recently, this protein was purified and identified as glycosyl-phosphatidylinositol (GPI) anchored aminopeptidase N (APN). In this study, a full-length cDNA thought to encode this 120 kDa APN was isolated and sequenced. It has a 2958 bp ORF encoding 986 amino acids. In the deduced amino acid sequence, we identified GPI-anchor and zinc-metallopeptidase signals, which are the same as those of APNs of other insects that are reported to be putative Cry1 toxin receptors. The B. mori APN amino acid sequence also has a high similarity with those of the other APNs. Subsequently, the recombinant APN was expressed by Escherichia coli and its Cry1Aa toxin binding ability was analyzed. Ligand blotting showed that Cry1Aa toxin bound to the recombinant APN. PMID:9931470

  15. Control of protein synthesis in the female pupa of Bombyx mori

    International Nuclear Information System (INIS)

    For the purpose of understanding the mechanisms of insect metamorphosis, protein synthesis by silkmoth pupae has been studied. Synthetic rate and contents of total RNA and protein changed markedly in the female pupae of Bombyx mori. Attempt was made to find what the limiting step for the synthesis of the bulk of proteins during the adult development of female pupae is. Several female pupae of hydridstrain were homogenized at each of stated periods in buffer. The ribosomal fraction prepared from the homogenates was incubated in the buffer containing 3H-leucine or 3H-phenylalanine. The incorporation of leucine depending on endogenous mRNA and that of phenylalanine directed by added poly U were the largest in 9--10 days and 7th day, respectively. From the results, the synthesis of protein during the late adult development of female silkworms is controlled at the level of mRNA. The increase of ribosomes, which were active to bind mRNA, preceded the appearance of available endogenous mRNA, and it may be attributed to neogenesis and ''run-off'' of previous ribosomes. It is conceivable that such neogenesis or run-off serves as less direct control for the protein synthesis during the metamorphosis of Bombix mori. (Kobatake, H.)

  16. Molecular analysis of sex chromosome-linked mutants in the silkworm Bombyx mori

    Indian Academy of Sciences (India)

    Tsuguru Fujii; Hiroaki Abe; Toru Shimada

    2010-09-01

    In Bombyx mori, the W chromosome determines the female sex. A few W chromosome-linked mutations that cause masculinization of the female genitalia have been found. In female antennae of a recently isolated mutant, both female-type and male-type Bmdsx mRNAs were expressed, and BmOr1 (bombykol receptor) and BmOr3 (bombykal receptor), which are predominantly expressed in the antennae of male moths, were expressed about 50 times more abundantly in the antennae of mutant females than in those of normal females. These mutants are valuable resources for the molecular analysis of the sex-determination system. Besides the Fem gene, the quantitative egg size-determining gene Esd is thought to be present on the W chromosome, based on the observation that ZWW triploid moths produce larger eggs than ZZW triploids. The most recently updated B. mori genome assembly comprises 20.5 Mb of Z chromosome sequence. Using these sequence data, responsible genes or candidate genes for four Z-linked mutants have been reported. The od (distinct oily) and spli (soft and pliable) are caused by mutation in BmBLOS2 and Bmacj6, respectively. Bmap is a candidate gene for $V_g$ (vestigial). Similarly, Bmprm is a candidate gene for Md (muscle dystrophy), causing abnormal development of indirect flight muscle.

  17. Primary and secondary structure of 5.8S rRNA from the silkgland of Bombyx mori.

    OpenAIRE

    Fujiwara, H.; Kawata, Y.; H. Ishikawa

    1982-01-01

    Nucleotide sequence of 5.8S rRNA of the silkworm, Bombyx mori has been determined by gel sequencing methods. The 5.8S rRNA was the longest so far reported, with the 5'-terminal sequence several nucleotides longer than those of the other organisms. Upon constructing the secondary structure in accordance with the "burp gun" model (12), the Bombyx 5.8S rRNA formed a wide-open "muzzle" due to several unpaired bases at the ends. The overall structure also appeared less stable with less G . C pairs...

  18. The nicotinic acetylcholine receptor gene family of the silkworm, Bombyx mori

    Directory of Open Access Journals (Sweden)

    Zhang Chuan-Xi

    2007-09-01

    Full Text Available Abstract Background Nicotinic acetylcholine receptors (nAChRs mediate fast synaptic cholinergic transmission in the insect central nervous system. The insect nAChR is the molecular target of a class of insecticides, neonicotinoids. Like mammalian nAChRs, insect nAChRs are considered to be made up of five subunits, coded by homologous genes belonging to the same family. The nAChR subunit genes of Drosophila melanogaster, Apis mellifera and Anopheles gambiae have been cloned previously based on their genome sequences. The silkworm Bombyx mori is a model insect of Lepidoptera, among which are many agricultural pests. Identification and characterization of B. mori nAChR genes could provide valuable basic information for this important family of receptor genes and for the study of the molecular mechanisms of neonicotinoid action and resistance. Results We searched the genome sequence database of B. mori with the fruit fly and honeybee nAChRs by tBlastn and cloned all putative silkworm nAChR cDNAs by reverse transcriptase-polymerase chain reaction (RT-PCR and rapid amplification of cDNA ends (RACE methods. B. mori appears to have the largest known insect nAChR gene family to date, including nine α-type subunits and three β-type subunits. The silkworm possesses three genes having low identity with others, including one α and two β subunits, α9, β2 and β3. Like the fruit fly and honeybee counterparts, silkworm nAChR gene α6 has RNA-editing sites, and α4, α6 and α8 undergo alternative splicing. In particular, alternative exon 7 of Bmα8 may have arisen from a recent duplication event. Truncated transcripts were found for Bmα4 and Bmα5. Conclusion B. mori possesses a largest known insect nAChR gene family characterized to date, including nine α-type subunits and three β-type subunits. RNA-editing, alternative splicing and truncated transcripts were found in several subunit genes, which might enhance the diversity of the gene family.

  19. Binding specificity of Bacillus thuringiensis Cry1Aa for purified, native Bombyx mori aminopeptidase N and cadherin-like receptors

    Directory of Open Access Journals (Sweden)

    Jenkins Jeremy L

    2001-10-01

    Full Text Available Abstract Background To better understand the molecular interactions of Bt toxins with non-target insects, we have examined the real-time binding specificity and affinity of Cry1 toxins to native silkworm (Bombyx mori midgut receptors. Previous studies on B. mori receptors utilized brush border membrane vesicles or purifed receptors in blot-type assays. Results The Bombyx mori (silkworm aminopeptidase N (APN and cadherin-like receptors for Bacillus thuringiensis insecticidal Cry1Aa toxin were purified and their real-time binding affinities for Cry toxins were examined by surface plasmon resonance. Cry1Ab and Cry1Ac toxins did not bind to the immobilized native receptors, correlating with their low toxicities. Cry1Aa displayed moderate affinity for B. mori APN (75 nM, and unusually tight binding to the cadherin-like receptor (2.6 nM, which results from slow dissociation rates. The binding of a hybrid toxin (Aa/Aa/Ac was identical to Cry1Aa. Conclusions These results indicate domain II of Cry1Aa is essential for binding to native B. mori receptors and for toxicity. Moreover, the high-affinity binding of Cry1Aa to native cadherin-like receptor emphasizes the importance of this receptor class for Bt toxin research.

  20. Genome-wide transcriptional response of silkworm (Bombyx mori to infection by the microsporidian Nosema bombycis.

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    Zhengang Ma

    Full Text Available Microsporidia have attracted much attention because they infect a variety of species ranging from protists to mammals, including immunocompromised patients with AIDS or cancer. Aside from the study on Nosema ceranae, few works have focused on elucidating the mechanism in host response to microsporidia infection. Nosema bombycis is a pathogen of silkworm pébrine that causes great economic losses to the silkworm industry. Detailed understanding of the host (Bombyx mori response to infection by N. bombycis is helpful for prevention of this disease. A genome-wide survey of the gene expression profile at 2, 4, 6 and 8 days post-infection by N. bombycis was performed and results showed that 64, 244, 1,328, 1,887 genes were induced, respectively. Up to 124 genes, which are involved in basal metabolism pathways, were modulated. Notably, B. mori genes that play a role in juvenile hormone synthesis and metabolism pathways were induced, suggesting that the host may accumulate JH as a response to infection. Interestingly, N. bombycis can inhibit the silkworm serine protease cascade melanization pathway in hemolymph, which may be due to the secretion of serpins in the microsporidia. N. bombycis also induced up-regulation of several cellular immune factors, in which CTL11 has been suggested to be involved in both spore recognition and immune signal transduction. Microarray and real-time PCR analysis indicated the activation of silkworm Toll and JAK/STAT pathways. The notable up-regulation of antimicrobial peptides, including gloverins, lebocins and moricins, strongly indicated that antimicrobial peptide defense mechanisms were triggered to resist the invasive microsporidia. An analysis of N. bombycis-specific response factors suggested their important roles in anti-microsporidia defense. Overall, this study primarily provides insight into the potential molecular mechanisms for the host-parasite interaction between B. mori and N. bombycis and may provide a

  1. Catalase from the silkworm, Bombyx mori: gene sequence, distribution, and overexpression.

    Science.gov (United States)

    Yamamoto, Kohji; Banno, Yutaka; Fujii, Hiroshi; Miake, Fumio; Kashige, Nobuhiro; Aso, Yoichi

    2005-04-01

    Living organisms require mechanisms regulating reactive oxygen species (ROS) such as hydrogen peroxide and superoxide anion. Catalase is one of the regulatory enzymes and facilitates the degradation of hydrogen peroxide to oxygen and water. Biochemical information on an insect catalase is, however, insufficient. Using mRNA from fat body of the silkworm, Bombyx mori, a cDNA encoding a putative catalase was amplified by reverse transcriptase-polymerase chain reaction and sequenced. The deduced amino acid sequence comprised 507 residues with more than seventy residues forming a scaffold for a heme cofactor conserved. The sequence showed 71% and 66% identities to those of the Drosophila melanogaster and Apis mellifera catalases, respectively; the catalase from B. mori was estimated to be phylogenetically close to that from A. mellifera. The transcripts of the gene and the catalase activity were distributed in diverse tissues of B. mori, suggesting its ubiquitous nature. Using the gene, a recombinant catalase (rCAT) was functionally overexpressed in a soluble form using Escherichia coli, purified to homogeneity, and characterized. The pH-optimum of rCAT was broad around pH 8.0. More than 80% of the original rCAT activity was retained after incubation in the following conditions: at pH 8-11 and 4 degrees C for 24 h; at pH 7 and temperatures below 50 degrees C for 30 min. The Michaelis constant for hydrogen peroxide was evaluated to be 28 mM at pH 6.5 and 30 degrees C. rCAT was suggested to be a member of the typical catalase family. PMID:15763464

  2. Functional expression of a Bombyx mori cocoonase: potential application for silk degumming

    Institute of Scientific and Technical Information of China (English)

    Prangprapai Rodbumrer; Dumrongkiet Arthan; Utai Uyen; Jirundon Yuvaniyama; Jisnuson Svasti; Pramvadee Y.Wongsaengchantra

    2012-01-01

    Cocoon,a shelter for larva development to silk moth,contains the fibrous protein fibroin,which is coated by the globular protein sericin.Emergence of the silk moth requires the action of cocoonase,a protease secreted by the pupa.The full-length prococoonase cDNA,with 780 bp open reading frame encoding 260 amino acids,was cloned by reverse transcription from total RNA of the head of 6-day-old Thai-silk Bombyx mori pupa.Only the gene fragment lacking the propeptide encoding sequence was successfully expressed in Pichia pastoris,yielding an extracellularly active cocoonase.The recombinant cocoonase was purified to homogeneity by 80% ammonium-suffate fractionation and CM-Sepharose chromatography,and its internal peptide sequences were analyzed by nano liquid chromatographymass spectrometry/mass spectrometry.This monomeric protein has native molecular weight of 26 kDa by gel exclusion analysis and 25 kDa subunit size by sodium dodecyl sulphate-polyacrylamide gel electrophoresis.The enzyme hydrolyses sericin but does not hydrolyse fibroin,as shown by radial diffusion on thin-layer enzyme assay (RD-TEA).Scanning electron microscopy showed that purified recombinant cocoonase could remove sericin from natural silk completely in 24 h,without damaging fibroin,using only 1immobilized sericin unit (ISU) of enzyme as determined by RD-TEA.Natural cocoonase isolated from B.mori pupa could also digest sericin effectively,but required more enzymes (2 ISU) and longer time (48 h).In comparison,a commercial enzyme,alcalase,with the same activity not only showed less complete digestion of sericin but also caused damage of fibroin.These results suggest that recombinant B.mori cocoonase is potentially useful for silk degumming.

  3. Orthologs of Human Disease Associated Genes and RNAi Analysis of Silencing Insulin Receptor Gene in Bombyx mori

    Directory of Open Access Journals (Sweden)

    Zan Zhang

    2014-10-01

    Full Text Available The silkworm, Bombyx mori L., is an important economic insect that has been domesticated for thousands of years to produce silk. It is our great interest to investigate the possibility of developing the B. mori as human disease model. We searched the orthologs of human disease associated genes in the B. mori by bi-directional best hits of BLAST and confirmed by searching the OrthoDB. In total, 5006 genes corresponding to 1612 kinds of human diseases had orthologs in the B. mori, among which, there are 25 genes associated with diabetes mellitus. Of these, we selected the insulin receptor gene of the B. mori (Bm-INSR to study its expression in different tissues and at different developmental stages and tissues. Quantitative PCR showed that Bm-INSR was highly expressed in the Malpighian tubules but expressed at low levels in the testis. It was highly expressed in the 3rd and 4th instar larvae, and adult. We knocked down Bm-INSR expression using RNA interference. The abundance of Bm-INSR transcripts were dramatically reduced to ~4% of the control level at 6 days after dsRNA injection and the RNAi-treated B. mori individuals showed apparent growth inhibition and malformation such as abnormal body color in black, which is the typical symptom of diabetic patients. Our results demonstrate that B. mori has potential use as an animal model for diabetic mellitus research.

  4. A novel angiotensin-І converting enzyme (ACE) inhibitory peptide from gastrointestinal protease hydrolysate of silkworm pupa (Bombyx mori) protein: Biochemical characterization and molecular docking study.

    Science.gov (United States)

    Wu, Qiongying; Jia, Junqiang; Yan, Hui; Du, Jinjuan; Gui, Zhongzheng

    2015-06-01

    Silkworm pupa (Bombyx mori) protein was hydrolyzed using gastrointestinal endopeptidases (pepsin, trypsin and α-chymotrypsin). Then, the hydrolysate was purified sequentially by ultrafiltration, gel filtration chromatography and RP-HPLC. A novel ACE inhibitory peptide, Ala-Ser-Leu, with the IC50 value of 102.15μM, was identified by IT-MS/MS. This is the first report of Ala-Ser-Leu from natural protein. Lineweaver-Burk plots suggest that the peptide is a competitive inhibitor against ACE. The molecular docking studies revealed that the ACE inhibition of Ala-Ser-Leu is mainly attributed to forming very strong hydrogen bonds with the S1 pocket (Ala354) and the S2 pocket (Gln281 and His353). The results indicate that silkworm pupa (B. mori) protein or its gastrointestinal protease hydrolysate could be used as a functional ingredient in auxiliary therapeutic foods against hypertension. PMID:25111373

  5. Scientific validation of cardioprotective attribute by standardized extract of Bombyx mori against doxorubicin-induced cardiotoxicity in murine model

    OpenAIRE

    Khan, Masood S.; Mhaveer Singh, Mhaveer; Khan, Mohammad A.; Arya, D. S.; Ahmad, Sayeed

    2014-01-01

    Doxorubicin (DOX) is an excellent antineoplastic agent used for the treatment of hematological and solid malignancies. The aqueous extract of Bombyx mori (BMAE) contains amino acids and some flavonoids with obvious cardioprot ective effect. The aim of this study was to investigate the possible protective effect of BMAE against DOX-induced cardiotoxicity and its underlying mechanisms on murine model. The metabolic profiling of BMAE was carried out by Ultra Performance Liquid Chromatography-Mas...

  6. Isolation and Characterization of Lipase-Producing Bacteria in the Intestine of the Silkworm, Bombyx mori, Reared on Different Forage

    OpenAIRE

    Feng, Wei; Wang, Xiao-Qiang; Wei ZHOU; Liu, Guang-ying; Wan, Yong-Ji

    2011-01-01

    The silkworm, Bombyx mori L. (Lepidoptera: Bombycidae), an oligophagous insect that mainly feeds on mulberry leaves, is susceptible to entomopathogen infection when reared with tricuspid cudrania leaves. A total of 56 dominant bacterial strains, classified into 12 phylotypes based on bacteriological properties and analysis of 16S rRNA genes, were isolated from the intestine of the fourth and fifth instar silkworm larvae. Ten and seven phylotypes exist in the intestine of the silkworm larvae r...

  7. Fibroin and Sericin from Bombyx mori Silk Stimulate Cell Migration through Upregulation and Phosphorylation of c-Jun

    OpenAIRE

    Celia Martínez-Mora; Anna Mrowiec; Eva María García-Vizcaíno; Antonia Alcaraz; José Luis Cenis; Francisco José Nicolás

    2012-01-01

    Wound healing is a biological process directed to the restoration of tissue that has suffered an injury. An important phase of wound healing is the generation of a basal epithelium able to wholly replace the epidermis of the wound. A broad range of products derived from fibroin and sericin from Bombyx mori silk are used to stimulate wound healing. However, so far the molecular mechanism underlying this phenomenon has not been elucidated. The aim of this work was to determine the molecular bas...

  8. Hox transcription factor Antp regulates sericin-1 gene expression in the terminal differentiated silk gland of Bombyx mori

    OpenAIRE

    Kimoto, Mai; Tsubota, Takuya; Uchino, Keiro; Sezutsu, Hideki; Takiya, Shigeharu

    2014-01-01

    Hox genes are well-known master regulators in developmental morphogenesis along the anteroposterior axis of animals. However, the molecular mechanisms by which Hox proteins regulate their target genes and determine cell fates are not fully understood. The silk gland of Bombyx mori is a tubular tissue divided into several subparts along the anteroposterior axis, and the silk genes are expressed with specific patterns. The sericin-1 gene (ser1) is expressed in the middle silk gland (MSG) with s...

  9. Influence of sericin in alleviating the hydrogen peroxide induced oxidative stress in silkworm Bombyx mori: role of the amino acids

    OpenAIRE

    AS Micheal; Subramanyam, M

    2014-01-01

    Sericin is an important peptide derived from silk fibre spun by the silkworm Bombyx mori and has various biological activities. The aim of the present study was to characterize the major constituents of sericin that are providing cytoprotective effect against hydrogen peroxide-induced cell damage in midgut epithelial cells and hemocytes of silkworm. Extracted sericin was subjected to LCMS analysis for amino acid composition. Isolated cells of midgut and hemocytes were incubated with sericin o...

  10. Effect of vertebrate hormones and prostaglandins on growth, silk quality and metabolic activities of Bombyx mori L

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    The economic importance of silkworm has moved biologists to explore various intricate mechanisms of the action of vertebrate hormones. The dietary administration of several vertebrate hormones and prostaglandins enhanced both developmental and metabolic processes of silkworm, Bombyx mori L. The main objective of sericulture research is to apply the results to achieve superior quality silk and greater output, to apply lab findings to achieve desirable ecenomic results.

  11. Direct and indirect effects of RNA interference against pyridoxal kinase and pyridoxine 5'-phosphate oxidase genes in Bombyx mori.

    Science.gov (United States)

    Huang, ShuoHao; Yao, LiLi; Zhang, JianYun; Huang, LongQuan

    2016-08-01

    Vitamin B6 comprises six interconvertible pyridine compounds (vitamers), among which pyridoxal 5'-phosphate is a coenzyme involved in a high diversity of biochemical reactions. Humans and animals obtain B6 vitamers from diet, and synthesize pyridoxal 5'-phosphate by pyridoxal kinase and pyridoxine 5'-phosphate oxidase. Currently, little is known on how pyridoxal 5'-phosphate biosynthesis is regulated, and pyridoxal 5'-phosphate is supplied to meet their requirement in terms of cofactor. Bombyx mori is a large silk-secreting insect, in which protein metabolism is most active, and the vitamin B6 demand is high. In this study, we successfully down-regulated the gene expression of pyridoxal kinase and pyridoxine 5'-phosphate oxidase by body cavity injection of synthesized double-stranded small interfering RNA to 5th instar larvae of Bombyx mori, and analyzed the gene transcription levels of pyridoxal 5'-phosphate dependent enzymes, phosphoserine aminotransferase and glutamic-oxaloacetic transaminase. Results show that the gene expression of pyridoxal kinase and pyridoxine 5'-phosphate oxidase has a greater impact on the gene transcription of enzymes using pyridoxal 5'-phosphate as a cofactor in Bombyx mori. Our study suggests that pyridoxal 5'-phosphate biosynthesis and dynamic balance may be regulated by genetic networks. PMID:27106120

  12. Transcriptomic Analysis of Resistant and Susceptible Bombyx mori Strains Following BmNPV Infection Provides Insights into the Antiviral Mechanisms.

    Science.gov (United States)

    Li, Gang; Qian, Heying; Luo, Xufang; Xu, Pingzhen; Yang, Jianhua; Liu, Mingzhu; Xu, Anying

    2016-01-01

    Purpose. To decipher transcriptomic changes and related genes with potential functions against Bombyx mori nucleopolyhedrovirus infection and to increase the understanding of the enhanced virus resistance of silkworm on the transcriptomic level. Methods. We assembled and annotated transcriptomes of the Qiufeng (susceptible to infection) and QiufengN (resistant to infection) strains and performed comparative analysis in order to decipher transcriptomic changes and related genes with potential functions against BmNPV infection. Results. A total of 78,408 SNPs were identified in the Qiufeng strain of silkworm and 56,786 SNPs were identified in QiufengN strain. Besides, novel AS events were found in these 2 strains. In addition, 1,728 DEGs were identified in the QiufengN strain compared with Qiufeng strain. These DEGs were involved in GO terms related to membrane, metabolism, binding and catalytic activity, cellular processes, and organismal systems. The highest levels of gene representation were found in oxidative phosphorylation, phagosome, TCA cycle, arginine and proline metabolism, and pyruvate metabolism. Additionally, COG analysis indicated that DEGs were involved in "amino acid transport and metabolism" and "carbohydrate transport and metabolism." Conclusion. We identified a series of major pathological changes in silkworm following infection and several functions were related to the antiviral mechanisms of silkworm. PMID:27195279

  13. Juvenile Hormone Analogues, Methoprene and Fenoxycarb Dose-Dependently Enhance Certain Enzyme Activities in the Silkworm Bombyx Mori (L

    Directory of Open Access Journals (Sweden)

    M. Rajeswara Rao

    2008-06-01

    Full Text Available Use of Juvenile Hormone Analogues (JHA in sericulture practices has been shown to boost good cocoon yield; their effect has been determined to be dose-dependent. We studied the impact of low doses of JHA compounds such as methoprene and fenoxycarb on selected key enzymatic activities of the silkworm Bombyx mori. Methoprene and fenoxycarb at doses of 1.0 μg and 3.0fg/larvae/48 hours showed enhancement of the 5th instar B. mori larval muscle and silkgland protease, aspartate aminotransaminase (AAT and alanine aminotransaminase (ALAT, adenosine triphosphate synthase (ATPase and cytochrome-c-oxidase (CCO activity levels, indicating an upsurge in the overall oxidative metabolism of the B.mori larval tissues.

  14. Identification and Characterization of Novel Chitin-Binding Proteins from the Larval Cuticle of Silkworm, Bombyx mori.

    Science.gov (United States)

    Dong, Zhaoming; Zhang, Weiwei; Zhang, Yan; Zhang, Xiaolu; Zhao, Ping; Xia, Qingyou

    2016-05-01

    Cuticle is mainly made of chitin filaments embedded in a matrix of cuticular proteins (CPs). Cuticular chitins have minor differences, whereas CPs are widely variable with respect to their sequences and structures. To understand the molecular basis underlying the mechanical properties of cuticle, it is necessary to know which CPs interact with chitin and how they are assembled into the cuticle structure. In the present study, a chitin-binding assay was performed followed by liquid chromatography-tandem mass spectrometry to identify the extracted proteins from the larval cuticle of silkworm, Bombyx mori. There were 463 proteins identified from the silkworm larval cuticle, 200 of which were recovered in the chitin-binding fraction. A total of 103 proteins were annotated as CPs, which were classified into 11 CP families based on their conserved motifs, including CPR, CPAP, CPT, CPF and CPFL, CPCFC, chitin_bind 3, BmCPH2 homologues, BmCPH9 homologues, BmCPG1 homologues, BmCPG20 homologues, and BmCPG21 homologues. A total of five CP families were newly identified in the chitin-binding fraction, thereby providing new information and insight into the composition, structure, and function of the silkworm larval cuticle. PMID:26972338

  15. Extensive conserved synteny of genes between the karyotypes of Manduca sexta and Bombyx mori revealed by BAC-FISH mapping.

    Directory of Open Access Journals (Sweden)

    Yuji Yasukochi

    Full Text Available BACKGROUND: Genome sequencing projects have been completed for several species representing four highly diverged holometabolous insect orders, Diptera, Hymenoptera, Coleoptera, and Lepidoptera. The striking evolutionary diversity of insects argues a need for efficient methods to apply genome information from such models to genetically uncharacterized species. Constructing conserved synteny maps plays a crucial role in this task. Here, we demonstrate the use of fluorescence in situ hybridization with bacterial artificial chromosome probes as a powerful tool for physical mapping of genes and comparative genome analysis in Lepidoptera, which have numerous and morphologically uniform holokinetic chromosomes. METHODOLOGY/PRINCIPAL FINDINGS: We isolated 214 clones containing 159 orthologs of well conserved single-copy genes of a sequenced lepidopteran model, the silkworm, Bombyx mori, from a BAC library of a sphingid with an unexplored genome, the tobacco hornworm, Manduca sexta. We then constructed a BAC-FISH karyotype identifying all 28 chromosomes of M. sexta by mapping 124 loci using the corresponding BAC clones. BAC probes from three M. sexta chromosomes also generated clear signals on the corresponding chromosomes of the convolvulus hawk moth, Agrius convolvuli, which belongs to the same subfamily, Sphinginae, as M. sexta. CONCLUSIONS/SIGNIFICANCE: Comparison of the M. sexta BAC physical map with the linkage map and genome sequence of B. mori pointed to extensive conserved synteny including conserved gene order in most chromosomes. Only a few rearrangements, including three inversions, three translocations, and two fission/fusion events were estimated to have occurred after the divergence of Bombycidae and Sphingidae. These results add to accumulating evidence for the stability of lepidopteran genomes. Generating signals on A. convolvuli chromosomes using heterologous M. sexta probes demonstrated that BAC-FISH with orthologous sequences can be

  16. Molecular cloning and function of ecdysis-triggering hormones in the silkworm Bombyx mori.

    Science.gov (United States)

    Zitnan, Dusan; Hollar, Laura; Spalovská, Ivana; Takác, Peter; Zitnanová, Inka; Gill, Sarjeet S; Adams, Michael E

    2002-11-01

    Inka cells of the epitracheal endocrine system produce peptide hormones involved in the regulation of insect ecdysis. In the silkworm Bombyx mori, injection of Inka cell extract into pharate larvae, pupae or adults activates the ecdysis behavioural sequence. In the present study, we report the identification of three peptides in these extracts, pre-ecdysis-triggering hormone (PETH), ecdysis-triggering hormone (ETH) and ETH-associated peptide (ETH-AP), which are encoded by the same cDNA precursor. Strong immunoreactivity associated with each peptide in Inka cells prior to ecdysis disappears during each ecdysis, indicating complete release of these peptides. Injection of either PETH or ETH alone is sufficient to elicit the entire ecdysis behavioural sequence through the direct action on abdominal ganglia; cephalic and thoracic ganglia are not required for the transition from pre-ecdysis to ecdysis behaviour. Our in vitro data provide evidence that these peptides control the entire ecdysis behavioural sequence through activation of specific circuits in the nervous system. Ecdysis of intact larvae is associated with the central release of eclosion hormone (EH) and elevation of cyclic 3',5'-guanosine monophosphate (cGMP) in the ventral nerve cord. However, injection of ETH into isolated abdomens induces cGMP elevation and ecdysis behaviour without a detectable release of EH, suggesting that an additional central factor(s) may be involved in the activation of this process. Our findings provide the first detailed account of the natural and hormonally induced behavioural sequence preceding larval, pupal and adult ecdyses of B. mori and highlight significant differences in the neuro-endocrine activation of pre-ecdysis and ecdysis behaviours compared with the related moth, Manduca sexta. PMID:12364399

  17. Regeneration of Bombyx mori silk nanofibers and nanocomposite fibrils by the electrospinning process

    Science.gov (United States)

    Ayutsede, Jonathan Eyitouyo

    In recent years, there has been significant interest in the utilization of natural materials for novel nanoproducts such as tissue engineered scaffolds. Silkworm silk fibers represent one of the strongest natural fibers known. Silkworm silk, a protein-based natural biopolymer, has received renewed interest in recent years due to its unique properties (strength, toughness) and potential applications such as smart textiles, protective clothing and tissue engineering. The traditional 10--20 mum diameter, triangular-shaped Bombyx mori fibers have remained unchanged over the years. However, in our study, we examine the scientific implication and potential applications of reducing the diameter to the nanoscale, changing the triangular shape of the fiber and adding nanofillers in the form of single wall carbon nanotubes (SWNT) by the electrospinning process. The electrospinning process preserves the natural conformation of the silk (random and beta-sheet). The feasibility of changing the properties of the electrospun nanofibers by post processing treatments (annealing and chemical treatment) was investigated. B. mori silk fibroin solution (formic acid) was successfully electrospun to produce uniform nanofibers (as small as 12 nm). Response Surface Methodology (RSM) was applied for the first time to experimental results of electrospinning, to develop a processing window that can reproduce regenerated silk nanofibers of a predictable size (d nature of the silk-SWNT interactions. A new visualization system was developed to characterize the transport properties of the nanofibrous assemblies. The morphological, chemical, structural and mechanical properties of the nanofibers were determined by field emission environmental scanning microscopy, Fourier transform infrared and Raman spectroscopy, wide angle x-ray diffraction and microtensile tester respectively.

  18. Surface modification and properties of Bombyx mori silk fibroin films by antimicrobial peptide

    Science.gov (United States)

    Bai, Liqiang; Zhu, Liangjun; Min, Sijia; Liu, Lin; Cai, Yurong; Yao, Juming

    2008-03-01

    The Bombyx mori silk fibroin films (SFFs) were modified by a Cecropin B ( CB) antimicrobial peptide, (NH 2)-NGIVKAGPAIAVLGEAAL-CONH 2, using the carbodiimide chemistry method. In order to avoid the dissolution of films during the modification procedure, the SFFs were first treated with 60% (v/v) ethanol aqueous solution, resulting a structural transition from unstable silk I to silk II. The investigation of modification conditions showed that the surface-modified SFFs had the satisfied antimicrobial activity and durability when they were activated by EDC·HCl/NHS solution followed by a treatment in CB peptide/PBS buffer (pH 6.5 or 8) solution at ambient temperature for 2 h. Moreover, the surface-modified SFFs showed the smaller contact angle due to the hydrophilic antimicrobial peptides coupled on the film surface, which is essential for the cell adhesion and proliferation. AFM results indicated that the surface roughness of SFFs was considerably increased after the modification by the peptides. The elemental composition analysis results also suggested that the peptides were tightly coupled to the surface of SFFs. This approach may provide a new option to engineer the surface-modified implanted materials preventing the biomaterial-centered infection (BCI).

  19. Surface modification and properties of Bombyx mori silk fibroin films by antimicrobial peptide

    International Nuclear Information System (INIS)

    The Bombyx mori silk fibroin films (SFFs) were modified by a Cecropin B (CB) antimicrobial peptide, (NH2)-NGIVKAGPAIAVLGEAAL-CONH2, using the carbodiimide chemistry method. In order to avoid the dissolution of films during the modification procedure, the SFFs were first treated with 60% (v/v) ethanol aqueous solution, resulting a structural transition from unstable silk I to silk II. The investigation of modification conditions showed that the surface-modified SFFs had the satisfied antimicrobial activity and durability when they were activated by EDC.HCl/NHS solution followed by a treatment in CB peptide/PBS buffer (pH 6.5 or 8) solution at ambient temperature for 2 h. Moreover, the surface-modified SFFs showed the smaller contact angle due to the hydrophilic antimicrobial peptides coupled on the film surface, which is essential for the cell adhesion and proliferation. AFM results indicated that the surface roughness of SFFs was considerably increased after the modification by the peptides. The elemental composition analysis results also suggested that the peptides were tightly coupled to the surface of SFFs. This approach may provide a new option to engineer the surface-modified implanted materials preventing the biomaterial-centered infection (BCI)

  20. Appearance of differentiated cells derived from polar body nuclei in the silkworm, Bombyx mori

    Directory of Open Access Journals (Sweden)

    Hiroki eSakai

    2013-09-01

    Full Text Available AbstractIn Bombyx mori, polar body nuclei are observed until 9h after egg lying, however, the fate of polar body nuclei remains unclear. To examine the fate of polar body nuclei, we employed a mutation of serosal cell pigmentation, pink-eyed white egg (pe. The heterozygous pe/+pe females produced black serosal cells in white eggs, while pe/pe females did not produce black serosal cells in white eggs. These results suggest that the appearance of black serosal cells in white eggs depends on the genotype (pe/ +pe of the mother. Because the polar body nuclei had +pe genes in the white eggs laid by a pe/ +pe female, polar body nuclei participate in development and differentiate into functional cell (serosal cells. Analyses of serosal cells pigmentation indicated that approximately 30% of the eggs contained polar-body-nucleus-derived cells. These results demonstrate that polar-body-nucleus-derived cells appeared at a high frequency under natural conditions. Approximately 80% of polar-body-nucleus-derived cells appeared near the anterior pole and the dorsal side, which is opposite to where embryogenesis occurs. The number of cells derived from the polar body nuclei was very low. Approximately 26 % of these eggs contained only one black serosal cell. PCR-based analysis revealed that the polar-body-nucleus-derived cells disappeared in late embryonic stages (stage 25. Overall, polar-body-nuclei-derived cells were unlikely to contribute to embryos.

  1. Efficient silkworm expression of human GPCR (nociceptin receptor) by a Bombyx mori bacmid DNA system

    International Nuclear Information System (INIS)

    Guanine nucleotide-binding protein (G protein) coupled receptors (GPCRs) are frequently expressed by a baculovirus expression vector system (BEVS). We recently established a novel BEVS using the bacmid system of Bombyx mori nucleopolyhedrovirus (BmNPV), which is directly applicable for protein expression in silkworms. Here, we report the first example of GPCR expression in silkworms by the simple injection of BmNPV bacmid DNA. Human nociceptin receptor, an inhibitory GPCR, and its fusion protein with inhibitory G protein alpha subunit (Giα) were both successfully expressed in the fat bodies of silkworm larvae as well as in the BmNPV viral fraction. Its yield was much higher than that from Sf9 cells. The microsomal fractions including the nociceptin receptor fusion, which are easily prepared by only centrifugation steps, exhibited [35S]GTPγS-binding activity upon specific stimulation by nociceptin. Therefore, this rapid method is easy-to-use and has a high expression level, and thus will be an important tool for human GPCR production.

  2. Expression pattern of enzymes related to juvenile hormone metabolism in the silkworm, Bombyx mori L.

    Science.gov (United States)

    Hua-Jun, Yang; Fang, Zhou; Awquib, Sabhat; Malik, Firdose Ahmad; Roy, Bhaskar; Xing-Hua, Li; Jia-Biao, Hu; Chun-Guang, Sun; Niu, Yan-Shan; Yun-Gen, Miao

    2011-10-01

    The physiological balance of juvenile hormone (JH) in insects depends on its biosynthesis and degradation pathway. Three key enzymes namely, juvenile hormone esterase (JHE), juvenile hormone epoxide hydrolase (JHEH) and juvenile hormone diol kinase (JHDK) are required for degradation in insects. Our present results showed that JHE and JHEH exhibited expression in almost all the tissues. This indicated that JHE and JHEH might degrade JH simultaneously. In addition, the highest levels of JHDK were observed in the midgut, with trace level being found in the malpighian tubule and haemocytes. Since the midgut is a digestive organ and not a JH target, it was hypothesized that both JHE and JHEH hydrolyzed JH to JH diol (JHd) which was then transported to midgut and hydrolyzed further by JHDK, to be finally excreted out of the body. Also the expression studies on JH degradation enzymes in different tissues and stages indicated that the activities of the three enzymes are specific and coincident with the JH functions in silkworm, Bombyx mori L. PMID:21107706

  3. Molecular cloning, expression and characterization of acylpeptide hydrolase in the silkworm, Bombyx mori.

    Science.gov (United States)

    Fu, Ping; Sun, Wei; Zhang, Ze

    2016-04-10

    Acylpeptide hydrolase (APH) can catalyze the release of the N-terminal amino acid from acetylated peptides. There were many documented examples of this enzyme in various prokaryotic and eukaryotic organisms. However, knowledge about APH in insects still remains unknown. In this study, we cloned and sequenced a putative silkworm Bombyx mori APH (BmAPH) gene. The BmAPH gene encodes a protein of 710 amino acids with a predicted molecular mass of 78.5kDa. The putative BmAPH and mammal APHs share about 36% amino acid sequence identity, yet key catalytic residues are conserved (Ser566, Asp654, and His686). Expression and purification of the recombinant BmAPH in Escherichia coli showed that it has acylpeptide hydrolase activity toward the traditional substrate, Ac-Ala-pNA. Furthermore, organophosphorus (OP) insecticides, chlorpyrifos, phoxim, and malathion, significantly inhibited the activity of the APH both in vitro and in vivo. In addition, BmAPH was expressed in all tested tissues and developmental stages of the silkworm. Finally, immunohistochemistry analysis showed that BmAPH protein was localized in the basement membranes. These results suggested that BmAPH may be involved in enhancing silkworm tolerance to the OP insecticides. In a word, our results provide evidence for understanding of the biological function of APH in insects. PMID:26778207

  4. Intestinal microecology associated with fluoride resistance capability of the silkworm (Bombyx mori L.).

    Science.gov (United States)

    Li, Guan-Nan; Xia, Xue-Juan; Tang, Wen-Chao; Zhu, Yong

    2016-08-01

    The silkworm (Bombyx mori L.) is an ideal model of Lepidoptera. However, the diversity and function of the intestinal microbiota in the gut of silkworm remain largely unknown. Changes in the intestinal microecology in fluoride-resistant strain T6 and fluoride-susceptible strain 734 of the silkworm in response to fluoride exposure were investigated. T6 and 734 were treated with 200 mg/kg fluoride (designated as T6-T and 734-T groups) and deionized water (designated as T6-C and 734-C groups). Culture-dependent approach revealed that the numbers of intestinal bacteria in the 734-T group significantly decreased compared with that in the 734-C group (4.8 ± 0.6 × 10(7) CFU/mL vs. 7.5 ± 0.7 × 10(7) CFU/mL; P diverse than those in the other groups. The bacterial community was composed of two dominant groups (Firmicutes and Proteobacteria). Principal component analyses revealed a significant difference in the composition of the intestinal microbiota in the 734-T group compared with those in the other groups. Thaumarchaeota and Euryarchaeota were more abundant in the 734-T group, but they were less abundant in the other groups. This study enhances our understanding about the diversity and function of silkworm intestinal microbiota in response to fluoride exposure among silkworm strains with diverse resistance. PMID:27147533

  5. Preparation and characterization of regenerated fiber from the aqueous solution of Bombyx mori cocoon silk fibroin

    Energy Technology Data Exchange (ETDEWEB)

    Zhu Zhenghua [Department of Biotechnology, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588 (Japan); Department of Application Engineering, ZheJiang Vocational College of Economic and Trade, HangZhou, ZheJiang 310018 (China); Imada, Takuzo [Department of Biotechnology, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588 (Japan); Asakura, Tetsuo, E-mail: asakura@cc.tuat.ac.jp [Department of Biotechnology, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588 (Japan)

    2009-10-15

    The regenerated silk fibers with high strength and high biodegradability were prepared from the aqueous solution of Bombyx mori silk fibroin from cocoons with wet spinning method. Although the tensile strength of the regenerated silk fibroin fiber, 210 MPa is still half of the strength of native silk fiber, the diameter of the fiber is about 100 {mu}m which is suitable for monofilament of suture together with high biodegradability. The high concentration (30%, w/v) of the aqueous solution of the silk fibroin which corresponds to the high concentration in the middle silkgland of silkworm was obtained. This was performed by adjusting the pH of the aqueous solution to 10.4 which corresponds to pK{sub a} value of the OH group of Tyr residues in the silk fibroin. The mixed solvent, methanol/acetic acid (7:3 in volume ratio) was used as coagulant solvent for preparing the regenerated fiber. The structural change of silk fibroin fiber by stretching was monitored with both {sup 13}C solid state NMR and X-ray diffraction methods, indicating that the high strength of the fiber is related with the long-range orientation of the silk fibroin chain with {beta}-sheet structure.

  6. Preparation and characterization of regenerated fiber from the aqueous solution of Bombyx mori cocoon silk fibroin

    International Nuclear Information System (INIS)

    The regenerated silk fibers with high strength and high biodegradability were prepared from the aqueous solution of Bombyx mori silk fibroin from cocoons with wet spinning method. Although the tensile strength of the regenerated silk fibroin fiber, 210 MPa is still half of the strength of native silk fiber, the diameter of the fiber is about 100 μm which is suitable for monofilament of suture together with high biodegradability. The high concentration (30%, w/v) of the aqueous solution of the silk fibroin which corresponds to the high concentration in the middle silkgland of silkworm was obtained. This was performed by adjusting the pH of the aqueous solution to 10.4 which corresponds to pKa value of the OH group of Tyr residues in the silk fibroin. The mixed solvent, methanol/acetic acid (7:3 in volume ratio) was used as coagulant solvent for preparing the regenerated fiber. The structural change of silk fibroin fiber by stretching was monitored with both 13C solid state NMR and X-ray diffraction methods, indicating that the high strength of the fiber is related with the long-range orientation of the silk fibroin chain with β-sheet structure.

  7. Structural Analysis of Fibroin Heavy Chain Signal Peptide of Silkworm Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    Sheng-Peng WANG; Ting-Qing GUO; Xiu-Yang GUO; Jun-Ting HUANG; Chang-De LU

    2006-01-01

    To study the minimal length required for the secretion of recombinant proteins and silk proteins in posterior silk gland, the signal peptide (SP) of the fibroin heavy chain (FibH) of silkworm Bombyx mori was systematically shortened from the C-terminal. Its effect on the secretion of protein was observed using enhanced green fluorescent protein (EGFP) as a reporter. Secretion of EGFP fusion proteins was examined under fluorescence microscope. FibH SPs with lengths of 20, 18, 16 and 12 a.a. can direct the secretion of the reporter, yet those with lengths of 11, 10, 9, 8 and 1 a.a. can not. When the FibH SP was shortened to 12 a.a., the secretion efficiency was decreased slightly and cleavage occurred within EGFP.When 16 a.a. of the FibH SP were used, the secretion of fusion protein was normal and the cleavage site was between the Gly-Ser linker and Met, the starting amino acid of EGFP. These findings are applicable for the expression of foreign proteins in silkworm silk gland. The cleavage site of the SP is discussed and compared with the predictive results of the SignalP 3.0 online prediction program.

  8. Surface modification and properties of Bombyx mori silk fibroin films by antimicrobial peptide

    Energy Technology Data Exchange (ETDEWEB)

    Bai Liqiang [Key Laboratory of Advanced Textile Materials and Manufacturing Technology of Ministry of Education, College of Materials and Textile, Zhejiang Sci-Tech University, Xiasha Higher Education Park, Hangzhou 310018 (China); Zhu Liangjun; Min Sijia [College of Animal Sciences, Zhejiang University, Hangzhou 310029 (China); Liu Lin; Cai Yurong [Key Laboratory of Advanced Textile Materials and Manufacturing Technology of Ministry of Education, College of Materials and Textile, Zhejiang Sci-Tech University, Xiasha Higher Education Park, Hangzhou 310018 (China); Yao Juming [Key Laboratory of Advanced Textile Materials and Manufacturing Technology of Ministry of Education, College of Materials and Textile, Zhejiang Sci-Tech University, Xiasha Higher Education Park, Hangzhou 310018 (China)], E-mail: yaoj@zstu.edu.cn

    2008-03-15

    The Bombyx mori silk fibroin films (SFFs) were modified by a Cecropin B (CB) antimicrobial peptide, (NH{sub 2})-NGIVKAGPAIAVLGEAAL-CONH{sub 2}, using the carbodiimide chemistry method. In order to avoid the dissolution of films during the modification procedure, the SFFs were first treated with 60% (v/v) ethanol aqueous solution, resulting a structural transition from unstable silk I to silk II. The investigation of modification conditions showed that the surface-modified SFFs had the satisfied antimicrobial activity and durability when they were activated by EDC.HCl/NHS solution followed by a treatment in CB peptide/PBS buffer (pH 6.5 or 8) solution at ambient temperature for 2 h. Moreover, the surface-modified SFFs showed the smaller contact angle due to the hydrophilic antimicrobial peptides coupled on the film surface, which is essential for the cell adhesion and proliferation. AFM results indicated that the surface roughness of SFFs was considerably increased after the modification by the peptides. The elemental composition analysis results also suggested that the peptides were tightly coupled to the surface of SFFs. This approach may provide a new option to engineer the surface-modified implanted materials preventing the biomaterial-centered infection (BCI)

  9. FLP recombinase-mediated site-specific recombination in silkworm, Bombyx mori.

    Directory of Open Access Journals (Sweden)

    Ding-Pei Long

    Full Text Available A comprehensive understanding of gene function and the production of site-specific genetically modified mutants are two major goals of genetic engineering in the post-genomic era. Although site-specific recombination systems have been powerful tools for genome manipulation of many organisms, they have not yet been established for use in the manipulation of the silkworm Bombyx mori genome. In this study, we achieved site-specific excision of a target gene at predefined chromosomal sites in the silkworm using a FLP/FRT site-specific recombination system. We first constructed two stable transgenic target silkworm strains that both contain a single copy of the transgene construct comprising a target gene expression cassette flanked by FRT sites. Using pre-blastoderm microinjection of a FLP recombinase helper expression vector, 32 G3 site-specific recombinant transgenic individuals were isolated from five of 143 broods. The average frequency of FLP recombinase-mediated site-specific excision in the two target strains genome was approximately 3.5%. This study shows that it is feasible to achieve site-specific recombination in silkworms using the FLP/FRT system. We conclude that the FLP/FRT system is a useful tool for genome manipulation in the silkworm. Furthermore, this is the first reported use of the FLP/FRT system for the genetic manipulation of a lepidopteran genome and thus provides a useful reference for the establishment of genome manipulation technologies in other lepidopteran species.

  10. Immobilization of foreign protein into polyhedra of Bombyx mori nucleopolyhedrovirus (BmNPV)

    Institute of Scientific and Technical Information of China (English)

    Xing-wei XIANG; Rui YANG; Lin CHEN; Xiao-long HU; Shao-fang YU; Cui-ping CAO; Xiao-feng WU

    2012-01-01

    In the late phase of Bombyx mori nucleopolyhedrovirus (BmNPV) infection,a large amount of polyhedra appear in the infected cell nucleolus,these polyhedra being dense protein crystals protecting the incorporated virions from the harsh environment.To investigate whether the foreign protein could be immobilized into the polyhedra of BmNPV,two recombinant baculoviruses were generated by a novel BmNPV polyhedrin-plus (polh+) Bac-toBac system,designated as vBmBac(polh+)-enhanced green fluorescent protein (EGFP) and vBmBac(polh+)-LacZ,which can express the polyhedrin and foreign protein simultaneously.Light microscopy analysis showed that all viruses produced polyhedra of normal appearance.Green fluorescence can be apparently detected on the surface of the vBmBac(polh+)-EGFP polyhedra,but not the BmNPV polyhedra.Fluorescence analysis and anti-desiccation testing confirmed that EGFP was embedded in the polyhedra.As expected,the vBmBac(polh+)-LacZ polyhedre contained an amount of LacZ and had a higher β-galactosidase activity.Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting were also performed to verify if the foreign proteins were immobilized into polyhedra.This study provides a new inspiration for efficient preservation of useful proteins and development of new pesticides with toxic proteins.

  11. Analysis of the activity of virus internal ribosome entry site in silkworm Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    Lupeng Ye; Lanfang Zhuang; Jisheng Li; Zhengying You; Jianshe Liang; Hao Wei; Jianrong Lin

    2013-01-01

    Internal ribosome entry site (IRES) has been widely used in genetic engineering; however,the application in silkworm (Bombyx mori) has hardly been reported.In this study,the biological activity of partial sequence of Encephalomyocardltis virus (EMCV) IRES,Rhopalosiphum padi virus (RhPV)IRES,and the hybrid of IRES of EMCV and RhPV were investigated in Spodoptera frugiperda (Sf9) cell line and silkworm tissues.The hybrid IRES of EMCV and RhPV showed more effective than EMCV IRES or RhPV IRES in promoting downstream gene expression in insect and silkworm.The activities of all IRESs in middle silk gland of silkworm were higher than those in the fat body and posterior silk gland.The hybrid IRES of EMCV and RhPV was integrated into silkworm genome by transgenic technology to test biological activity of IRES.Each of the positive transgenic individuals had significant expression of report gene EGFP.These results suggested that IRES has a potential to be used in the genetic engineering research of silkworm.

  12. Molecular and biochemical characterization of juvenile hormone epoxide hydrolase from the silkworm, Bombyx mori.

    Science.gov (United States)

    Zhang, Qi-Rui; Xu, Wei-Hua; Chen, Fu-Sheng; Li, Sheng

    2005-02-01

    One major route of insect juvenile hormone (JH) degradation is epoxide hydration by JH epoxide hydrolase (JHEH). A full-length cDNA (1536 bp) encoding a microsomal JHEH was isolated from the silkworm, Bombyx mori. Bommo-JHEH cDNA contains an open reading frame encoding a 461-amino acid protein (52 kDa), which reveals a high degree of similarity to the previously reported insect JHEHs. The residues Tyr298, Tyr373, and the HGWP motif corresponding to the oxyanion hole of JHEHs and the residues Asp227, His430, and Glu403 in the catalytic triad are well conserved in Bommo-JHEH. Bommo-JHEH was highly expressed in the fat body, where its mRNA expression pattern was in contrast to the pattern of hemolymph levels of JH during the larval development, suggesting that Bommo-JHEH plays an important role in JH degradation. Recombinant Bommo-JHEH (52 kDa) expressed in Sf9 insect cells was membrane-bound and had a high level of enzyme activity (300-fold over the control activity). This Bommo-JHEH study provides a better understanding of how JH levels are regulated in the domesticated silkworm. PMID:15681225

  13. Reexamination of properties of prophenoloxidase isolated from larval hemolymph of the silkworm Bombyx mori.

    Science.gov (United States)

    Yasuhara, Y; Koizumi, Y; Katagiri, C; Ashida, M

    1995-06-20

    Prophenoloxidase in hemolymph of the silkworm (Bombyx mori) was purified by the method of Ashida (Ashida, M. (1971) Arch. Biochem. Biophys. 144, 749-762) with slight modifications to further increase the purity, and its properties were reinvestigated. The purified prophenoloxidase gave two discrete bands in isoelectric focusing-polyacrylamide gel electrophoresis (IEF-PAGE) (pI 4.95 and 4.98) and in native-polyacrylamide gel electrophoresis with 4.5% separating gel. Each band in IEF-PAGE was separated into two bands in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with mobilities corresponding to 71.5- and 71-kDa polypeptides. In HPLC on octadecyl column the prophenoloxidase preparation gave two well-separated symmetrical peaks (proPO polypeptide I and proPO polypeptide II). The molecular masses of the proPO polypeptides I and II were determined to be 71.5 and 71 kDa in SDS-PAGE and 78,880 and 81,105 Da by matrix-assisted laser desorption ionization mass spectrometry, respectively. Native prophenoloxidase was eluted at a position corresponding to 126-kDa protein in gel permeation chromatography. Amino acid compositions and peptide mappings of proPO polypeptides indicated that both polypeptides differ in their primary structures. These results are discussed in relation to the subunit structure, the presence of bicopper cluster, and the polymorphism of prophenoloxidase in silkworm hemolymph. PMID:7793973

  14. Efficient silkworm expression of human GPCR (nociceptin receptor) by a Bombyx mori bacmid DNA system

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    Kajikawa, Mizuho; Sasaki, Kaori [Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582 (Japan); Wakimoto, Yoshitaro; Toyooka, Masaru [Department of Chemistry and Chemical Biology, Graduate School of Engineering, Gunma University, 1-5-1 Tenjin-cho, Kiryu, Gunma 376-8515 (Japan); Motohashi, Tomoko; Shimojima, Tsukasa [National Institute of Genetics, 1111 Yata, Mishima, Shizuoka 411-8540 (Japan); Takeda, Shigeki [Department of Chemistry and Chemical Biology, Graduate School of Engineering, Gunma University, 1-5-1 Tenjin-cho, Kiryu, Gunma 376-8515 (Japan); Park, Enoch Y. [Laboratory of Biotechnology, Integrated Bioscience Section, Graduate School of Science and Technology, Shizuoka University, 836 Oya, Suruga-ku, Shizuoka, Shizuoka 422-8529 (Japan); Maenaka, Katsumi, E-mail: kmaenaka-umin@umin.net [Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582 (Japan)

    2009-07-31

    Guanine nucleotide-binding protein (G protein) coupled receptors (GPCRs) are frequently expressed by a baculovirus expression vector system (BEVS). We recently established a novel BEVS using the bacmid system of Bombyx mori nucleopolyhedrovirus (BmNPV), which is directly applicable for protein expression in silkworms. Here, we report the first example of GPCR expression in silkworms by the simple injection of BmNPV bacmid DNA. Human nociceptin receptor, an inhibitory GPCR, and its fusion protein with inhibitory G protein alpha subunit (G{sub i}{alpha}) were both successfully expressed in the fat bodies of silkworm larvae as well as in the BmNPV viral fraction. Its yield was much higher than that from Sf9 cells. The microsomal fractions including the nociceptin receptor fusion, which are easily prepared by only centrifugation steps, exhibited [{sup 35}S]GTP{gamma}S-binding activity upon specific stimulation by nociceptin. Therefore, this rapid method is easy-to-use and has a high expression level, and thus will be an important tool for human GPCR production.

  15. Changes in lipid droplet localization during embryogenesis of the silkworm, Bombyx mori.

    Science.gov (United States)

    Yamahama, Yumi; Seno, Keiji; Hariyama, Takahiko

    2008-06-01

    Lipid droplets are considered one of the most important energy sources in lepidopteran eggs during late embryogenesis, but the process of their incorporation into the embryo is as yet unknown. The present study focused on the process of transition of lipid droplets from the extraembryonic yolk to the embryo of the silkworm Bombyx mori, using morphological and biochemical approaches. The morphological study revealed that the incorporation of lipid droplets from the extraembryonic yolk into the embryo occurs at three points and in three different ways during the development of the embryo. Some lipid droplets were translocated directly from the extraembryonic yolk to the embryo before the blastokinesis stage. However, the majority of lipid droplets together with the other components of the extraembryonic yolk were incorporated in the embryo via both morphogenetic inclusion during dorsal closure and ingestion of the extraembyonic yolk by the developing caterpillar prior to hatching. Similar results were obtained from the biochemical study. Thus, we propose that there are three steps in the incorporation of lipid droplets from the extraembryonic yolk into the embryo. In addition, morphological and biochemical data concerning the total amount of lipid droplets in the egg suggested that lipid droplets were mainly consumed during late embryogenesis, seeming to synchronize with tracheal development. PMID:18624568

  16. Existence of prophenoloxidase in wing discs: a source of plasma prophenoloxidase in the silkworm, Bombyx mori.

    Science.gov (United States)

    Diao, Yupu; Lu, Anrui; Yang, Bing; Hu, Wenli; Peng, Qing; Ling, Qing-Zhi; Beerntsen, Brenda T; Söderhäll, Kenneth; Ling, Erjun

    2012-01-01

    In insects, hemocytes are considered as the only source of plasma prophenoloxidase (PPO). PPO also exists in the hemocytes of the hematopoietic organ that is connected to the wing disc of Bombyx mori. It is unknown whether there are other cells or tissues that can produce PPO and release it into the hemolymph besides circulating hemocytes. In this study, we use the silkworm as a model to explore this possibility. Through tissue staining and biochemical assays, we found that wing discs contain PPO that can be released into the culture medium in vitro. An in situ assay showed that some cells in the cavity of wing discs have PPO1 and PPO2 mRNA. We conclude that the hematopoietic organ may wrongly release hemocytes into wing discs since they are connected through many tubes as repost in previous paper. In wing discs, the infiltrating hemocytes produce and release PPO probably through cell lysis and the PPO is later transported into hemolymph. Therefore, this might be another source of plasma PPO in the silkworm: some infiltrated hemocytes sourced from the hematopoietic organ release PPO via wing discs. PMID:22848488

  17. Cloning and characterization of an inhibitor of apoptosis protein (IAP) from Bombyx mori.

    Science.gov (United States)

    Huang, Q; Deveraux, Q L; Maeda, S; Stennicke, H R; Hammock, B D; Reed, J C

    2001-01-15

    We cloned a novel inhibitor of apoptosis protein (IAP) family member, BmIAP, from Bombyx mori BmN cells. BmIAP contains two baculoviral IAP repeat (BIR) domains followed by a RING domain. BmIAP shares striking amino acid sequence similarity with lepidopteran IAPs, SfIAP and TnIAP, and with two baculoviral IAPs, CpIAP and OpIAP, suggesting evolutionary conservation. BmIAP blocks programmed cell death (apoptosis) in Spodoptera frugiperda Sf-21 cells induced by p35 deficient Autographa californica nucleopolyhedrovirus (AcMNPV). This anti-apoptotic function requires both the BIR domains and RING domain of BmIAP. In mammalian cells, BmIAP inhibits Bax induced but not Fas induced apoptosis. Further biochemical data suggest that BmIAP is a specific inhibitor of mammalian caspase-9, an initiator caspase in the mitochondria/cytochrome-c pathway, but not the downstream effector proteases, caspase-3 and caspase-7. These results suggest that suppression of apoptosis by lepidopteran IAPs in insect cells may involve inhibition of an upstream initiator caspase in the conserved mitochondria/cytochrome-c pathway for apoptosis. PMID:11341966

  18. Silkworm (Bombyx mori) hemolymph unable to substitute fetal bovine serum in insect cell culture

    Science.gov (United States)

    Suparto, Irma H.; Khalam, Chandra Nur; Praira, Willy; Sajuthi, Dondin

    2014-03-01

    Fetal Bovine Serum (FBS) in animal cell culture media is an important source of nutrients for cell growth. However, the harvest and collection of FBS cause bioethical concerns. Efforts to reduce and preferably replace FBS with synthetic or other natural alternatives are continually being explored. Hemolymph silkworm (Bombyx mori) contains many nutrients needed for the process of metamorphosis. Therefore, there is possibility as an alternative nutritional supplement for cell culture to reduce the use of FBS. The objective of this study was to evaluate the macrocomponent of hemolymph and the possibility as medium supplement for Spodoptera fugiperda (Sf9) cell culture. Proximate analyses showed that hemolymph contains 89.76% of water, 2.52 mg/mL carbohydrate, 2.35% fat and 55.61 mg/mL protein. Further protein analysis, it consists of 15 fractions containing molecular weight of 22 - 152 kDa. The use of hemolymph as FBS substitution in Sf9 cell culture with various concentrations was unable to maintain and support cell growth. Further research still needed by prior adaptation of the tissue culture to minimal nutrition media before introduction of the hemolymph as supplement.

  19. Differences in nutrient uptake between the fat body and embryonic primary cultures of silkworm (Bombyx mori)

    Institute of Scientific and Technical Information of China (English)

    LEILA MATINDOOST; JALAL J. SENDI; HOORIEH SOLEIMAN JAHI; KAYVAN ETEBARI

    2006-01-01

    Nutrition utilization and by-product formation in cultured insect cells has been investigated in several insect cells and has been of great interest to cell culturists and physiologists. In this research the biochemical changes in embryonic and fat body primary cultures of silkworm, Bombyx mori, have been compared. TC-100 medium supplemented with 10% and 20% FBS was used in embryonic and fat body primary cultures, respectively.Medium was renewed every week and the amount of glucose, uric acid, urea, total protein and alkaline phosphatase were measured in the samples from medium of primary cultures using spectrophotometeric methods. All biochemical macromolecules except uric acid showed significant changes. Glucose decreased in embryonic tissues, while in fat body culture its amount increased. Urea accumulation in embryonic culture was higher than in the fat body cultures. Since urea is a by-product, this accumulation could be due to higher utilization of amino acids. Total protein showed considerable changes and was consumed by embryonic culture more than the fat body' s. Alkaline phosphatase showed stronger activity in embryonic cells.

  20. Comparison of Transformation Efficiency of piggyBac Transposon among Three Different Silkworm Bombyx mori Strains

    Institute of Scientific and Technical Information of China (English)

    Boxiong ZHONG; Jianying LI; Jin'e CHEN; Jian YE; Songdong YU

    2007-01-01

    The transformation rate of three different strains of silkworm Bombyx mori was compared after the introduction of enhanced green fluorescence protein (EGFP)-encoding genes into the silkworm eggs by microinjection of a mixture of piggyBac vector and helper plasmid containing a transposase-encoding sequence. Although there were no significant differences among the three strains in the percentages of fertile moths in microinjected eggs (P=0.1258), the percentages of Go transformed moths in fertile moths and injected eggs were both significantly different (P=0.01368 and P=0.02398, respectively). The transformation rate of the Nistari strain (Indian strain) was significantly higher than that of the other two strains, Golden-yellow-cocoon (Vietnamese strain) and Jiaqiu (Chinese strain), which had similar rate. These results indicate that the transformation efficiency of the piggyBac-based system might vary with silkworm strains with different genetic backgrounds. The presence of endogenous piggyBac-like elements might be an important factor influencing the transformation efficiency of introduced piggyBac-derived vectors, and the diverse amount and activation in different silkworm strains might account for the significant differences.

  1. Melanin pigmentation gives rise to black spots on the wings of the silkworm Bombyx mori.

    Science.gov (United States)

    Ito, Katsuhiko; Yoshikawa, Manabu; Fujii, Takeshi; Tabunoki, Hiroko; Yokoyama, Takeshi

    2016-01-01

    Several mutants of the silkworm Bombyx mori show body color variation at the larval and adult stages. The Wild wing spot (Ws) mutant exhibits a phenotype in which the moth has a spot on the apex of the forewing. In this study, we investigated this trait to elucidate the molecular mechanism underlying the color pattern. Microscopy of the black spot of Ws mutants showed that the pigment emerges in the scales of the wing, and accumulation of the pigment becomes strong just before eclosion. We next examined the relationship between the black spot of the Ws mutant and melanin. The spectrophotometry using alkaline extracts from the black spot in the wing showed the highest absorption intensity at 405nm, which is the absorbance wavelength of melanin. Moreover, inhibition assays for enzymes implicated in melanin synthesis using 3-iodo-l-tyrosine (a tyrosine hydroxylase inhibitor) and L-α-methyl-DOPA (a dopa decarboxylase inhibitor) revealed that treatment with each inhibitor disrupted the pigmentation of the wing of the Ws mutant. On the basis of these results, we analyzed the expression pattern of five genes involved in melanin formation, and found that the expression levels of yellow and laccase2 were increased just before pigmentation, whereas those of DDC, tan, and TH were increased when the apex of the wing turned black. These results showed that melanin pigmentation gives rise to the black spot on the wing. PMID:27405010

  2. Enhancing Effect of Glycerol on the Tensile Properties of Bombyx mori Cocoon Sericin Films

    Directory of Open Access Journals (Sweden)

    Liangjun Zhu

    2011-05-01

    Full Text Available An environmental physical method described herein was developed to improve the tensile properties of Bombyx mori cocoon sericin films, by using the plasticizer of glycerol, which has a nontoxic effect compared with other chemical crosslinkers. The changes in the tensile characteristics and the structure of glycerolated (0–40 wt% of glycerol sericin films were investigated. Sericin films, both in dry and wet states, showed enhanced tensile properties, which might be regulated by the addition of different concentrations of glycerol. The introduction of glycerol results in the higher amorphous structure in sericin films as evidenced by analysis of attenuated total reflection Fourier transform infrared (ATR-FTIR spectra, thermogravimetry (TGA and differential scanning calorimetry (DSC curves. Scanning Electron Microscopy (SEM observation revealed that glycerol was homogeneously blended with sericin molecules when its content was 10 wt%, while a small amount of redundant glycerol emerged on the surface of sericin films when its content was increased to 20 wt% or higher. Our results suggest that the introduction of glycerol is a novel nontoxic strategy which can improve the mechanical features of sericin-based materials and subsequently promote the feasibility of its application in tissue engineering.

  3. Processing and characterization of silk sericin from Bombyx mori and its application in biomaterials and biomedicines.

    Science.gov (United States)

    Cao, Ting-Ting; Zhang, Yu-Qing

    2016-04-01

    Bombyx mori silk is composed of 60-80% fibroin, 15-35% sericin and 1-5% non-sericin component including wax, pigments, sugars and other impurities. For two decades, the protein-based silk fibroin was extensively used in the research and development of medical biomaterials and biomedicines. Sericin is frequently ignored and abandoned as a byproduct or waste in the processing of traditional silk fabrics, silk floss or modern silk biomaterials. However, similar to fibroin, sericin is not only a highly useful biological material, but also a lot of biological activity. Moreover, the non-sericin component present with sericin in the cocoon shell also has a strong biological activity. In this review, the extraction and recovery methods of sericin and the non-sericin component from the cocoon layer are reported, and their composition, properties and biological activity are described to produce a comprehensive report on biomedical materials and biological drugs. In addition, related problems or concerns present in the research and development of sericin are discussed, and a potential application of sericin in sustainable development is also presented. PMID:26838924

  4. Cloning and expression of Bombyx mori silk gland elongation factor 1gamma in Escherichia coli.

    Science.gov (United States)

    Kamiie, Katsuyoshi; Nomura, Yoshitaka; Kobayashi, Satoru; Taira, Hideharu; Kobayashi, Kohmei; Yamashita, Tetsuro; Kidou, Shin-ichiro; Ejiri, Shin-ichiro

    2002-03-01

    Elongation factor 1 (EF-1) from the silk gland of Bombyx mori consists of alpha-, beta-, gamma-, and delta-subunits. EF-1alpha GTP catalyzes the binding of aminoacyl-tRNA to ribosomes concomitant with the hydrolysis of GTP. EF-1betagammadelta catalyzes the exchange of EF-1alpha-bound GDP for exogenous GTP and stimulates the EF-1alpha-dependent binding of aminoacyl-tRNA to ribosomes. EF-1gamma cDNA, which contains an open reading frame (ORF) encoding a polypeptide of 423 amino acid residues, was amplified and cloned by PCR from a silk gland cDNA library. The calculated molecular mass and predicted pI of the product were 48,388 Da and 5.84, respectively. The silk gland EF-1gamma shares 67.3% amino acid identity with Artemia salina EF-lgamma. The N-terminal domain (amino acid residues 1-211) of silk gland EF-lgamma is 29.3% identical to maize glutathione S-transferase. We demonstrated that silk gland EF-lgamma bound to glutathione Sepharose, suggesting that the N-terminal domain of EF-1gamma may have the capacity to bind to glutathione. PMID:12005049

  5. Ectopic expression of ecdysone oxidase impairs tissue degeneration in Bombyx mori

    Science.gov (United States)

    Li, Zhiqian; You, Lang; Zeng, Baosheng; Ling, Lin; Xu, Jun; Chen, Xu; Zhang, Zhongjie; Palli, Subba Reddy; Huang, Yongping; Tan, Anjiang

    2015-01-01

    Metamorphosis in insects includes a series of programmed tissue histolysis and remolding processes that are controlled by two major classes of hormones, juvenile hormones and ecdysteroids. Precise pulses of ecdysteroids (the most active ecdysteroid is 20-hydroxyecdysone, 20E), are regulated by both biosynthesis and metabolism. In this study, we show that ecdysone oxidase (EO), a 20E inactivation enzyme, expresses predominantly in the midgut during the early pupal stage in the lepidopteran model insect, Bombyx mori. Depletion of BmEO using the transgenic CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/RNA-guided Cas9 nucleases) system extended the duration of the final instar larval stage. Ubiquitous transgenic overexpression of BmEO using the Gal4/UAS system induced lethality during the larval–pupal transition. When BmEO was specifically overexpressed in the middle silk gland (MSG), degeneration of MSG at the onset of metamorphosis was blocked. Transmission electron microscope and LysoTracker analyses showed that the autophagy pathway in MSG is inhibited by BmEO ectopic expression. Furthermore, RNA-seq analysis revealed that the genes involved in autophagic cell death and the mTOR signal pathway are affected by overexpression of BmEO. Taken together, BmEO functional studies reported here provide insights into ecdysone regulation of tissue degeneration during metamorphosis. PMID:26041352

  6. Mechanical properties of cocoons constructed consecutively by a single silkworm caterpillar, Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    SHANG XIAO; S.Q.Huang; H.P.Zhao; X.Q.Feng; W.Cui; Z.Lin; M.Q.Xu

    2008-01-01

    Most animals have the ability to adapt, to some extends and in different ways, the variation or disturbance of environment. In our experiments, we forced a silkworm cater-pillar to spin two, three or four thin cocoons by taking it out from the cocoon being constructed. The mechanical prop-erties of these cocoons were studied by static tensile tests and dynamic mechanical thermal analysis. Though exter-nal disturbances may cause the decrease in the total weight of silk spun by the silkworm, a gradual enhancement was interestingly found in the mechanical properties of these thin cocoons. Scanning electron microscopy observations of the fractured specimens of the cocoons showed that there exist several different energy dissipation mechanisms occurred simultaneously at macro-, meso-, and micro-scales, yield-ing a superior capacity of cocoons to adsorb the energy of possible attacks from the outside and to protect efficiently its pupa against damage. Through evolution of millions of years,therefore, the silkworm Bombyx mori seems to have gained the ability to adapt external disturbances and to redesign a new cocoon with optimized protective function when its first cocoon has been damaged for some reasons.

  7. Genome-Wide Identification and Characterization of Carboxypeptidase Genes in Silkworm (Bombyx mori)

    Science.gov (United States)

    Ye, Junhong; Li, Yi; Liu, Hua-Wei; Li, Jifu; Dong, Zhaoming; Xia, Qingyou; Zhao, Ping

    2016-01-01

    The silkworm (Bombyx mori) is an economically-important insect that can secrete silk. Carboxypeptidases have been found in various metazoan species and play important roles in physiological and biochemical reactions. Here, we analyzed the silkworm genome database and characterized 48 carboxypeptidases, including 34 metal carboxypeptidases (BmMCP1–BmMCP34) and 14 serine carboxypeptidases (BmSCP1–BmSCP14), to better understand their diverse functions. Compared to other insects, our results indicated that carboxypeptidases from silkworm have more family members. These silkworm carboxypeptidases could be divided into four families: Peptidase_M2 carboxypeptidases, Peptidase_M14 carboxypeptidases, Peptidase_S10 carboxypeptidases and Peptidase_S28 carboxypeptidases. Microarray analysis showed that the carboxypeptidases had distinct expression patterns, whereas quantitative real-time PCR demonstrated that the expression level of 13 carboxypeptidases significantly decreased after starvation and restored after re-feeding. Overall, our study provides new insights into the functional and evolutionary features of silkworm carboxypeptidases. PMID:27483237

  8. Analysis of differentially expressed genes between fluoride-sensitive and fluoride-endurable individuals in midgut of silkworm, Bombyx mori.

    Science.gov (United States)

    Qian, Heying; Li, Gang; He, Qingling; Zhang, Huaguang; Xu, Anying

    2016-08-15

    Fluoride tolerance is an economically important trait of silkworm. Near-isogenic lines (NILs) of the dominant endurance to fluoride (Def) gene in Bombyx mori has been constructed before. Here, we analyzed the gene expression profiles of midgut of fluoride-sensitive and fluoride-endurable individuals of Def NILs by using high-throughput Illumina sequencing technology and bioinformatics tools, and identified differentially expressed genes between these individuals. A total of 3,612,399 and 3,567,631 clean tags for the libraries of fluoride-endurable and fluoride-sensitive individuals were obtained, which corresponded to 32,933 and 43,976 distinct clean tags, respectively. Analysis of differentially expressed genes indicates that 241 genes are differentially expressed between the two libraries. Among the 241 genes, 30 are up-regulated and 211 are down-regulated in fluoride-endurable individuals. Pathway enrichment analysis demonstrates that genes related to ribosomes, pancreatic secretion, steroid biosynthesis, glutathione metabolism, steroid biosynthesis, and glycerolipid metabolism are down-regulated in fluoride-endurable individuals. qRT-PCR was conducted to confirm the results of the DGE. The present study analyzed differential expression of related genes and tried to find out whether the crucial genes were related to fluoride detoxification which might elucidate fluoride effect and provide a new way in the fluorosis research. PMID:27106117

  9. Localization, expression, and secretion pathway of diapause hormone in embryo and larva of the silkworm, Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    SUN Jiusong; CHEN Fusheng; XU Weihua

    2004-01-01

    In this paper, the distribution and expression of diapause hormone (DH) at mRNA and protein levels in the central nervous system of Bombyx mori embryo and larva were studied using whole-mount in situ hybridization and immunocytochemistry. Whole-mount immunocytochemistry revealed that the distribution of the DH-like immunoreactivity was throughout the central nervous system including the brain, suboesophageal ganglion (SG) and thoracic ganglia (TG); and that the corpus cardiacum and terminal abdominal ganglion may be the site for DH release due to the presence of strong immunoreactivity. In situ hybridization with the probe labeled by digoxigenin shows that the Bom-DH mRNA was also localized in the mandibular, maxillary, labial cell clusters. In addition, a pair of lateral neurons in the SG and a pair of ventral midline neurons in each TG expressing the Bom-DH transcript were also identified. These results were consistent with the localization of Bom-DH mRNA in larva by in situ hybridization and the distribution of the gene by RT-PCR, which is some different from the results reported previously.

  10. Identification, gene expression and immune function of the novel Bm-STAT gene in virus-infected Bombyx mori.

    Science.gov (United States)

    Zhang, Xiaoli; Guo, Rui; Kumar, Dhiraj; Ma, Huanyan; Liu, Jiabin; Hu, Xiaolong; Cao, Guangli; Xue, Renyu; Gong, Chengliang

    2016-02-10

    Genes in the signal transducer and activator of transcription (STAT) family are vital for activities including gene expression and immune response. To investigate the functions of the silkworm Bombyx mori STAT (Bm-STAT) gene in antiviral immunity, two Bm-STAT gene isoforms, Bm-STAT-L for long form and Bm-STAT-S for short form, were cloned. Sequencing showed that the open reading frames were 2313 bp encoding 770 amino acid residues for Bm-STAT-L and 2202 bp encoding 734 amino acid residues for Bm-STAT-S. The C-terminal 42 amino acid residues of Bm-STAT-L were different from the last 7 amino acid residues of Bm-STAT-S. Immunofluorescence showed that Bm-STAT was primarily distributed in the nucleus. Transcription levels of Bm-STAT in different tissues were determined by quantitative PCR, and the results revealed Bm-STAT was mainly expressed in testes. Western blots showed two bands with molecular weights of 70 kDa and 130 kDa in testes, but no bands were detected in ovaries by using anti-Bm-STAT antibody as the primary antibody. Expression of Bm-STAT in hemolymph at 48 h post infection with B. mori macula-like virus (BmMLV) was slightly enhanced compared with controls, suggesting a weak response induced by infection with BmMLV. Hemocyte immunofluorescence showed that Bm-STAT expression was elevated in B. mori nucleopolyhedrovirus (BmNPV)-infected cells. Moreover, resistance of BmN cells to BmNPV was reduced by downregulation of Bm-STAT expression and increased by upregulation. Resistance of BmN cells to BmCPV was not significantly improved by upregulating Bm-STAT expression. Therefore, we concluded that Bm-STAT is a newly identified insect gene of the STAT family. The JAK-STAT pathway has a more specialized role in antiviral defense in silkworms, but JAK-STAT pathway is not triggered in response to all viruses. PMID:26592694

  11. Expression analysis of several antiviral related genes to BmNPV in different resistant strains of silkworm, Bombyx mori.

    Science.gov (United States)

    Cheng, Yang; Wang, Xue-yang; Du, Chang; Gao, Juan; Xu, Jia-ping

    2014-01-01

    Bombyx mori L. (Lepidoptera: Bombycidae) nucleopolyhedrovirus (BmNPV) is a highly pathogenic virus in the sericultural industry, often causing severe damage leading to large economic losses. The immune mechanisms of B. mori against this virus remain obscure. Previous studies had demonstrated Bmlipase-1, BmNox and Bmserine protease-2 showing antiviral activity in vitro, but data on the transcription levels of these proteins in different resistant strains were not reported. In order to determine the resistance level of the four different strains (P50, A35, A40, A53) and gain a better understanding of the mechanism of resistance to BmNPV in B. mori, the relative expression level of the genes coding the three antiviral proteins in larval haemolymph and midgut of different B. mori strains resistant to BmNPV was determined. The results showed that these genes expressed significantly higher in the resistant strains compared to the susceptible strain, and the differential expression levels were consistent with the LC50 values in different strains. The transcription level of the target genes almost all up-regulated in the larvae midgut and down-regulated in the haemolymph. The results indicate the correlation of these genes to BmNPV resistance in B. mori. PMID:25373223

  12. Biochemical characterization and functional analysis of the POU transcription factor POU-M2 of Bombyx mori.

    Science.gov (United States)

    Liu, Lina; Li, Yu; Wang, Yejing; Zhao, Peng; Wei, Shuguang; Li, Zhenzhen; Chang, Huaipu; He, Huawei

    2016-05-01

    POU-M2 is a homeodomain transcription factor which plays important roles in the development and silk synthesis of Bombyx mori. In this study, we expressed, purified and characterized POU-M2 and studied its transcription regulation on fibroin heavy chain gene of Bombyx mori. Gel filtration showed POU-M2 existed as a dimer in solution. Far-UV circular dichroism spectra indicated POU-M2 had a well-defined α-helix structure and the α-helix content was about 26.4%. The thermal unfolding transition of POU-M2 was a cooperative process. Tm, ΔH and ΔS were 45.15±0.2°C, 138.4±0.5KJ/mol and 0.4349±0.04KJ/(mol·K), respectively. Western blotting analysis indicated the expression level of POU-M2 increased slightly from day 3 to day 7 of the fifth instar larvae in the posterior silk gland. POU-M2 was positioned in the nucleus of cells. The luciferase reporter assay demonstrated POU-M2 could stimulate the promoter activity of fibroin heavy chain gene, and the activation effect was dependent on the amount of POU-M2. Our study suggested POU-M2 may be involved in the transcriptional regulation of fibroin heavy chain gene. These findings expand toward a better understanding of the structure of POU-M2 and its function in silk synthesis of Bombyx mori. PMID:26854886

  13. Genome editing of BmFib-H gene provides an empty Bombyx mori silk gland for a highly efficient bioreactor

    OpenAIRE

    Ma, Sanyuan; Shi, Run; Wang, Xiaogang; Liu, Yuanyuan; Chang, Jiasong; Gao, Jie; Lu, Wei; Zhang, Jianduo; ZHAO, PING; Xia, Qingyou

    2014-01-01

    Evolution has produced some remarkable creatures, of which silk gland is a fascinating organ that exists in a variety of insects and almost half of the 34,000 spider species. The impressive ability to secrete huge amount of pure silk protein, and to store proteins at an extremely high concentration (up to 25%) make the silk gland of Bombyx mori hold great promise to be a cost-effective platform for production of recombinant proteins. However, the extremely low production yields of the numerou...

  14. Studies on the sericin 3 of \\kur{Bombyx mori} and cloned sericin into \\kur{Escherichia coli}

    OpenAIRE

    KRŮČEK, Tomáš

    2011-01-01

    The spun-out silk fiber consists of two fibroin filaments that are cemented together by sericin coating. The serine-rich sericins, which make 20-30% of the cocoon silk proteins in Bombyx mori, are dissolved in hot water during silk fiber reeling from the cocoon. The sericin extract is usually discarded. Only small amounts are currently used in cosmetics and lately also as replacement of bovine serum products in the cell culture media. The use in culture media is hindered by poor standardizati...

  15. Participation of D-serine in the development and reproduction of the silkworm Bombyx mori.

    Science.gov (United States)

    Tanigawa, Minoru; Suzuki, Chihiro; Niwano, Kimio; Kanekatsu, Rensuke; Tanaka, Hiroyuki; Horiike, Kihachiro; Hamase, Kenji; Nagata, Yoko

    2016-04-01

    The silkworm Bombyx mori contains high concentrations of free D-serine, an optical isomer of L-serine. To elucidate its function, we first investigated the localization of D-serine in various organs of silkworm larvae, pupae, and adult moths. Using immunohistochemical analysis with an anti-D-serine antibody, we found D-serine in the microvilli of midgut goblet and cylindrical cells and in peripheral matrix components of testicular and ovarian cells. By spectrophotometric analysis, D-serine was also found in the hemolymph and fat body. D-Alanine was not detected in the various organs by immunohistochemistry. Serine racemase, which catalyzes the inter-conversion of L- and D-serine, was found to co-localize with D-serine, and D-serine production from L-serine by intrinsic serine racemase was suggested. O-Phospho-L-serine is an inhibitor of serine racemase, and it was administered to the larvae to reduce the D-serine level. This reagent decreased the midgut caspase-3 level and caused a delay in spermatogenesis and oogenesis. The reagent also decreased mature sperm and egg numbers, suggesting D-serine participation in these processes. D-Serine administration induced an increase in pyruvate levels in testis, midgut, and fat body, indicating conversion of D-serine to pyruvate. On the basis of these results, together with our previous investigation of ATP biosynthesis in testis, we consider the possible involvement of D-serine in ATP synthesis for metamorphosis and reproduction. PMID:26828952

  16. Effect of the sterilization method on the properties of Bombyx mori silk fibroin films

    Energy Technology Data Exchange (ETDEWEB)

    George, Karina A. [Queensland Eye Institute, 41 Annerley Road, South Brisbane, Queensland 4101 (Australia); Institute of Health and Biomedical Innovation, Queensland University of Technology, Kelvin Grove, Queensland 4059 (Australia); Shadforth, Audra M.A. [Queensland Eye Institute, 41 Annerley Road, South Brisbane, Queensland 4101 (Australia); Chirila, Traian V., E-mail: traian.chirila@qei.org.au [Queensland Eye Institute, 41 Annerley Road, South Brisbane, Queensland 4101 (Australia); Australian Institute of Bioengineering and Nanotechnology, University of Queensland, St Lucia, 4072 (Australia); Faculty of Health Sciences, University of Queensland, Herston, Queensland 4006 (Australia); Faculty of Science and Engineering, Queensland University of Technology, Brisbane, Queensland 4001 (Australia); Laurent, Matthieu J. [Queensland Eye Institute, 41 Annerley Road, South Brisbane, Queensland 4101 (Australia); Ecole Superieure d' Ingenieurs de Luminy (ESIL), Universite de la Mediterranee Aix-Marseille II, Luminy case 925 13288, Marseille, Cedex 09 (France); Stephenson, Sally-Anne [Institute of Health and Biomedical Innovation, Queensland University of Technology, Kelvin Grove, Queensland 4059 (Australia); Faculty of Health, Queensland University of Technology, Brisbane, Queensland 4001 (Australia); Edwards, Grant A. [Australian Institute of Bioengineering and Nanotechnology, University of Queensland, St Lucia, 4072 (Australia); Madden, Peter W. [Queensland Eye Institute, 41 Annerley Road, South Brisbane, Queensland 4101 (Australia); Faculty of Health Sciences, University of Queensland, Herston, Queensland 4006 (Australia); and others

    2013-03-01

    We have compared the effects of different sterilization techniques on the properties of Bombyx mori silk fibroin thin films with the view to subsequent use for corneal tissue engineering. The transparency, tensile properties, corneal epithelial cell attachment and degradation of the films were used to evaluate the suitability of certain sterilization techniques including gamma-irradiation (in air or nitrogen), steam treatment and immersion in aqueous ethanol. The investigations showed that gamma-irradiation, performed either in air or in a nitrogen atmosphere, did not significantly alter the properties of films. The films sterilized by gamma-irradiation or by immersion in ethanol had a transparency greater than 98% and tensile properties comparable to human cornea and amniotic membrane, the materials of choice in the reconstruction of ocular surface. Although steam-sterilization produced stronger, stiffer films, they were less transparent, and cell attachment was affected by the variable topography of these films. It was concluded that gamma-irradiation should be considered to be the most suitable method for the sterilization of silk fibroin films, however, the treatment with ethanol is also an acceptable method. - Highlights: Black-Right-Pointing-Pointer The effects of four methods of sterilization on the properties of silk fibroin films were investigated. Black-Right-Pointing-Pointer Steam treatment leads to stiffer films but to lower transparency and variable surface topography. Black-Right-Pointing-Pointer Degradation of fibroin is enhanced in the films that were gamma-irradiated. Black-Right-Pointing-Pointer The effects on mechanical properties are explained through changes in both primary and secondary structure of fibroin. Black-Right-Pointing-Pointer Gamma-irradiation and immersion in aqueous ethanol are suggested as preferred methods of sterilization.

  17. Effect of the sterilization method on the properties of Bombyx mori silk fibroin films

    International Nuclear Information System (INIS)

    We have compared the effects of different sterilization techniques on the properties of Bombyx mori silk fibroin thin films with the view to subsequent use for corneal tissue engineering. The transparency, tensile properties, corneal epithelial cell attachment and degradation of the films were used to evaluate the suitability of certain sterilization techniques including gamma-irradiation (in air or nitrogen), steam treatment and immersion in aqueous ethanol. The investigations showed that gamma-irradiation, performed either in air or in a nitrogen atmosphere, did not significantly alter the properties of films. The films sterilized by gamma-irradiation or by immersion in ethanol had a transparency greater than 98% and tensile properties comparable to human cornea and amniotic membrane, the materials of choice in the reconstruction of ocular surface. Although steam-sterilization produced stronger, stiffer films, they were less transparent, and cell attachment was affected by the variable topography of these films. It was concluded that gamma-irradiation should be considered to be the most suitable method for the sterilization of silk fibroin films, however, the treatment with ethanol is also an acceptable method. - Highlights: ► The effects of four methods of sterilization on the properties of silk fibroin films were investigated. ► Steam treatment leads to stiffer films but to lower transparency and variable surface topography. ► Degradation of fibroin is enhanced in the films that were gamma-irradiated. ► The effects on mechanical properties are explained through changes in both primary and secondary structure of fibroin. ► Gamma-irradiation and immersion in aqueous ethanol are suggested as preferred methods of sterilization.

  18. Recognition of signal peptide by protein translocation machinery in middle silk gland of silkworm Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    Xiuyang Guo; Yi Zhang; Xue Zhang; Shengpeng Wang; Changde Lu

    2008-01-01

    To investigate the functions of signal peptide in protein secretion in the middle silk gland of silkworm Bombyx mori,a series of recombinant Autographa californica multiple nucleopolyhedroviruses containing enhanced green fluorescent protein (egfp) gene,led by sericin-1 promoter and mutated signal peptide coding sequences,were constructed by region-deletions or single amino acid residue deletions.The recombinant Autographa californica multiple nucleopolyhedroviruses were injected into the hemocoele of newly ecdysed fifth-instar silkworm larvae.The expression and secretion of EGFP in the middle silk gland were examined by fluorescence microscopy and Western blot analysis.Results showed that even with a large part (up to 14 amino acid residues) of the ser-1 signal peptide deleted,the expressed EGFP could still be secreted into the cavity of the silk gland.Western blot analysis showed that shortening of the signal peptide from the C-terminal suppressed the maturation of pro-EGFP to EGFP.When 8 amino acid residues were deleted from the C-terminal of the signal peptide (mutant 13 aa),the secretion of EGFP was incomplete,implicating the importance of proper coupling of the h-region and c-region.The deletion of amino acid residue(s) in the h-region did not affect the secretion of EGFP,indicating that the recognition of signal peptide by translocation machinery was mainly by a structural domain,but not by special amino acid residue(s).Furthermore,the deletion of Arg2 or replacement with Asp in the n-region of the signal peptide did not influence secretion of EGFP,suggesting that a positive charge is not crucial.

  19. Monosaccharide profiling of silkworm (Bombyx mori L.) nervous system during development and aging.

    Science.gov (United States)

    Soya, Seçkin; Şahar, Umut; Karaçalı, Sabire

    2016-09-01

    Glycoconjugates have various functions in differentiation, development, aging and in all aspects of normal functioning of organisms. The reason for increased research on this topic is that glycoconjugates locate mostly on the cell surface and play crucial biological roles in the nervous system including brain development, synaptic plasticity, learning, and memory. Considering their roles in the nervous system, information about their existence in the insect nervous system is rather sparse. Therefore, in order to detect monosaccharide content of N- and O-glycans, we carried out capLC-ESI-MS/MS analysis to determine the concentration changes of glucose, mannose, galactose, N-acetylglucosamine (GlcNAc), N-acetylgalactosamine (GalNAc), fucose, xylose, arabinose, and ribose monosaccharides in the nervous system of Bombyx mori during development and aging processes. In addition to LC-MS, lectin blotting was done to detect quantitative changes in N- and O-glycans. Developmental stages were selected as 3rd (the youngest sample), 5th (young) larval instar, motionless prepupa (the oldest sample), and pupa (adult development). Derivatization of monosaccharides was performed with a solution of PMP agent and analyzed with capLC-ESI-MS/MS. For lectin blotting, determination of glycan types was carried out with Galanthus nivalis agglutinin and Peanut agglutinin lectins. In all stages, the most abundant monosaccharide was glucose. Although all monosaccharides were present most abundantly in the youngest stage (3rd instar), they are generally reduced gradually during the aging process. It was observed that amounts of monosaccharides increased again in the pupa stage. According to lectin blotting, N- and O-linked glycoproteins expressions were different and there were some specific glycoprotein expression differences between stages. These findings suggest that the glycosylation state of proteins in the nervous system changes during development and aging in insects in a similar

  20. Disappearance of chorion proteins from Bombyx mori eggs treated with HCl solution to prevent diapause.

    Science.gov (United States)

    Tsurumaru, Shuichiro; Kawamori, Akihito; Mitsumasu, Kanako; Niimi, Teruyuki; Imai, Kunio; Yamashita, Okitsugu; Yaginuma, Toshinobu

    2010-12-01

    Bombyx mori eggs enter diapause immediately after completion of mesoderm segregation. HCl treatment of approximately 24-hour-old eggs (germband formation stage) is well known to be the most effective procedure to prevent entry into diapause, although the molecular mechanism remains unclear. In this study, we examined the protein composition of diapausing and nondiapausing eggs after various HCl treatments known to prevent or break diapause and found that proteins of approximately 11 and 8 kDa disappeared immediately after HCl treatment. Partial amino acid sequences of these proteins indicated that they were members of the chorion class A protein L12 family synthesized in follicle cells. Under the hypothesis that the chorion provides a barrier to oxygen, dechorionation of diapausing eggs induces resumption of embryonic development. Hence, to test this and other hypotheses about the function of these proteins, we used 20% SDS-PAGE with Coomassie Brilliant Blue staining to trace their disappearance from embryos and eggshells after treatment with HCl under different conditions and on polyvoltine, univoltine, and bivoltine silkworm races. Even when 10-day-old diapausing eggs were treated with HCl, which did not break diapause, the 11 and 8 kDa proteins disappeared. Our results suggest that disappearance of these proteins is not directly associated with preventing entry into or breaking a diapause state. Nevertheless, our results cannot completely rule out the possibility that the 11 and 8 kDa proteins function to block permeability of O(2) during the period when HCl treatment is physiologically effective to prevent diapause so that after the diapause system is established within the egg, even removing the 11 and 8 kDa proteins may not affect to prevent diapause. We also discuss the role of these proteins in choriogenesis. PMID:20637776

  1. Ca2+-induced self-assembly of Bombyx mori silk sericin into a nanofibrous network-like protein matrix for directing controlled nucleation of hydroxylapatite nano-needles

    OpenAIRE

    Yang, Mingying; Zhou, Guanshan; Shuai, Yajun; Wang, Jie; Zhu, Liangjun; Mao, Chuanbin

    2015-01-01

    Bone biomineralization is a well-regulated protein-mediated process where hydroxylapatite (HAP) crystals are nucleated with preferred orientation within self-assembled protein matrix. Mimicking this process is a promising approach to the production of bone-like protein/mineral nanocomposites for bone repair and regeneration. Towards the goal of fabricating such nanocomposites from sericin, a protein spun by Bombyx mori (B.mori) silkworm, and bone mineral HAP, for the first time we investigate...

  2. Proteome profiling reveals tissue-specific protein expression in male and female accessory glands of the silkworm, Bombyx mori.

    Science.gov (United States)

    Dong, Zhaoming; Wang, Xiaohuan; Zhang, Yan; Zhang, Liping; Chen, Quanmei; Zhang, Xiaolu; Zhao, Ping; Xia, Qingyou

    2016-05-01

    Male accessory gland (MAG) and female accessory gland (FAG) of the reproductive system are, respectively, responsible for producing seminal proteins and adhesive proteins during copulation and ovulation. Seminal proteins are ejaculated to female along with sperms, whereas adhesive proteins are excreted along with eggs. Proteins from the male and female reproductive organs are usually indicative of rapid adaptive evolution. Understanding the reproductive isolation and species divergence requires identifying reproduction-related proteins from many different species. Here, we present our proteomic analyses of male and female accessory glands of the silkworm, Bombyx mori. Using LC/MS-MS, we identified 2133 MAG proteins and 1872 FAG proteins. In total, 652 proteins were significant more abundant in the MAG than in the FAG, including growth factors, odorant-binding proteins, enzymes, and proteins of unknown function. Growth factors and odorant-binding proteins are potential signaling molecules, whereas most of proteins of unknown function were found to be Lepidoptera-specific proteins with high evolutionary rates. Microarray experiments and semi-quantitative RT-PCR validated that MAG-specific proteins were expressed exclusively in male moths. Totally, 192 proteins were considered as FAG-specific proteins, including protease inhibitors, enzymes, and other proteins. Protease inhibitors were found to be the most abundant FAG-specific proteins, which may protect eggs from infection by inhibiting pathogen-derived proteases. These results provide comprehensive insights into copulation and oviposition. Moreover, the newly identified Lepidoptera-specific MAG proteins provide useful data for future research on the evolution of reproductive proteins in insects. PMID:26822097

  3. Screening for the genes involved in bombykol biosynthesis: Identification and functional characterization of Bombyx mori acyl carrier protein (BmACP

    Directory of Open Access Journals (Sweden)

    ShogoMatsumoto

    2011-12-01

    Full Text Available Species-specific sex pheromones released by female moths to attract conspecific male moths are synthesized de novo in the pheromone gland (PG via fatty acid synthesis (FAS. Biosynthesis of moth sex pheromones is usually regulated by a neurohormone termed pheromone biosynthesis activating neuropeptide (PBAN, a 33-aa peptide that originates in the subesophageal ganglion. In the silkmoth, Bombyx mori, cytoplasmic lipid droplets (LDs, which store the sex pheromone (bombykol precursor fatty acid, accumulate in PG cells prior to eclosion. PBAN activation of the PBAN receptor stimulates lipolysis of the stored LD triacylglycerols (TAGs resulting in release of the bombykol precursor for final modification. While we have previously characterized a number of molecules involved in bombykol biosynthesis, little is known about the mechanisms of PBAN signaling that regulate the TAG lipolysis in PG cells. In the current study, we sought to further identify genes involved in bombykol biosynthesis as well as PBAN signaling, by using a subset of 312 expressed sequence tag (EST clones that are in either our B. mori PG cDNA library or the public B. mori EST databases, SilkBase and CYBERGATE, and which are preferentially expressed in the PG. Using RT-PCR expression analysis and an RNAi screening approach, we have identified another 8 EST clones involved in bombykol biosynthesis. Furthermore, we have determined the functional role of a clone designated BmACP that encodes B. mori acyl carrier protein (ACP. Our results indicate that BmACP plays an essential role in the biosynthesis of the bombykol precursor fatty acid via the canonical FAS pathway during pheromonogenesis.

  4. Identification of lipases involved in PBAN stimulated pheromone production in Bombyx mori using the DGE and RNAi approaches.

    Directory of Open Access Journals (Sweden)

    Mengfang Du

    Full Text Available BACKGROUND: Pheromone biosynthesis activating neuropeptide (PBAN is a neurohormone that regulates sex pheromone synthesis in female moths. Bombyx mori is a model organism that has been used to explore the signal transduction pattern of PBAN, which is mediated by a G-protein coupled receptor (GPCR. Although significant progress has been made in elucidating PBAN-regulated lipolysis that releases the precursor of the sex pheromone, little is known about the molecular components involved in this step. To better elucidate the molecular mechanisms of PBAN-stimulated lipolysis of cytoplasmic lipid droplets (LDs, the associated lipase genes involved in PBAN- regulated sex pheromone biosynthesis were identified using digital gene expression (DGE and subsequent RNA interference (RNAi. RESULTS: Three DGE libraries were constructed from pheromone glands (PGs at different developed stages, namely, 72 hours before eclosion (-72 h, new emergence (0 h and 72 h after eclosion (72 h, to investigate the gene expression profiles during PG development. The DGE evaluated over 5.6 million clean tags in each PG sample and revealed numerous genes that were differentially expressed at these stages. Most importantly, seven lipases were found to be richly expressed during the key stage of sex pheromone synthesis and release (new emergence. RNAi-mediated knockdown confirmed for the first time that four of these seven lipases play important roles in sex pheromone synthesis. CONCLUSION: This study has identified four lipases directly involved in PBAN-stimulated sex pheromone biosynthesis, which improve our understanding of the lipases involved in releasing bombykol precursors from triacylglycerols (TAGs within the cytoplasmic LDs.

  5. Stimulation of orphan nuclear receptor HR38 gene expression by PTTH in prothoracic glands of the silkworm, Bombyx mori.

    Science.gov (United States)

    Gu, Shi-Hong; Hsieh, Yun-Chih; Lin, Pei-Ling

    2016-07-01

    A complex signaling network appears to be involved in prothoracicotropic hormone (PTTH)-stimulated ecdysteroidogenesis in insect prothoracic glands (PGs). Less is known about the genomic action of PTTH signaling. In the present study, we investigated the effect of PTTH on the expression of Bombyx mori HR38, an immediate early gene (IEG) identified in insect systems. Our results showed that treatment of B. mori PGs with PTTH in vitro resulted in a rapid increase in HR38 expression. Injection of PTTH into day-5 last instar larvae also greatly increased HR38 expression, verifying the in vitro effect. Cycloheximide did not affect induction of HR38 expression, suggesting that protein synthesis is not required for PTTH's effect. A mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) kinase (MEK) inhibitor (U0126), and a phosphoinositide 3-kinase (PI3K) inhibitor (LY294002), partially inhibited PTTH-stimulated HR38 expression, implying the involvement of both the ERK and PI3K signaling pathways. When PGs were treated with agents that directly elevate the intracellular Ca(2+) concentration (either A23187 or thapsigargin), an increase in HR38 expression was also detected, indicating that Ca(2+) is involved in PTTH-stimulated HR38 gene expression. A Western blot analysis showed that PTTH treatment increased the HR38 protein level, and protein levels showed a dramatic increase during the later stages of the last larval instar. Expression of HR38 transcription in response to PTTH appeared to undergo development-specific changes. Treatment with ecdysone in vitro did not affect HR38 expression. However, 20-hydroxyecdysone treatment decreased HR38 expression. Taken together, these results demonstrate that HR38 is a PTTH-stimulated IEG that is, at least in part, induced through Ca(2+)/ERK and PI3K signaling. The present study proposes a potential cross talk mechanism between PTTH and ecdysone signaling to regulate insect development and lays a

  6. Molecular and enzymatic characterization of two enzymes BmPCD and BmDHPR involving in the regeneration pathway of tetrahydrobiopterin from the silkworm Bombyx mori.

    Science.gov (United States)

    Li, Wentian; Gong, Meixia; Shu, Rui; Li, Xin; Gao, Junshan; Meng, Yan

    2015-08-01

    Tetrahydrobiopterin (BH4) is an essential cofactor of aromatic amino acid hydroxylases and nitric oxide synthase so that BH4 plays a key role in many biological processes. BH4 deficiency is associated with numerous metabolic syndromes and neuropsychological disorders. BH4 concentration in mammals is maintained through a de novo synthesis pathway and a regeneration pathway. Previous studies showed that the de novo pathway of BH4 is similar between insects and mammals. However, knowledge about the regeneration pathway of BH4 (RPB) is very limited in insects. Several mutants in the silkworm Bombyx mori have been approved to be associated with BH4 deficiency, which are good models to research on the RPB in insects. In this study, homologous genes encoding two enzymes, pterin-4a-carbinolamine dehydratase (PCD) and dihydropteridine reductase (DHPR) involving in RPB have been cloned and identified from B. mori. Enzymatic activity of DHPR was found in the fat body of wild type silkworm larvae. Together with the transcription profiles, it was indicated that BmPcd and BmDhpr might normally act in the RPB of B. mori and the expression of BmDhpr was activated in the brain and sexual glands while BmPcd was expressed in a wider special pattern when the de novo pathway of BH4 was lacked in lemon. Biochemical analyses showed that the recombinant BmDHPR exhibited high enzymatic activity and more suitable parameters to the coenzyme of NADH in vitro. The results in this report give new information about the RPB in B. mori and help in better understanding insect BH4 biosynthetic networks. PMID:25899859

  7. New insights into the catalytic mechanism of Bombyx mori prostaglandin E synthase gained from structure–function analysis

    Energy Technology Data Exchange (ETDEWEB)

    Yamamoto, Kohji, E-mail: yamamok@agr.kyushu-u.ac.jp [Faculty of Agriculture, Kyushu University Graduate School, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581 (Japan); Suzuki, Mamoru; Higashiura, Akifumi [Institute for Protein Research, Osaka University, Suita 565-0871 (Japan); Aritake, Kosuke; Urade, Yoshihiro; Uodome, Nobuko [Department of Molecular Behavioral Biology, Osaka Bioscience Institute, 6-2-4 Furuedai, Suita, Osaka 565-0874 (Japan); Hossain, MD. Tofazzal [Faculty of Agriculture, Kyushu University Graduate School, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581 (Japan); Nakagawa, Atsushi [Institute for Protein Research, Osaka University, Suita 565-0871 (Japan)

    2013-11-01

    Highlights: •Structure of Bombyx mori prostaglandin E synthase is determined. •Bound glutathione sulfonic acid is located at the glutathione-binding site. •Electron-sharing network is present in this protein. •This network includes Asn95, Asp96, and Arg98. •Site-directed mutagenesis reveals that the residues contribute to the catalytic activity. -- Abstract: Prostaglandin E synthase (PGES) catalyzes the isomerization of PGH{sub 2} to PGE{sub 2}. We previously reported the identification and structural characterization of Bombyx mori PGES (bmPGES), which belongs to Sigma-class glutathione transferase. Here, we extend these studies by determining the structure of bmPGES in complex with glutathione sulfonic acid (GTS) at a resolution of 1.37 Å using X-ray crystallography. GTS localized to the glutathione-binding site. We found that electron-sharing network of bmPGES includes Asn95, Asp96, and Arg98. Site-directed mutagenesis of these residues to create mutant forms of bmPGES mutants indicate that they contribute to catalytic activity. These results are, to our knowledge, the first to reveal the presence of an electron-sharing network in bmPGES.

  8. Some autophagic and apoptotic features of programmed cell death in the anterior silk glands of the silkworm, Bombyx mori.

    Science.gov (United States)

    Goncu, Ebru; Parlak, Osman

    2008-11-01

    Programmed cell death has been subdivided into two major groups: apoptosis and autophagic cell death. The anterior silk gland of Bombyx mori degenerates during larval-pupal metamorphosis. Our findings indicate that two types of programmed cell death features are observed during this physiological process. During the prepupal period, pyknosis of the nucleus, cell detachment,and membrane blebbing occur and they are the first signs of programmed cell death in the anterior silk glands. According to previous studies, all of these morphological appearances are common for both cell-death types. Autophagy features are also exhibited during the prepupal period. Levels of one of the lysosomal marker enzymes-acid phosphatase-are high during this period then decrease gradually. Vacuole formation begins to appear first at the basal surface of the cell, then expands to the apical surface just before the larval pupal ecdysis. After larval-pupal ecdysis, DNA fragmentation, which is the obvious biochemical marker of apoptosis, is detected in agarose gel electrophoresis, which also shows that caspase-like enzyme activities occur during the programmed cell death process of the anterior silk glands. Apoptosis and autophagic cell death interact with each other during the degeneration process of the anterior silk gland in Bombyx mori and this interaction occurs at a late phase of cell death. We suggest that apoptotic cell death only is not enough for whole gland degeneration and that more effective degeneration occurs with this cooperation. PMID:18838861

  9. New insights into the catalytic mechanism of Bombyx mori prostaglandin E synthase gained from structure–function analysis

    International Nuclear Information System (INIS)

    Highlights: •Structure of Bombyx mori prostaglandin E synthase is determined. •Bound glutathione sulfonic acid is located at the glutathione-binding site. •Electron-sharing network is present in this protein. •This network includes Asn95, Asp96, and Arg98. •Site-directed mutagenesis reveals that the residues contribute to the catalytic activity. -- Abstract: Prostaglandin E synthase (PGES) catalyzes the isomerization of PGH2 to PGE2. We previously reported the identification and structural characterization of Bombyx mori PGES (bmPGES), which belongs to Sigma-class glutathione transferase. Here, we extend these studies by determining the structure of bmPGES in complex with glutathione sulfonic acid (GTS) at a resolution of 1.37 Å using X-ray crystallography. GTS localized to the glutathione-binding site. We found that electron-sharing network of bmPGES includes Asn95, Asp96, and Arg98. Site-directed mutagenesis of these residues to create mutant forms of bmPGES mutants indicate that they contribute to catalytic activity. These results are, to our knowledge, the first to reveal the presence of an electron-sharing network in bmPGES

  10. Expression of functional human (pro)renin receptor in silkworm (Bombyx mori) larvae using BmMNPV bacmid.

    Science.gov (United States)

    Du, Dongning; Kato, Tatsuya; Nabi, A H M Nurun; Suzuki, Fumiaki; Park, Enoch Y

    2008-03-01

    The circulating RA (renin-angiotensin) system is essential for the regulation of blood pressure and electrolyte balance. Recently, plasma prorenin has been reported to significantly increase its level in diabetes and to be possibly non-proteolytically activated by binding to the PRR [(pro)renin receptor] on the cell membrane reported in several tissues during circulation. Although many pathological aspects have been researched, there is a lack of sufficient information on the biochemical structure and biological function of this hPRR (human PRR) because of the difficulty in increasing hPRR expression. In the present study, GFP(uv)-hPRR (hPRR fused with green fluorescence protein when excited with long-wave UV light) was successfully expressed by using BmMNPV (Bombyx mori multiple nucleopolyhedrovirus) bacmid DNA in silkworm (Bombyx mori) larvae. Some of the hPRR was expressed in the haemolymph of silkworm larvae and some of the hPRR was located in the fat body of silkworm larvae. The binding ability of hPRR expressed in the haemolymph and fat body with renin or prorenin was analysed by ELISA and surface plasmon resonance using a biosensor respectively. These binding assays suggest that the expressed hPRR has a functional bioactivity. hPRR preparation in silkworm larvae would, therefore, be useful for biochemical and biomedical researches related to PRR. PMID:17705788

  11. Regulation of Silk Genes by Hox and Homeodomain Proteins in the Terminal Differentiated Silk Gland of the Silkworm Bombyx mori

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    Shigeharu Takiya

    2016-05-01

    Full Text Available The silk gland of the silkworm Bombyx mori is a long tubular organ that is divided into several subparts along its anteroposterior (AP axis. As a trait of terminal differentiation of the silk gland, several silk protein genes are expressed with unique regional specificities. Most of the Hox and some of the homeobox genes are also expressed in the differentiated silk gland with regional specificities. The expression patterns of Hox genes in the silk gland roughly correspond to those in embryogenesis showing “colinearity”. The central Hox class protein Antennapedia (Antp directly regulates the expression of several middle silk gland–specific silk genes, whereas the Lin-1/Isl-1/Mec3 (LIM-homeodomain transcriptional factor Arrowhead (Awh regulates the expression of posterior silk gland–specific genes for silk fiber proteins. We summarize our results and discuss the usefulness of the silk gland of Bombyx mori for analyzing the function of Hox genes. Further analyses of the regulatory mechanisms underlying the region-specific expression of silk genes will provide novel insights into the molecular bases for target-gene selection and regulation by Hox and homeodomain proteins.

  12. Transcriptome analysis of integument differentially expressed genes in the pigment mutant (quail during molting of silkworm, Bombyx mori.

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    Hongyi Nie

    Full Text Available In the silkworm Bombyx mori, pigment mutants with diverse body colors have been maintained throughout domestication for about 5000 years. The silkworm larval body color is formed through the mutual interaction of melanin, ommochromes, pteridines and uric acid. These pigments/compounds are synthesized by the cooperative action of various genes and enzymes. Previous reports showed that melanin, ommochrome and pteridine are increased in silkworm quail (q mutants. To understand the pigment increase and alterations in pigment synthesis in q mutant, transcriptome profiles of the silkworm integument were investigated at 16 h after head capsule slippage in the fourth molt in q mutants and wild-type (Dazao. Compared to the wild-type, 1161 genes were differentially expressed in the q mutant. Of these modulated genes, 62.4% (725 genes were upregulated and 37.6% (436 genes were downregulated in the q mutant. The molecular function of differently expressed genes was analyzed by Blast2GO. The results showed that upregulated genes were mainly involved in protein binding, small molecule binding, transferase activity, nucleic acid binding, specific DNA-binding transcription factor activity and chromatin binding, while exclusively down-expressed genes functioned in oxidoreductase activity, cofactor binding, tetrapyrrole binding, peroxidase activity and pigment binding. We focused on genes related to melanin, pteridine and ommochrome biosynthesis; transport of uric acid; and juvenile hormone metabolism because of their importance in integument coloration during molting. This study identified differently expressed genes implicated in silkworm integument formation and pigmentation using silkworm q mutant. The results estimated the number and types of genes that drive new integument formation.

  13. A genome-wide survey for host response of silkworm, Bombyx mori during pathogen Bacillus bombyseptieus infection.

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    Lulin Huang

    Full Text Available Host-pathogen interactions are complex relationships, and a central challenge is to reveal the interactions between pathogens and their hosts. Bacillus bombysepticus (Bb which can produces spores and parasporal crystals was firstly separated from the corpses of the infected silkworms (Bombyx mori. Bb naturally infects the silkworm can cause an acute fuliginosa septicaemia and kill the silkworm larvae generally within one day in the hot and humid season. Bb pathogen of the silkworm can be used for investigating the host responses after the infection. Gene expression profiling during four time-points of silkworm whole larvae after Bb infection was performed to gain insight into the mechanism of Bb-associated host whole body effect. Genome-wide survey of the host genes demonstrated many genes and pathways modulated after the infection. GO analysis of the induced genes indicated that their functions could be divided into 14 categories. KEGG pathway analysis identified that six types of basal metabolic pathway were regulated, including genetic information processing and transcription, carbohydrate metabolism, amino acid and nitrogen metabolism, nucleotide metabolism, metabolism of cofactors and vitamins, and xenobiotic biodegradation and metabolism. Similar to Bacillus thuringiensis (Bt, Bb can also induce a silkworm poisoning-related response. In this process, genes encoding midgut peritrophic membrane proteins, aminopeptidase N receptors and sodium/calcium exchange protein showed modulation. For the first time, we found that Bb induced a lot of genes involved in juvenile hormone synthesis and metabolism pathway upregulated. Bb also triggered the host immune responses, including cellular immune response and serine protease cascade melanization response. Real time PCR analysis showed that Bb can induce the silkworm systemic immune response, mainly by the Toll pathway. Anti-microorganism peptides (AMPs, including of Attacin, Lebocin, Enbocin, Gloverin

  14. Location of the Bombyx mori aminopeptidase N type 1 binding site on Bacillus thuringiensis Cry1Aa toxin.

    Science.gov (United States)

    Atsumi, Shogo; Mizuno, Eri; Hara, Hirotaka; Nakanishi, Kazuko; Kitami, Madoka; Miura, Nami; Tabunoki, Hiroko; Watanabe, Ayako; Sato, Ryoichi

    2005-07-01

    We analyzed the binding site on Cry1Aa toxin for the Cry1Aa receptor in Bombyx mori, 115-kDa aminopeptidase N type 1 (BmAPN1) (K. Nakanishi, K. Yaoi, Y. Nagino, H. Hara, M. Kitami, S. Atsumi, N. Miura, and R. Sato, FEBS Lett. 519:215-220, 2002), by using monoclonal antibodies (MAbs) that block binding between the binding site and the receptor. First, we produced a series of MAbs against Cry1Aa and obtained two MAbs, MAbs 2C2 and 1B10, that were capable of blocking the binding between Cry1Aa and BmAPN1 (blocking MAbs). The epitope of the Fab fragments of MAb 2C2 overlapped the BmAPN1 binding site, whereas the epitope of the Fab fragments of MAb 1B10 did not overlap but was located close to the binding site. Using three approaches for epitope mapping, we identified two candidate epitopes for the blocking MAbs on Cry1Aa. We constructed two Cry1Aa toxin mutants by substituting a cysteine on the toxin surface at each of the two candidate epitopes, and the small blocking molecule N-(9-acridinyl)maleimide (NAM) was introduced at each cysteine substitution to determine the true epitope. The Cry1Aa mutant with NAM bound to Cys582 did not bind either of the two blocking MAbs, suggesting that the true epitope for each of the blocking MAbs was located at the site containing Val582, which also consisted of 508STLRVN513 and 582VFTLSAHV589. These results indicated that the BmAPN1 binding site overlapped part of the region blocked by MAb 2C2 that was close to but excluded the actual epitope of MAb 2C2 on domain III of Cry1Aa toxin. We also discuss another area on Cry1Aa toxin as a new candidate site for BmAPN1 binding. PMID:16000811

  15. Genome Sequence of a Novel Iflavirus from mRNA Sequencing of the Pupa of Bombyx mori Inoculated with Cordyceps militaris.

    Science.gov (United States)

    Suzuki, Tomohiro; Takeshima, Yoshino; Mikamoto, Toshiyuki; Saeki, Jun-David; Kato, Tatsuya; Park, Enoch Y; Kawagishi, Hirokazu; Dohra, Hideo

    2015-01-01

    We discovered a novel iflavirus from the transcriptome of the Bombyx mori pupa inoculated with the insect-pathogenic fungus Cordyceps militaris. The assembled iflavirus genome has 10,119 nucleotides, with a 3'-polyadenylated tail, and it encodes a polyprotein composed of 3,004 amino acids. PMID:26383664

  16. Crystal structure of Bombyx mori arylphorins reveals a 3:3 heterohexamer with multiple papain cleavage sites.

    Science.gov (United States)

    Hou, Yong; Li, Jianwei; Li, Yi; Dong, Zhaoming; Xia, Qingyou; Yuan, Y Adam

    2014-06-01

    In holometabolous insects, the accumulation and utilization of storage proteins (SPs), including arylphorins and methionine-rich proteins, are critical for the insect metamorphosis. SPs function as amino acids reserves, which are synthesized in fat body, secreted into the larval hemolymph and taken up by fat body shortly before pupation. However, the detailed molecular mechanisms of digestion and utilization of SPs during development are largely unknown. Here, we report the crystal structure of Bombyx mori arylphorins at 2.8 Å, which displays a heterohexameric structural arrangement formed by trimerization of dimers comprising two structural similar arylphorins. Our limited proteolysis assay and microarray data strongly suggest that papain-like proteases are the major players for B. mori arylphorins digestion in vitro and in vivo. Consistent with the biochemical data, dozens of papain cleavage sites are mapped on the surface of the heterohexameric structure of B. mori arylphorins. Hence, our results provide the insightful information to understand the metamorphosis of holometabolous insects at molecular level. PMID:24639361

  17. Genome editing of BmFib-H gene provides an empty Bombyx mori silk gland for a highly efficient bioreactor.

    Science.gov (United States)

    Ma, Sanyuan; Shi, Run; Wang, Xiaogang; Liu, Yuanyuan; Chang, Jiasong; Gao, Jie; Lu, Wei; Zhang, Jianduo; Zhao, Ping; Xia, Qingyou

    2014-01-01

    Evolution has produced some remarkable creatures, of which silk gland is a fascinating organ that exists in a variety of insects and almost half of the 34,000 spider species. The impressive ability to secrete huge amount of pure silk protein, and to store proteins at an extremely high concentration (up to 25%) make the silk gland of Bombyx mori hold great promise to be a cost-effective platform for production of recombinant proteins. However, the extremely low production yields of the numerous reported expression systems greatly hindered the exploration and application of silk gland bioreactors. Using customized zinc finger nucleases (ZFN), we successfully performed genome editing of Bmfib-H gene, which encodes the largest and most abundant silk protein, in B. mori with efficiency higher than any previously reported. The resulted Bmfib-H knocked-out B. mori showed a smaller and empty silk gland, abnormally developed posterior silk gland cells, an extremely thin cocoon that contain only sericin proteins, and a slightly heavier pupae. We also showed that removal of endogenous Bmfib-H protein could significantly increase the expression level of exogenous protein. Furthermore, we demonstrated that the bioreactor is suitable for large scale production of protein-based materials. PMID:25359576

  18. Genome-Wide Analysis of Host Responses to Four Different Types of Microorganisms in Bombyx Mori (Lepidoptera: Bombycidae).

    Science.gov (United States)

    Cheng, Tingcai; Lin, Ping; Huang, Lulin; Wu, Yuqian; Jin, Shengkai; Liu, Chun; Xia, Qingyou

    2016-01-01

    Several pathogenic microorganisms have been used to investigate the genome-wide transcriptional responses of Bombyx mori to infection. However, studies have so far each focused on one microorganism, and systematic genome-wide comparison of transcriptional responses to different pathogenic microorganisms has not been undertaken. Here, we surveyed transcriptional responses of B. mori to its natural bacterial, viral, and fungal pathogens, Bacillus bombyseptieus, B. mori nucleopolyhedrovirus (BmNPV), and Beauveria bassiana, respectively, and to nonpathogenic Escherichia coli, by microarray analysis. In total, the expression of 2,436, 1,804, 1,743, and 912 B. mori genes was modulated by infection with B. bombyseptieus, BmNPV, B. bassiana, and E. coli, respectively. Notably, the expression of 620, 400, 177, or 165 of these genes was only modulated by infection with B. bombyseptieus, BmNPV, B. bassiana, or E. coli, respectively. In contrast to the expression of genes related to juvenile hormone synthesis and metabolism, that of genes encoding juvenile hormone binding proteins was microorganism-specific. Three basal metabolic pathways were modulated by infection with any of the four microorganisms, and 3, 14, 5, and 2 metabolic pathways were specifically modulated by infection with B. bombyseptieus, BmNPV, B. bassiana, and E. coli, respectively. Interestingly, BmNPV infection modulated the JAK/STAT signaling pathway, whereas both the Imd and Toll signaling pathways were modulated by infection with B. bombyseptieus, B. bassiana, or E. coli These results elucidate potential molecular mechanisms of the host response to different microorganisms, and provide a foundation for further work on host-pathogen interaction. PMID:27382132

  19. The wings of Bombyx mori develop from larval discs exhibiting an early differentiated state: a preliminary report

    Indian Academy of Sciences (India)

    Madhuri Kango-Singh; Amit Singh; K P Gopinathan

    2001-06-01

    Lepidopteran insects present a complex organization of appendages which develop by various mechanisms. In the mulberry silkworm, Bombyx mori a pair of meso- and meta-thoracic discs located on either side in the larvae gives rise to the corresponding fore- and hind-wings of the adult. These discs do not experience massive cell rearrangements during metamorphosis and display the adult wing vein pattern. We have analysed wing development in B. mori by two approaches, viz., expression of patterning genes in larval wing discs, and regulatory capacities of larval discs following explantation or perturbation. Expression of Nubbin is seen all over the presumptive wing blade domains unlike in Drosophila, where it is confined to the hinge and the wing pouch. Excision of meso- and meta-thoracic discs during the larval stages resulted in emergence of adult moths lacking the corresponding wings without any loss of thoracic tissues suggesting independent origin of wing and thoracic primordia. The expression of wingless and distal-less along the dorsal/ventral margin in wing discs correlated well with their expression profile in adult Drosophila wings. Partially excised wing discs did not show in situ regeneration or duplication suggesting their early differentiation. The presence of adult wing vein patterns discernible in larval wing discs and the patterns of marker gene expression as well as the inability of these discs to regulate growth suggested that wing differentiation is achieved early in B. mori. The timings of morphogenetic events are different and the wing discs behave like presumptive wing buds opening out as wing blades in B. mori unlike evagination of only the pouch region as wing blades seen in Drosophila.

  20. Role of Bmznf-2, a Bombyx mori CCCH zinc finger gene, in masculinisation and differential splicing of Bmtra-2.

    Science.gov (United States)

    Gopinath, Gajula; Arunkumar, Kallare P; Mita, Kazuei; Nagaraju, Javaregowda

    2016-08-01

    Deciphering the regulatory factors involved in Bombyx mori sex determination has been a puzzle, challenging researchers for nearly a century now. The pre-mRNA of B. mori doublesex (Bmdsx), a master regulator gene of sexual differentiation, is differentially spliced, producing Bmdsxm and Bmdsxf transcripts in males and females respectively. The putative proteins encoded by these differential transcripts orchestrate antagonistic functions, which lead to sexual differentiation. A recent study in B. mori illustrated the role of a W-derived fem piRNA in conferring femaleness. In females, the fem piRNA was shown to suppress the activity of a Z-linked CCCH type zinc finger (znf) gene, Masculiniser (masc), which indirectly promotes the Bmdsxm type of splicing. In this study, we report a novel autosomal (Chr 25) CCCH type znf motif encoding gene Bmznf-2 as one of the potential factors in the Bmdsx sex specific differential splicing, and we also provide insights into its role in the alternative splicing of Bmtra2 by using ovary derived BmN cells. Over-expression of Bmznf-2 induced Bmdsxm type of splicing (masculinisation) with a correspondingly reduced expression of Bmdsxf type isoform in BmN cells. Further, the site-directed mutational studies targeting the tandem CCCH znf motifs revealed their indispensability in the observed phenotype of masculinisation. Additionally, the dual luciferase assays in BmN cells using 5' UTR region of the Bmznf-2 strongly implied the existence of a translational repression over this gene. From these findings, we propose Bmznf-2 to be one of the potential factors of masculinisation similar to Masc. From the growing number of Bmdsx splicing regulators, we assume that the sex determination cascade of B. mori is quite intricate in nature; hence, it has to be further investigated for its comprehensive understanding. PMID:27260399

  1. Expression of a sugar clade gustatory receptor, BmGr6, in the oral sensory organs, midgut, and central nervous system of larvae of the silkworm Bombyx mori.

    Science.gov (United States)

    Mang, Dingze; Shu, Min; Endo, Haruka; Yoshizawa, Yasutaka; Nagata, Shinji; Kikuta, Shingo; Sato, Ryoichi

    2016-03-01

    Insects taste nonvolatile chemicals through gustatory receptors (Grs) and make choices for feeding, mating, and oviposition. To date, genome projects have identified 69 Gr genes in the silkworm, Bombyx mori; however, the expression sites of these Grs remain to be explored. In this study, we used reverse transcription (RT)-PCR to investigate expression of the B. mori Gr-6 (BmGr6) gene, a member of the putative sugar clade gene family in various tissues. BmGr6 is expressed in the midgut, central nervous system (CNS), and oral sensory organs. Moreover, immunohistochemistry using an anti-BmGr6 antiserum demonstrated that BmGr6 is expressed in cells by oral sensory organs, midgut and nervous system. Furthermore, double-immunohistochemistry indicated that BmGr6 is expressed in midgut enteroendocrine cells, also in CNS neurosecretory cells. In particular, a portion of BmGr6-expressing cells, in both midgut and CNS, secretes FMRFamide-related peptides (FaRPs). These results suggest that BmGr6 functions not only as a taste receptor, but also as a chemical sensor such as for the regulation of gut movement, physiological conditions, and feeding behavior of larvae. PMID:26721200

  2. Influência das dietas artificiais e folhas in natura no ganho de peso das larvas do bicho-da-seda (Bombyx mori L. Influence of artificial diets and natural leaves in weight gain of silkworm larvae (Bombyx mori L.

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    Adriana Evangelista Rodrigues

    2000-05-01

    Full Text Available A utilização de dietas artificiais no Japão já é uma realidade e no Brasil iniciam-se os estudos, buscando encontrar um balanceamento que satisfaça as necessidades das larvas do bicho-da-seda (Bombyx mori L.. Desta forma, foram testados dois cultivares de amoreira como ingredientes das dietas, avaliando-se a qualidade do alimento através do ganho médio de peso das larvas. As larvas foram separadas em parcelas no início do 1º instar, quando já começaram a receber as dietas artificiais e as folhas in natura. Para o cálculo do ganho médio de peso analisou-se o peso inicial e final das larvas a cada instar. Quando as larvas receberam dietas artificiais no 1º e 2º instares e folhas in natura a partir do 3º instar não apresentaram diferenças significativas, sugerindo a utilização de dietas em criadeiras nas Fiações.The use of artificial diets in Japan is a well known fact. In Brazil experiments are assayed to find a balanced diet to meet the needs of the silkworm (Bombyx mori L.. Two mulberry cultivars were tested in diets to evaluate the quality of food by average gain of weight in larvae. Larvae were first separated in the first instar and fed with diets and natural leaves. As a rough estimate of mean gain of weight, initial and final weights in each instar were recorded. When the silkworms were fed with artificial diets in the first and second instars and with natural leaves in the third, fourth and fifth instars no significant differences were recorded. The use of diets in the first instars at the silk industry may be thus recommended.

  3. Fibroin silk proteins from the nonmulberry silkworm Philosamia ricini are biochemically and immunochemically distinct from those of the mulberry silkworm Bombyx mori.

    Science.gov (United States)

    Ahmad, Raies; Kamra, Anita; Hasnain, Seyed Ehtesham

    2004-03-01

    Silk proteins were isolated from the cocoons of the nonmulberry silkworm, Philosamia ricini. Three polypeptides of 97, 66, and 45 kDa were identified. The 66-kDa molecule represented sericin, whereas the 97-kDa and the 45-kDa polypeptides linked together through a disulfide bond constituted the fibroin protein. Antibodies raised against the 97-kDa P. ricini fibroin heavy chain reacted specifically with this molecule and did not recognize fibroin heavy chain from another nonmulberry silkworm, Antheraea assama or from the mulberry silkworm, Bombyx mori, suggesting the presence of P. ricini species-specific determinants in this heavy chain. Antibodies generated against fibroin light chain of P. ricini also showed similar reactivity pattern. Immunoblot analysis with proteins isolated from the silk glands of P. ricini at different stages of larval development showed that the expression of fibroin heavy chain was developmentally and spatially regulated. The protein was most abundant in the 5th instar larva, and could be detected in the middle and the posterior but not the anterior silk glands. The amino acid composition of the 97-kDa fibroin protein showed abundance of glutamic acid and did not contain (Gly-Ala)(n) motifs, a characteristic feature of B. mori fibroin heavy chain. Our study reveals significant differences between the nonmulberry silkworm P. ricini and the mulberry silkworm B. mori in the biochemical composition and immunochemical characteristics of fibroin heavy chain. These differences might be responsible for the differences seen in the quality of silk produced by these two silkworms. PMID:15068584

  4. Efficient large-scale protein production of larvae and pupae of silkworm by Bombyx mori nuclear polyhedrosis virus bacmid system

    International Nuclear Information System (INIS)

    Silkworm is one of the most attractive hosts for large-scale production of eukaryotic proteins as well as recombinant baculoviruses for gene transfer to mammalian cells. The bacmid system of Autographa californica nuclear polyhedrosis virus (AcNPV) has already been established and widely used. However, the AcNPV does not have a potential to infect silkworm. We developed the first practical Bombyx mori nuclear polyhedrosis virus bacmid system directly applicable for the protein expression of silkworm. By using this system, the green fluorescence protein was successfully expressed in silkworm larvae and pupae not only by infection of its recombinant virus but also by direct injection of its bacmid DNA. This method provides the rapid protein production in silkworm as long as 10 days, is free from biohazard, thus will be a powerful tool for the future production factory of recombinant eukaryotic proteins and baculoviruses

  5. Chromosomal localization of silkworm (Bombyx mori) sericin gene 1 and chymotrypsin inhibitor 13 using fluorescence in situ hybridization

    Institute of Scientific and Technical Information of China (English)

    Yutaka; BANNO; Hiroshi; FUJII

    2008-01-01

    The chromosomal locations of two single-copy genes, Ser-1 and CI-13, in silkworm (Bombyx mori) were detected at the molecular cytogenetics level by fluorescence in situ hybridization in the study. The results showed that Ser-1 is located near the distal end of the 11th linkage group, relatively at the 12.5±1.4 position in pachytene; and that CI-13 has been mapped near the distal end of the 2nd linkage group, relatively at the 8.2±1.2 position in pachytene. Furthermore, their location model map-FISH map on silkworm chromosome was drawn. The FISH technique and its application to silkworm are also discussed in this paper.

  6. High-level expression of human butyrylcholinesterase gene in Bombyx mori and biochemical-pharmacological characteristic study of its product.

    Science.gov (United States)

    Wei, W L; Qin, J C; Sun, M J

    2000-07-01

    The human butyrylcholinesterase (BChE, EC 3.1.1.8) gene was highly expressed in Bombyx mori using baculovirus vector, and the biochemical-pharmacological properties of its product were studied. BChE cDNA was cloned into transfer vector pBn96 and co-transfected with wild-type Bombyx mori nucleopolyhedrovirus (BmNPV) DNA into BmN cells. The recombinant virus with the highest enzyme activity was sorted out and purified. Once the BmN cells or silkworm larvae had been infected with the recombinant virus, recombinant human BChE (rhBChE) could be secreted into the culture medium or the hemolymph of the larvae at levels of 1.5 mg x L(-1) and 35 mg x L(-1), respectively. Western blot and enzymatic staining of the electrophoresis gel of non-denatured protein showed that rhBChE manifested similar antigenicity and enzyme activity to native human BChE (nhBChE). The production of rhBChE in the hemolymph was 23-fold higher than that in BmN cells and about 280-fold that in Chinese hamster overy cells (125 microg x L(-1)). This is the first report of human BChE expression in silkworm with the highest level of yield so far. rhBChE was highly similar to nhBChE in respect to substrate affinity, inhibitor sensitivity, and reactivity of the inhibited enzyme. It is suggested that rhBChE functions as well as nhBChE and has potential practical value. PMID:10807953

  7. Molecular cloning and functional characterization of the dual oxidase (BmDuox gene from the silkworm Bombyx mori.

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    Xiaolong Hu

    Full Text Available Reactive oxygen species (ROS from nicotinamide adenine dinucleotide phosphate (NADPH oxidases and their related dual oxidases are known to have significant roles in innate immunity and cell proliferation. In this study, the 5,545 bp cDNA of the silkworm Bombyx mori dual oxidase (BmDuox gene containing a full-length open reading frame was cloned. It was shown to include an N-terminal signal peptide consisting of 28 amino acid residues, a 240 bp 5'-terminal untranslated region (5'-UTR, an 802 bp 3'-terminal region (3'-UTR, which contains nine ATTTA motifs, and a 4,503 bp open reading frame encoding a polypeptide of 1,500 amino acid residues. Structural analysis indicated that BmDuox contains a typical peroxidase domain at the N-terminus followed by a calcium-binding domain, a ferric-reducing domain, six transmembrane regions and binding domains for flavin adenine dinucleotide (FAD and nicotinamide adenine dinucleotide (NAD. Transcriptional analysis revealed that BmDuox mRNA was expressed more highly in the head, testis and trachea compared to the midgut, hemocyte, Malpighian tube, ovary, fat bodies and silk glands. BmDuox mRNA was expressed during all the developmental stages of the silkworm. Subcellular localization revealed that BmDoux was present mainly in the periphery of the cells. Some cytoplasmic staining was detected, with rare signals in the nucleus. Expression of BmDuox was induced significantly in the larval midgut upon challenge by Escherichia coli and Bombyx mori nucleopolyhedrovirus (BmNPV. BmDuox-deleted larvae showed a marked increase in microbial proliferation in the midgut after ingestion of fluorescence-labeled bacteria compared to the control. We conclude that reducing BmDuox expression greatly increased the bacterial load, suggesting BmDuox has an important role in inhibiting microbial proliferation and the maintenance of homeostasis in the silkworm midgut.

  8. Genome-wide identification and characterization of ATP-binding cassette transporters in the silkworm, Bombyx mori

    Directory of Open Access Journals (Sweden)

    Zhang Jianzhen

    2011-10-01

    Full Text Available Abstract Background The ATP-binding cassette (ABC transporter superfamily is the largest transporter gene family responsible for transporting specific molecules across lipid membranes in all living organisms. In insects, ABC transporters not only have important functions in molecule transport, but also play roles in insecticide resistance, metabolism and development. Results From the genome of the silkworm, Bombyx mori, we have identified 51 putative ABC genes which are classified into eight subfamilies (A-H by phylogenetic analysis. Gene duplication is very evident in the ABCC and ABCG subfamilies, whereas gene numbers and structures are well conserved in the ABCD, ABCE, ABCF, and ABCH subfamilies. Microarray analysis revealed that expression of 32 silkworm ABC genes can be detected in at least one tissue during different developmental stages, and the expression patterns of some of them were confirmed by quantitative real-time PCR. A large number of ABC genes were highly expressed in the testis compared to other tissues. One of the ABCG genes, BmABC002712, was exclusively and abundantly expressed in the Malpighian tubule implying that BmABC002712 plays a tissue-specific role. At least 5 ABCG genes, including BmABC005226, BmABC005203, BmABC005202, BmABC010555, and BmABC010557, were preferentially expressed in the midgut, showing similar developmental expression profiles to those of 20-hydroxyecdysone (20E-response genes. 20E treatment induced the expression of these ABCG genes in the midgut and RNA interference-mediated knockdown of USP, a component of the 20E receptor, decreased their expression, indicating that these midgut-specific ABCG genes are 20E-responsive. Conclusion In this study, a genome-wide analysis of the silkworm ABC transporters has been conducted. A comparison of ABC transporters from 5 insect species provides an overview of this vital gene superfamily in insects. Moreover, tissue- and stage-specific expression data of the

  9. Gender Influenced Spore Dimorphism in Nosema bombycis Nageli Causing Pebrine Disease in Mulberry Silkworm, Bombyx mori L.

    Directory of Open Access Journals (Sweden)

    Satadal CHAKRABARTY

    2013-04-01

    Full Text Available Nosema bombycis is a pathogen causing pebrine disease of the mulberry silkworm, Bombyx mori. The disease spreads mainly through transovarian transmission of environmental spore and secondarily through contaminated food, rearing appliances, etc. by primary spores. Ultra-structure studies using Scanning and Transmission Electron Microscopy of the two spores revealed differences in the primary spore which contained a Short Polar Tube (ST with a thin wall (< 200 nm, and the environmental spore which contained a Long Polar Tube (LT with a thick wall (> 200 nm. It is observed that the yield of spore with LT is highest in female moths, whereas, it is spores with ST are highest in male moths. Besides ultra-structures, the development pattern of the two types of spores is also different. It is an interesting finding in the present study that, spores of N. bombycis produced two types of spores and multiplied in different gender under the influence of the host’s reproductive role and physiology for transmission of disease. The detailed study on ultra-structure of disporous N. bombycis in both the sexes of B. mori along with their development in the life cycle stages of silkworms with special reference to the inoculum concentration of spore is discussed.  

  10. The C-terminus of DSX(F5) protein acts as a novel regulatory domain in Bombyx mori.

    Science.gov (United States)

    Duan, Jianping; Meng, Xianxin; Ma, Sanyuan; Wang, Feng; Guo, Huozhen; Zhang, Liying; Zhao, Ping; Kan, Yunchao; Yao, Lunguang; Xia, Qingyou

    2016-08-01

    The doublesex gene regulates the somatic sexual development of Bombyx mori by alternatively splicing into sex-specific splice forms. In our previous study, the splice form Bmdsx (F7) , which encodes the BmDSX(F5) protein, was found to be expressed in a female-specific manner and to contain a novel C-terminus. In this study, we aimed to investigate the role of this C-terminus. Two transgenic lines, L1 and L2, were constructed to ectopically express Bmdsx (F7) in males. Phenotype and W chromosome-specific polymerase chain reaction (PCR) analysis showed that developmental abnormalities and sex reversal did not occur. Moreover, the sex ratio was also normal. Quantitative PCR revealed that the expression levels of SP1 and Vg were upregulated in the fat body of transgenic males. Additionally, the expression level of PBP was downregulated in the antenna of transgenic males. The results suggested that the C-terminus of BmDSX(F5) functioned as a regulatory domain during regulation of downstream target gene expression and that BmDSX(F5) participated in the sexual development of somatic cells together with other DSX proteins in B. mori. PMID:26975733

  11. Construction of an in vivo System for Functional Analysis of the Genes Involved in Sex Pheromone Production in the Silkmoth, Bombyx mori

    OpenAIRE

    Ken-IchiMoto

    2012-01-01

    Moths produce species-specific sex pheromones to attract conspecific mates. The biochemical processes that comprise sex pheromone biosynthesis are precisely regulated and a number of gene products are involved in this biosynthesis and regulation. In recent years, at least 300 EST clones have been isolated from Bombyx mori pheromone gland (PG) specific cDNA libraries with some of those clones (i.e., B. mori PG-specific desaturase1 (Bmpgdesat1), PG-specific fatty-acyl reductase (pgFAR), PG-spec...

  12. Folhas de amoreira tratadas com vitamina e esterol e seus efeitos em parâmetros biológicos de Bombyx mori L.

    OpenAIRE

    Sergio Antonio De Bortoli; Roque Takahashi; José Ednilson Miranda; Luciane Sandrini Dias; Caroline Placidi De Bortoli; Sergio Leandro Placidi De Bortoli

    2012-01-01

    O objetivo do trabalho foi avaliar o efeito dos aditivos foliares Decanoato de nandrolona, Ácido ascórbico, Palmitato de retinol e Acetato de retinol, em diferentes concentrações, sobre alguns parâmetros biológicos do bicho da seda, Bombyx mori. Verificou-se que o ácido ascórbico e a nandrolona, ambos a 0,5%, proporcionam os melhores valores para o desenvolvimento corpóreo das lagartas de B. mori, valores estes que não correspondem a incrementos significativos na produção de seda; 1,0% palmit...

  13. Female sex pheromone and male behavioral responses of the bombycid moth Trilocha varians: comparison with those of the domesticated silkmoth Bombyx mori

    Science.gov (United States)

    Daimon, Takaaki; Fujii, Takeshi; Yago, Masaya; Hsu, Yu-Feng; Nakajima, Yumiko; Fujii, Tsuguru; Katsuma, Susumu; Ishikawa, Yukio; Shimada, Toru

    2012-03-01

    Analysis of female sex pheromone components and subsequent field trap experiments demonstrated that the bombycid moth Trilocha varians uses a mixture of ( E, Z)-10,12-hexadecadienal (bombykal) and ( E,Z)-10,12-hexadecadienyl acetate (bombykyl acetate) as a sex pheromone. Both of these components are derivatives of ( E,Z)-10,12-hexadecadienol (bombykol), the sex pheromone of the domesticated silkmoth Bombyx mori. This finding prompted us to compare the antennal and behavioral responses of T. varians and B. mori to bombykol, bombykal, and bombykyl acetate in detail. The antennae of T. varians males responded to bombykal and bombykyl acetate but not to bombykol, and males were attracted only when lures contained both bombykal and bombykyl acetate. In contrast, the antennae of B. mori males responded to all the three components. Behavioral analysis showed that B. mori males responded to neither bombykal nor bombykyl acetate. Meanwhile, the wing fluttering response of B. mori males to bombykol was strongly inhibited by bombykal and bombykyl acetate, thereby indicating that bombykal and bombykyl acetate act as behavioral antagonists for B. mori males. T. varians would serve as a reference species for B. mori in future investigations into the molecular mechanisms underlying the evolution of sex pheromone communication systems in bombycid moths.

  14. EFFECTS OF JINLU, AN ANTI-JUVENILE HORMONE ON THE GROWTH, ULTRA-STRUCTURE OF THE CORPORA ALLATA AND PROTHORACIC GLAND OF SILKWORM, BOMBYX MORI L

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    The 4th instar larvae of the silkworm, Bombyx mori L, when treated with anti-juvenile hormone (Jinlu) had its larval period extended by 2 days and the total larval period shortened by about 4 days. The conversion ratio of tetramolters into trimolters was 100%. But anti-juvenile hormone administration to the 5th instar larvae lengthened the silkworm age by one day. When anti-juvenile hormone was administered, we could find many neurosecretory granules of the brain transferred to the cells of the corpora allata, but there was little endoplasmic reticulum. In the prothoracic gland, the micropile edge was clear and there were large nucleoli, mitochondria and endoplasmic reticulum. This study was carried out to show that anti-JH compound inhibits the secretion of Juvenile hormone in silkworm Bombyx mori L. The investigation revealed that the anti-juvenile hormone inhibited the secretion of corpora allata and initiated the activity of the prothoracic gland.

  15. Development of a standard acute dietary toxicity test for the silkworm (Bombyx mori L.)

    NARCIS (Netherlands)

    Sun, X.; Valk, H.; Jiang, H.; Wang, X.; Yuan, S.; Zhang, Y.; Roessink, I.; Gao, X.

    2012-01-01

    Larvae of the silkworm (Bombyx mod L.) may be exposed to pesticide residues on the leaves of their food plant, the mulberry tree (Morus spp.), which can lead to adverse effects on silk production. A new acute dietary toxicity test method was evaluated as the basis for pesticide risk assessment. A se

  16. Identification of conserved microRNAs in Bombyx mori (silkworm) and regulation of fibroin L chain production by microRNAs in heterologous system.

    Science.gov (United States)

    Cao, Jun; Tong, Chuanzhou; Wu, Xiaojie; Lv, Jianning; Yang, Zhaolin; Jin, Yongfeng

    2008-12-01

    MicroRNAs (miRNAs) are an extensive class of small non-coding RNAs with important roles in the regulation of gene expression. Little study has been performed on identifying miRNAs in Bombyx mori (silkworm). Here, 41 conserved miRNAs were identified by a computational homology search approach and some of them were selected and confirmed experimentally. Some characteristics such as position-specific nucleotide distribution, evolutionary conservation and clustered organization have been described. Through general method of miRNA target identification, we also find that at least 4 miRNAs (miR-33/-190/-276/-7) target fibroin L chain transcripts, suggesting that miRNAs might play an important role in the regulation of silk protein production. In order to validate this, 3'-UTR of fibroin L chain mRNA was cloned and several expression vectors were constructed. Assays of semi-RT-PCR, histochemical and biochemical assays after agroinfiltration indicated that miRNAs can repress reporter gene (GUS) expression with sequence complement. Clearly, agroinfiltration proved a highly efficient method to identify the function of miRNAs inhibiting their targets mRNA in heterologous system. PMID:18977439

  17. Phantom encodes the 25-hydroxylase of Drosophila melanogaster and Bombyx mori: a P450 enzyme critical in ecdysone biosynthesis.

    Science.gov (United States)

    Warren, James T; Petryk, Anna; Marqués, Guillermo; Parvy, Jean-Philippe; Shinoda, Tetsuro; Itoyama, Kyo; Kobayashi, Jun; Jarcho, Michael; Li, Yutai; O'Connor, Michael B; Dauphin-Villemant, Chantal; Gilbert, Lawrence I

    2004-09-01

    We have reported recently the identification and characterization of the last three mitochondrial cytochrome P450 enzymes (CYP) controlling the biosynthesis of 20-hydroxyecdysone, the molting hormone of insects. These are encoded by the following genes: disembodied (dib, Cyp302a1, the 22-hydroxylase); shadow (sad, Cyp315a1, the 2-hydroxylase); and shade (shd, Cyp314a1, the 20-hydroxylase). Employing similar gene identification and transfection techniques and subsequent biochemical analysis of the expressed enzymatic activity, we report the identity of the Drosophila gene phantom (phm), located at 17D1 of the X chromosome, as encoding the microsomal 25-hydroxylase (Cyp306a1). Similar analysis following differential display-based gene identification has also resulted in the characterization of the corresponding 25-hydroxylase gene in Bombyx mori. Confirmation of 2,22,25-trideoxyecdysone (3beta,5beta-ketodiol) conversion to 2,22-dideoxyecdysone (3beta,5beta-ketotriol) mediated by either Phm enzyme employed LC, MS and definitive NMR analysis. In situ developmental gene analysis, in addition to northern, western and RT-PCR techniques during Drosophila embryonic, larval and adult development, are consistent with this identification. That is, strong expression of phm is restricted to the prothoracic gland cells of the Drosophila larval ring gland, where it undergoes dramatic changes in expression, and in the adult ovary, but also in the embryonic epidermis. During the last larval-larval transition in Bombyx, a similar expression pattern in the prothoracic gland is observed, but as in Drosophila, slight expression is also present in other tissues, suggesting a possible additional role for the phantom enzyme. PMID:15350618

  18. Intracellular signal transduction of PBAN action in the silkworm, Bombyx mori: involvement of acyl CoA reductase.

    Science.gov (United States)

    Ozawa, R; Matsumoto, S

    1996-03-01

    In the silkworm, Bombyx mori, production of the sex pheromone bombykol is regulated by a neurohormone termed PBAN. We have detected the activity of acyl CoA reductase in the pheromone gland of B. mori by using palmitoyl CoA as a substrate. The acyl CoA reductase requires NADPH, but not NADH, as a proton dono. When the pheromone gland was incubated with the PBAN fragment peptide TKYFSPRLamide, palmitoyl CoA was incorporated and converted into the corresponding C16 alcohols. Radio HPLC analysis revealed that these C16 alcohols were hexadecan-1-ol (81.2%), (Z)-11-hexadecen-1-ol (12.3%), and (E, Z)-10, 12-hexadecadien-1-ol (= bombykol, 6.5%). The production of C16 alcohols in the pheromone gland was inhibited by the known bombykol biosynthesis inhibitors EDTA, LaCl3, W-7, trifluoperazine, p-nitrophenyl phosphate, NaF and compactin. By contrast, when the pheromone gland homogenate was incubated in the presence of palmitoyl CoA and NADPH, production of C16 alcohols was affected by compactin, W-7 and trifluoperazine, but not by EDTA, LaCl3, p-nitrophenyl phosphate and NaF. These results indicate that compactin, W-7 and trifluoperazine directly suppress the step catalyzed by acyl CoA reductase, whereas EDTA, LaCl3, pNPP, and NaF inhibit bombykol production by affecting other biochemical steps in the signal transduction of PBAN action. The present results also imply that PBAN regulates the step catalyzed by acyl CoA reductase and that palmitoyl CoA could be used as a substrate of the acyl CoA reductase that regulates bombykol biosynthesis. PMID:8900596

  19. Gamma radiation effects of {sup 60} Co on Bombyx mori (Lep., Bombycidae) modifying the silk fiber production; Influencia da radiacao gama ({sup 60} Co) na producao de fios de seda em Bombyx mori(Lep.,Bombycidae)

    Energy Technology Data Exchange (ETDEWEB)

    Carneiro Junior, Francisco [Universidade Metodista de Piracicaba (UNIMEP), SP (Brazil); Bendassolli, Jose A. [Centro de Energia Nuclear na Agricultura (CENA), Piracicaba, SP (Brazil)

    1997-12-01

    The present work aimed to verify the biological effects of the application of different doses of gamma radiation during the fifth instar of the silkworm catepillar. Sevently eight silkworm caterpillars (Bombyx mori) were irradiated with {gamma}{sup 60} Co radiation at the initial period of the fifth instar. The caterpillars were divided and classified in six batches of thirteen individuals each. Treatments 1 through 5 received 20, 40, 60, 80 and 100 Gy, respectively, and the control, also consisted of thirteen caterpillars, was not irradiated. The results showed a general increase in the silk fiber content in the irradiated batches compared to the control. The weight of the silk cocoons was higher with increasing doses of irradiation, from 20 to 80 Gy, respectively, followed by a decrease in weight in the treatment irradiated with 100 Gy. the results obtained in this experiment enable the conclusion that the radiation applied to the caterpillars significantly influenced the production of silk fiber in this species. (author). 4 refs., 2 figs., 3 tabs.

  20. Molecular cloning, expression and identification of the promoter regulatory region for the neuropeptide trissin in the nervous system of the silkmoth Bombyx mori.

    Science.gov (United States)

    Roller, Ladislav; Čižmár, Daniel; Gáliková, Zuzana; Bednár, Branislav; Daubnerová, Ivana; Žitňan, Dušan

    2016-06-01

    Trissin has recently been identified as a conserved insect neuropeptide, but its cellular expression and function is unknown. We detected the presence of this neuropeptide in the silkworm Bombyx mori using in silico search and molecular cloning. In situ hybridisation was used to examine trissin expression in the entire central nervous system (CNS) and gut of larvae, pupae and adults. Surprisingly, its expression is restricted to only two pairs of small protocerebral interneurons and four to five large neurons in the frontal ganglion (FG). These neurons were further characterised by subsequent multiple staining with selected antibodies against insect neuropeptides. The brain interneurons innervate edges of the mushroom bodies and co-express trissin with myoinhibitory peptides (MIP) and CRF-like diuretic hormones (CRF-DH). In the FG, one pair of neurons co-express trissin with calcitonin-like diuretic hormone (CT-DH), short neuropeptide F (sNPF) and MIP. These neurons innervate the brain tritocerebrum and musculature of the anterior midgut. The other pair of trissin neurons in the FG co-express sNPF and project axons to the tritocerebrum and midgut. We also used the baculovirus expression system to identify the promoter regulatory region of the trissin gene for targeted expression of various molecular markers in these neurons. Dominant expression of trissin in the FG indicates its possible role in the regulation of foregut-midgut contractions and food intake. PMID:26809512

  1. Impact of heat shock on heat shock proteins expression,biological and commercial traits of Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    VASUDHA B. CHAVADI; APARNA H. S OSALEGOWDA; MANJUNATHA H.B OREGOWDA

    2006-01-01

    We report the thermotolerance of new bivoltine silkworm, Bombyx mori strains NB4D2, KSO1, NP2, CSR2 and CSR4 and differential expression of heat shock proteins at different instars. Different instars of silkworm larva were subjected to heat shock at 35℃,40℃ and 45℃ for 2 hours followed by 2 hours recovery. Heat shock proteins were analyzed by SDS-PAGE. The impact of heat shock on commercial traits of cocoons was analyzed by following different strategies in terms of acquired thermotolerance over control. Comparatively NP2 exhibited better survivability than other strains. Resistance to heat shock was increased as larval development proceeds in the order of first instar > second instar > third instar > fourth instar > fifth instar in all silkworm strains. Expression of heat shock proteins varies in different instars. 90 kDa in the first, second and third instars, 84 kDa in the fourth instar and 84, 62, 60, 47 and 33 kDa heat shock proteins in fifth instar was observed in response to heat shock. Relative influence of heat shock on commercial traits that correspond to different stages was significant in all strains. In NB4D2, cocoon and shell weight significantly increased to 17.52% and 19.44% over control respectively. Heat shock proteins as molecular markers for evaluation and evolution of thermotolerant silkworm strains for tropics was discussed.

  2. Analysis of genetic relationship in mutant silkworm strains of Bombyx mori using inter simple sequence repeat (ISSR) markers

    Institute of Scientific and Technical Information of China (English)

    Dhanikachalam Velu; Kangayam M. Ponnuvel; Murugiah Muthulakshmi; Randhir K. Sinha; Syed M.H. Qadri

    2008-01-01

    Amplified inter simple sequence repeats (ISSR) markers were used to determine genetic relationships among mutant silkworm strains of Bombyx mori. Fifteen ISSR primers containing simple sequence repeat (SSR) motifs were used in this study. A total of 113 markers were produced among 20 mutant swains, of which 73.45% were found to be polymorphic. In selected mutant genetic stocks, the average number of observed allele was (1.7080±0.4567), effective alleles (1.5194±0.3950) and genetic diversity (Ht) (0.2901±0.0415). The dendrogram produced using the unweighted pair group method with arithmetic means (UPGMA) and cluster analysis made using Nei's genetic distance resulted in the formation of one major group containing 6 groups separated 20 mutant silkworm strains. Therefore, ISSR amplification is a valuable method for determining the genetic variability among mutant silkworm swains. This efficient molecular marker would be useful for characterizing a considerable number of silkworm swains maintained at the germplasm center.

  3. Identification and characterization of the Bombyx mori myosin II essential light chain and its effect in BmNPV infection

    Directory of Open Access Journals (Sweden)

    L Hao

    2015-02-01

    Full Text Available Myosin, as a type of molecular motor, is mainly involved in muscle contraction. Recently, myosin research has made considerable progress. However, the function of Bombyx mori myosin remains unclear. In this study, we cloned the BmMyosin II essential light chain (BmMyosin II ELC gene from a cDNA library of silkworm, which had an open reading frame (ORF of 444 bp encoding 147 amino acids (about 16 kDa. After analyzing their sequences, BmMyosin II ELC was similar to the ELCs of 27 other Myosin II types, which contained EFh domain that bound Ca2+. In addition, 28 sequences had five motifs, motifs 1 and 3 were relatively conserved. We constructed two vectors with BmMyosin to transfect MGC803 or BmN, monolayer wound healing of cells indicated they can promote cell migration successfully. For three fifth instar silkworms, Bm306, BmNB, BmBC8, we mainly analyzed the change of BmMyosin II ELC from transcription and translation after infecting with nucleopolyhedrovirus (BmNPV. We found that gene expression of resistant strains were higher than susceptible strains at 12 h, while the result of the translation level was opposite that of the transcription level. Through in vitro protein interactions, we found BmMyosin II ELC can interact with BmNPV ubiquitin.

  4. Effect of marine extracts on the microbial pathogens causing flacherie in the mulberry silkworm, Bombyx mori L.

    Institute of Scientific and Technical Information of China (English)

    KChairman; AJARanjit Singh; GAmalarani; CPadmalatha; GAlagumuthu

    2012-01-01

    Objective: Silkworms are invertebrate animals that are killed by bacteria pathogenic against humans, such as Staphylococcus aureus, Streptococcus pyogenes, Pseudomonas aeruginosa and Vibrio cholera. Biochemical characterization of the microbes in the haemolymph of diseased silkworm collected during the survey indicated the presence of Bacillus sp., Streptococcus sp., Staphylococcus sp. and Pseudomonas sp. in the culture. Methods: Studies were carried out in vitro to assess the efficacy of some marine extracts for the containment of these microbes through turbidimetry analysis and zone of inhibition test. Results: The observations made during this study revealed that the ethyl acetate crude extracts of two marine samples are Auroraglobostellata and Spirostella inconstans var. moendrina Dendy effective against these microbes causing flacherie diseases in silkworm. The comparison of their effects indicated that ethyl acetate extracts were generally more effective Extensive studies using these extracts on the growth and cocoon production of the mulberry silkworm, Bombyx mori L. are likely to throw much light on the possibility of using such extracts as a prophylactic measure during silkworm rearing to improve silk production. Conclusions: Also, the results indicate that maybe plays a possible role in the contamination of humans and animals, in particular silkworms, while marine extracts showed a potential to control the contamination caused by bacterial diseases.

  5. The effects of BmNPV on biochemical changes in primary cultures of Bombyx mori embryonic tissue.

    Science.gov (United States)

    Matindoost, Leila; Sendi, Jalal Jalali; Soleimanjahi, Hoorieh; Etebari, Kayvan; Rahbarizade, Fateme

    2008-01-01

    The effect of Bombyx mori nuclear polyhedrosis virus (BmNPV) on biochemical changes of TC-100 medium containing 10% fetal bovine serum (FBS) in embryonic primary cultures of silkworm was investigated. The primary cultures that reached 60% confluence were infected by 0.5, 1, and 2-ml viral inoculums (diluted with TC-100 medium representing multiplicity of infection (MOI) of 0.25, 0.5, and 1). Glucose, uric acid, urea, total protein, cholesterol, and alkaline phosphatase were measured in the medium of BmNPV-infected primary cultures. All biochemical compounds showed significant changes. Glucose decreased considerably by about 55 mg/ml, while different concentrations of the virus inoculums did not demonstrate significant differences among them. Total protein had only increased in 2 ml concentration and there were no changes in other concentrations. Uric acid as a by-product accumulated dramatically in all concentrations, while the amount of urea reduced in all treatments and this reduction was more evident in lower concentrations. Cholesterol consumption was high in cultures postinfection, while alkaline phosphatase (ALP) activity decreased in infected cells. PMID:18288542

  6. Characterization of sericin powder prepared from citric acid-degraded sericin polypeptides of the silkworm, Bombyx Mori.

    Science.gov (United States)

    Kurioka, Akira; Kurioka, Fujie; Yamazaki, Masayoshi

    2004-04-01

    Acid-degraded sericin powder (AC-SP) was prepared from aqueous solution containing citric acid-degraded sericin polypeptides of Bombyx mori. The morphological and biochemical properties of AC-SP were compared with those of alkali-degraded sericin powder (AL-SP) and hot-water degraded sericin powder (HW-SP). Based on an SEM analysis, AC-SP showed a thin film structure of 10-100 microm with good dispersity while AL-SP and HW-SP had a much larger thin film structure (<500 microm). The extract of AC-SP showed stronger trypsin inhibitor activity due to cocoon shell trypsin inhibitor (CSTI-IV) than that of HW-SP. The extract of AL-SP showed no CSTI-IV activity. It was found that AC-SP was a trypsin inhibitor complex powder and that the release of CSTI-IV from AC-SP depended on pH and ion strength. Similar powder materials were obtained when such organic acids as tartaric acid and succinic acid were used. These results suggest that the acid-degraded sericin polypeptides work as a protein matrix to which CSTI-IV may bind ionically. PMID:15118302

  7. Juvenile hormone diol kinase, a calcium-binding protein with kinase activity, from the silkworm, Bombyx mori.

    Science.gov (United States)

    Li, Sheng; Zhang, Qi-Rui; Xu, Wei-Hua; Schooley, David A

    2005-11-01

    Juvenile hormone (JH) diol kinase (JHDK) is an important enzyme involved in the JH degradation pathway. Bombyx mori (Bommo)-JHDK cDNA (637bp) contains an open reading frame encoding a 183-amino acid protein, which reveals a high degree of identity to the two previously reported JHDKs. JHDK is similar to GTP-binding proteins with three conserved sequence elements involved in purine nucleotide binding, contains eight alpha-helices and three EF-hand motifs, and resembles the three-dimensional model of 2SCP and some other calcium-binding proteins. The Bommo-JHDK gene has only a single copy in the silkworm haploid genome, contains only one exon, and its 5'-upstream sequence does not have a JH response element. Although Bommo-JHDK is highly expressed in the gut of the silkworm, its mRNA expression remains at a constant level during larval development suggesting this enzyme is constitutive and not regulated by JH, at least at the transcriptional level. Recombinant Bommo-JHDK catalyzed the conversion of 10S-JH diol into JH diol phosphate, confirming its enzymatic function. Recombinant enzyme formed a dimer and had biochemical characteristics similar to other JHDKs. Bommo-JHDK, a calcium-binding protein with kinase activity, provides unique insights on how JH levels are regulated in the silkworm. PMID:16203205

  8. Release of ecdysteroid-phosphates from egg yolk granules and their dephosphorylation during early embryonic development in silkworm, Bombyx mori.

    Science.gov (United States)

    Yamada, Ryouichi; Yamahama, Yumi; Sonobe, Haruyuki

    2005-02-01

    Newly laid eggs of many insect species store maternal ecdysteroids as physiologically inactive phosphoric esters. In the silkworm Bombyx mori, we previously reported the presence of a specific enzyme, called ecdysteroid-phosphate phosphatase (EPPase), which catalyzes the dephosphorylation of ecdysteroid-phosphates to increase the amount of free ecdysteroids during early embryonic development. In this study, we demonstrated that (1) EPPase is found in the cytosol of yolk cells, (2) ecdysteroid-phosphates are localized in yolk granules, being bound to the yolk protein vitellin (Vn), and (3) Vn-bound ecdysteroid-phosphates are scarcely hydrolyzed by EPPase, although free ecdysteroid-phosphates are completely hydrolyzed by EPPase. Thus, we investigated the mechanism by which ecdysteroid-phosphates dissociate from the Vn-ecdysteroid-phosphate complex, and indicated that the acidification of yolk granules causes the dissociation of ecdysteroid-phosphates from the Vn-ecdysteroid-phosphate complex and thereby ecdysteroid-phosphates are released from yolk granules into the cytosol. Indeed, the presence of vacuolar-type proton-translocating ATPase in the membrane fraction of yolk granules was also verified by Western blot analysis. Our experiments revealed that Vn functions as a reservoir of maternal ovarian ecdysteroid-phosphates as well as a nutritional source during embryonic development. This is the first report showing the biochemical mechanism by which maternal Vn-bound ecdysteroid-phosphates function during early embryonic development. PMID:15738639

  9. Bombyx mori nucleopolyhedrovirus orf8 encodes a nucleic acid binding protein that colocalizes with IE1 during infection.

    Science.gov (United States)

    Imai, N; Kurihara, M; Matsumoto, S; Kang, W-K

    2004-08-01

    This report describes the characterization of the Bombyx mori nucleopolyhedrovirus (BmNPV) orf8 gene. Immunoblot analyses demonstrated that orf8 was expressed as an early gene. The ORF8 protein accumulated in the nucleus, and was maintained at relatively constant levels from 4 to 24 h postinfection. Immunoblot analysis failed to detect ORF8 protein associated with budded virus and occlusion derived virus. In addition, immunohistochemical analysis by confocal microscopy showed that ORF8 protein colocalized with IE1 to specific nuclear foci throughout infection. To further examine the function of ORF8, a reporter gene was inserted into the orf8 reading frame. One orf8 disruption mutant (BmD8), which expressed the N-terminal half of ORF8, was isolated. However, it was not possible to isolate a null mutant, suggesting that orf8 may have an important role during viral infection. Single-step growth curves showed that BV production was reduced in BmD8 infected cells. Biochemical analyses indicated that ORF8 bound to nucleic acids. Together, these results suggest that BmNPV ORF8 may be involved in viral DNA replication and/or transcription. PMID:15290382

  10. The sexual difference in degradation of sulfur amino acids in pupae and adults of the silkworm, Bombyx mori

    International Nuclear Information System (INIS)

    The degradation of sulfur amino acids in pupae and adults of the silkworm, Bombyx mori, was investigated. The rate of 14CO2 production from DL-cystine-1-14C varied during pupal development and also between male and female significantly; in the male the 14CO2 production increased markedly according to pupal development, whereas in the female it was kept at a low level until emergence, then increased slightly. Total sulfur in meconium of the female was less than a half the quantity of that of the male. However, in the ovary-ectomyzed female the amount of total sulfur in meconium and the rate of 14CO2 production were increased, almost similar to the case of the male. It was also confirmed that the 14C of DL-cystine-1-14C was incorporated efficiently into egg protein at the end of pupal stage. These results indicated that the rate of degradation of sulfur amino acids in the pupal stage was much active in male than in female. Such a sexual difference is considered to be related closely to the formation of egg, especially egg-shell. (auth.)

  11. SSR based linkage and mapping analysis of C, a yellow cocoon gene in the silkworm, Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    Yun-Po Zhao; Mu-Wang Li; An-Ying Xu; Cheng-Xiang Hou; Ming-Hui Li; Qiu-Hong Guo; Yong-Ping Huang; Xi-Jie Guo

    2008-01-01

    The yellow color of the cocoon of the silkworm Bombyx mori is controlled by three genes, Y ( Yellow haemolymph ), I (Yellow inhibitor) and C (Outer-layer yellow cocoon),which are located on linkage groups 2, 9 and 12, respectively. Taking advantage of a lack of crossing over in females, reciprocal backcrossed F1 (BC1) progeny were used for linkage analysis and mapping of the C gene using silkworm strains C108 and KY, which spin white and yellow cocoons, respectively. DNA was extracted from individual pupae and analyzed for simple sequence repeat (SSR) markers. The C gene was found to be linked to seven SSR markers. All the yellow cocoon individuals from a female heterozygous backcross (BC1F) showed a heterozygous profile for SSR markers on linkage group 12,whereas individuals with light yellow cocoons showed the homozygous profile of the strain C 108. Using a reciprocal heterozygous male backcross (BC1M), we constructed a linkage map of 36.4 cM with the C gene located at the distal end, and the closest SSR marker at a distance of 13.9 cM.

  12. Isolation and characterization of lipase-producing bacteria in the intestine of the silkworm, Bombyx mori, reared on different forage.

    Science.gov (United States)

    Feng, Wei; Wang, Xiao-Qiang; Zhou, Wei; Liu, Guang-Ying; Wan, Yong-Ji

    2011-01-01

    The silkworm, Bombyx mori L. (Lepidoptera: Bombycidae), an oligophagous insect that mainly feeds on mulberry leaves, is susceptible to entomopathogen infection when reared with tricuspid cudrania leaves. A total of 56 dominant bacterial strains, classified into 12 phylotypes based on bacteriological properties and analysis of 16S rRNA genes, were isolated from the intestine of the fourth and fifth instar silkworm larvae. Ten and seven phylotypes exist in the intestine of the silkworm larvae reared with mulberry leaves and tricuspid cudrania leaves, respectively. Four of them are common in the intestine of the two treatment groups. By screening their lipolytic ability on a Rhodamine B agar plate, nine lipase-producing bacterial strains were obtained and classified into six genera, including Bacillus, Brevibacterium, Corynebacterium, Staphylococcus, Klebsiella, and Stenotrophomonas. Except for Stenotrophomonas, which is common in both, the other genera only exist in the intestine of the silkworm larvae fed with mulberry leaves. In addition, by culture and fermentation in vitro, the maximum cell density and lipase activity of lipase-producing bacteria were examined at about 48 hours. The results indicate that diet has a significant impact on the gut bacterial community, especially lipase-producing bacteria. We suggest that the difference of lipase-producing bacterial diversity might be related to disease resistance of the silkworm. PMID:22243438

  13. Influence of sericin in alleviating the hydrogen peroxide induced oxidative stress in silkworm Bombyx mori: role of the amino acids

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    AS Micheal

    2014-09-01

    Full Text Available Sericin is an important peptide derived from silk fibre spun by the silkworm Bombyx mori and has various biological activities. The aim of the present study was to characterize the major constituents of sericin that are providing cytoprotective effect against hydrogen peroxide-induced cell damage in midgut epithelial cells and hemocytes of silkworm. Extracted sericin was subjected to LCMS analysis for amino acid composition. Isolated cells of midgut and hemocytes were incubated with sericin or with mixture of serine and aspartic acid prior to suboptimal concentration of hydrogen peroxide treatment. Sericin as well as amino acid mixture reduced the activity of antioxidant enzymes triggered by hydrogen peroxide, inhibited oxidative derivatives such as protein carbonyl and malondialdehyde and increased antioxidant capacity in both the cells studied. Furthermore, sericin and amino acid mixture significantly decreased intracellular reactive oxygen species as assessed by fluorescent detection. These results suggest that major constituent amino acids of sericin defend midgut epithelial cells and hemocytes against oxidative damage by scavenging reactive oxygen species rather than activating antioxidant enzyme system thereby inhibiting cell damage.

  14. The effects of Bombyx mori silk strain and extraction time on the molecular and biological characteristics of sericin.

    Science.gov (United States)

    Siritientong, Tippawan; Bonani, Walter; Motta, Antonella; Migliaresi, Claudio; Aramwit, Pornanong

    2016-01-01

    Sericin was extracted from three strains of Thai Bombyx mori silk cocoons (white shell Chul1/1, greenish shell Chul3/2, and yellow shell Chul4/2) by a high-pressure and high-temperature technique. The characteristics of sericin extracted from different fractions (15, 45, and 60 min extraction process) were compared. No differences in amino acid composition were observed among the three fractions. For all silk strains, sericin extracted from a 15-min process presented the highest molecular weight. The biological potential of the different sericin samples as a bioadditive for 3T3 fibroblast cells was assessed. When comparing sericin extracted from three silk strains, sericin fractions extracted from Chul4/2 improved cell proliferation, while sericin from Chul 1/1 activated Type I collagen production to the highest extent. This study allows the natural variability of sericin obtained from different sources and extraction conditions to be addressed and provides clues for the selection of sericin sources. PMID:26399155

  15. Development of a foot-and-mouth disease virus serotype A empty capsid subunit vaccine using silkworm (Bombyx mori pupae.

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    Zhiyong Li

    Full Text Available Foot-and-mouth disease (FMD is a highly contagious disease of cloven-hoofed animals that inflicts severe economic losses in the livestock industry. In 2009, FMDV serotype A caused outbreaks of FMD in cattle in China. Although an inactivated virus vaccine has proven effective to control FMD, its use may lead to new disease outbreaks due to a possible incomplete inactivation of the virus during the manufacturing process. Here, we expressed the P1-2A and the 3C coding regions of a serotype A FMDV field isolate in silkworm pupae (Bombyx mori and evaluated the immunogenicity of the expression products. Four of five cattle vaccinated with these proteins developed high titers of FMDV-specific antibody and were completely protected against virulent homologous virus challenge with 10,000 50% bovine infectious doses (BID(50. Furthermore, the 50% bovine protective dose (PD(50 test was performed to assess the bovine potency of the empty capsid subunit vaccine and was shown to achieve 4.33 PD(50 per dose. These data provide evidence that silkworm pupae can be used to express immunogenic FMDV proteins. This strategy might be used to develop a new generation of empty capsid subunit vaccines against a variety of diseases.

  16. Comparative Transcriptome Analysis of Bombyx mori (Lepidoptera) Larval Midgut Response to BmNPV in Susceptible and Near-Isogenic Resistant Strains

    Science.gov (United States)

    Geng, Lei; Xu, Jia-Ping; Yu, Dong; Zhang, Shang-Zhi; Ma, Yan; Fei, Dong-Qiong

    2016-01-01

    Bombyx mori nucleopolyhedrovirus (BmNPV) is one of the primary pathogens causing severe economic losses in sericulture. However, the molecular mechanism of silkworm resistance to BmNPV remains largely unknown. Here, the recurrent parent P50 (susceptible strain) and the near-isogenic line BC9 (resistance strain) were used in a comparative transcriptome study examining the response to infection with BmNPV. A total of 14,300 unigenes were obtained from two different resistant strains; of these, 869 differentially expressed genes (DEGs) were identified after comparing the four transcriptomes. Many DEGs associated with protein metabolism, cytoskeleton, and apoptosis may be involved in the host response to BmNPV infection. Moreover, some immunity related genes were also altered following BmNPV infection. Specifically, after removing genetic background and individual immune stress response genes, 22 genes were found to be potentially involved in repressing BmNPV infection. These genes were related to transport, virus replication, intracellular innate immune, and apoptosis. Our study provided an overview of the molecular mechanism of silkworm resistance to BmNPV infection and laid a foundation for controlling BmNPV in the future. PMID:27168061

  17. Comparative Transcriptome Analysis of Bombyx mori (Lepidoptera Larval Midgut Response to BmNPV in Susceptible and Near-Isogenic Resistant Strains.

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    Xue-Yang Wang

    Full Text Available Bombyx mori nucleopolyhedrovirus (BmNPV is one of the primary pathogens causing severe economic losses in sericulture. However, the molecular mechanism of silkworm resistance to BmNPV remains largely unknown. Here, the recurrent parent P50 (susceptible strain and the near-isogenic line BC9 (resistance strain were used in a comparative transcriptome study examining the response to infection with BmNPV. A total of 14,300 unigenes were obtained from two different resistant strains; of these, 869 differentially expressed genes (DEGs were identified after comparing the four transcriptomes. Many DEGs associated with protein metabolism, cytoskeleton, and apoptosis may be involved in the host response to BmNPV infection. Moreover, some immunity related genes were also altered following BmNPV infection. Specifically, after removing genetic background and individual immune stress response genes, 22 genes were found to be potentially involved in repressing BmNPV infection. These genes were related to transport, virus replication, intracellular innate immune, and apoptosis. Our study provided an overview of the molecular mechanism of silkworm resistance to BmNPV infection and laid a foundation for controlling BmNPV in the future.

  18. Silk fibroin film from golden-yellow Bombyx mori is a biocomposite that contains lutein and promotes axonal growth of primary neurons.

    Science.gov (United States)

    Pistone, Assunta; Sagnella, Anna; Chieco, Camilla; Bertazza, Gianpaolo; Varchi, Greta; Formaggio, Francesco; Posati, Tamara; Saracino, Emanuela; Caprini, Marco; Bonetti, Simone; Toffanin, Stefano; Di Virgilio, Nicola; Muccini, Michele; Rossi, Federica; Ruani, Giampiero; Zamboni, Roberto; Benfenati, Valentina

    2016-05-01

    The use of doped silk fibroin (SF) films and substrates from Bombyx mori cocoons for green nanotechnology and biomedical applications has been recently highlighted. Cocoons from coloured strains of B. mori, such as Golden-Yellow, contain high levels of pigments that could have a huge potential for the fabrication of SF based biomaterials targeted to photonics, optoelectronics and neuroregenerative medicine. However, the features of extracted and regenerated SF from cocoons of B. mori Golden-Yellow strain have never been reported. Here we provide a chemophysical characterization of regenerated silk fibroin (RSF) fibers, solution, and films obtained from cocoons of a Golden-Yellow strain of B. mori, by SEM, (1) H-NMR, HPLC, FT-IR, Raman and UV-Vis spectroscopy. We found that the extracted solution and films from B. mori Golden-Yellow fibroin displayed typical Raman spectroscopic and optical features of carotenoids. HPLC-analyses revealed that lutein was the carotenoid contained in the fiber and RSF biopolymer from yellow cocoons. Notably, primary neurons cultured on yellow SF displayed a threefold higher neurite length than those grown of white SF films. The results we report pave the way to expand the potential use of yellow SF in the field of neuroregenerative medicine and provide green chemistry approaches in biomedicine. © 2016 Wiley Periodicals, Inc. Biopolymers 105: 287-299, 2016. PMID:26756916

  19. Changes in growth and lipid profiles of silk gland, mid-gut biochemical composition of silkworm, Bombyx mori L. on exposure to prostaglandin F2alpha.

    Science.gov (United States)

    Miao, Yun-gen; Jiang, Li-jun

    2003-01-01

    The growth of the silkworm is influenced by the outside and inside environment. Among them, the category of various endocrine hormone of inside is the main factors that adjust the characters such as growth and propagate. In this experiment, we applied different dosage of prostaglandin to the fourth and fifth instar silkworm to observe the effects of prostaglandin F2alpha (PGF2alpha) on silk gland growth, mid-gut biochemical constituents and the lipid profiles of silkworm larva, Bombyx mori L. The weight of the posterior silk gland increased significantly (P mori L. indicate that PGF2alpha favored stimulatory effect on physiology of digestion, absorption and transportation of nutrients which might influence on the growth and development of larva. PMID:12611497

  20. Genetic analysis of the electrophysiological response to salicin, a bitter substance, in a polyphagous strain of the silkworm Bombyx mori.

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    Tetsuya Iizuka

    Full Text Available Sawa-J is a polyphagous silkworm (Bombyx mori L. strain that eats various plant leaves that normal silkworms do not. The feeding preference behavior of Sawa-J is controlled by one major recessive gene(s on the polyphagous (pph locus, and several minor genes; moreover, its deterrent cells possess low sensitivity to some bitter substances including salicin. To clarify whether taste sensitivity is controlled by the pph locus, we conducted a genetic analysis of the electrophysiological characteristics of the taste response using the polyphagous strain Sawa-J·lem, in which pph is linked to the visible larval marker lemon (lem on the third chromosome, and the normal strain Daiankyo, in which the wild-type gene of pph (+(pph is marked with Zebra (Ze. Maxillary taste neurons of the two strains had similar dose-response relationships for sucrose, inositol, and strychnine nitrate, but the deterrent cell of Sawa-J·lem showed a remarkably low sensitivity to salicin. The F(1 generation of the two strains had characteristics similar to the Daiankyo strain, consistent with the idea that pph is recessive. In the BF(1 progeny between F(1 females and Sawa-J·lem males where no crossing-over occurs, the lem and Ze phenotypes corresponded to different electrophysiological reactions to 25 mM salicin, indicating that the gene responsible for taste sensitivity to salicin is located on the same chromosome as the lem and Ze genes. The normal and weak reactions to 25 mM salicin were segregated in crossover-type larvae of the BF(1 progeny produced by a reciprocal cross, and the recombination frequency agreed well with the theoretical ratio for the loci of lem, pph, and Ze on the standard linkage map. These results indicate that taste sensitivity to salicin is controlled by the gene(s on the pph locus.

  1. Genetic analysis of the electrophysiological response to salicin, a bitter substance, in a polyphagous strain of the silkworm Bombyx mori.

    Science.gov (United States)

    Iizuka, Tetsuya; Tamura, Toshiki; Sezutsu, Hideki; Mase, Keisuke; Okada, Eiji; Asaoka, Kiyoshi

    2012-01-01

    Sawa-J is a polyphagous silkworm (Bombyx mori L.) strain that eats various plant leaves that normal silkworms do not. The feeding preference behavior of Sawa-J is controlled by one major recessive gene(s) on the polyphagous (pph) locus, and several minor genes; moreover, its deterrent cells possess low sensitivity to some bitter substances including salicin. To clarify whether taste sensitivity is controlled by the pph locus, we conducted a genetic analysis of the electrophysiological characteristics of the taste response using the polyphagous strain Sawa-J·lem, in which pph is linked to the visible larval marker lemon (lem) on the third chromosome, and the normal strain Daiankyo, in which the wild-type gene of pph (+(pph)) is marked with Zebra (Ze). Maxillary taste neurons of the two strains had similar dose-response relationships for sucrose, inositol, and strychnine nitrate, but the deterrent cell of Sawa-J·lem showed a remarkably low sensitivity to salicin. The F(1) generation of the two strains had characteristics similar to the Daiankyo strain, consistent with the idea that pph is recessive. In the BF(1) progeny between F(1) females and Sawa-J·lem males where no crossing-over occurs, the lem and Ze phenotypes corresponded to different electrophysiological reactions to 25 mM salicin, indicating that the gene responsible for taste sensitivity to salicin is located on the same chromosome as the lem and Ze genes. The normal and weak reactions to 25 mM salicin were segregated in crossover-type larvae of the BF(1) progeny produced by a reciprocal cross, and the recombination frequency agreed well with the theoretical ratio for the loci of lem, pph, and Ze on the standard linkage map. These results indicate that taste sensitivity to salicin is controlled by the gene(s) on the pph locus. PMID:22649537

  2. Scientific validation of cardioprotective attribute by standardized extract of Bombyx mori against doxorubicin-induced cardiotoxicity in murine model.

    Science.gov (United States)

    Khan, Masood S; Singh, Mhaveer; Khan, Mohammad A; Arya, D S; Ahmad, Sayeed

    2014-01-01

    Doxorubicin (DOX) is an excellent antineoplastic agent used for the treatment of hematological and solid malignancies. The aqueous extract of Bombyx mori (BMAE) contains amino acids and some flavonoids with obvious cardioprotective effect. The aim of this study was to investigate the possible protective effect of BMAE against DOX-induced cardiotoxicity and its underlying mechanisms on murine model. The metabolic profiling of BMAE was carried out by Ultra Performance Liquid Chromatography-Mass Spectrometry (UPLC-MS) and the amino acid profiling by HPLC method using fluorescence detector (HPLC-FLD). The biochemical parameter like caspase-3, tumor necrosis factor-alpha (TNF-α), interleukin -6 (IL-6), creatine kinase-MB (CK-MB), lactate dehydrogenase (LDH) and malondialdehyde (MDA) were studied. Tissue damage was further evaluated by histopathological studies. The metabolic profiling of BMAE exhibited presence of quercetin 7-O-ß-D-glucoside, kaempferol 7-O-ß-D-glucopyranoside, coumaric acid glucoside, 2-hydroxy-nonadecanoic acid and 9,12-dihydroxy stearic acid as important constituents. The amino acid profile by HPLC-FLD showed presence of 17 amino acids. The BMAE showed prominent free radical scavenging activity when assessed by the H2O2 and super-oxide method. The results of present investigation showed protection against DOX-induced oxidative stress (lipid peroxidation), by reverting activities of apoptotic markers (caspase-3 and TNF-α), cardiac markers (CK-MB and LDH activities) as well as pro-inflammatory marker IL-6 followed by oral administration of BMAE. In addition, results of histopathology also supported well the above results. It was observed that BMAE protects DOX-induced cardiotoxicity by virtue of its antioxidants possibly by flavonoids and amino acids. PMID:26417320

  3. Fibroin and sericin from Bombyx mori silk stimulate cell migration through upregulation and phosphorylation of c-Jun.

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    Celia Martínez-Mora

    Full Text Available Wound healing is a biological process directed to the restoration of tissue that has suffered an injury. An important phase of wound healing is the generation of a basal epithelium able to wholly replace the epidermis of the wound. A broad range of products derived from fibroin and sericin from Bombyx mori silk are used to stimulate wound healing. However, so far the molecular mechanism underlying this phenomenon has not been elucidated. The aim of this work was to determine the molecular basis underlying wound healing properties of silk proteins using a cell model. For this purpose, we assayed fibroin and sericin in a wound healing scratch assay using MDA-MB-231 and Mv1Lu cells. Both proteins stimulated cell migration. Furthermore, treatment with sericin and fibroin involved key factors of the wound healing process such as upregulation of c-Jun and c-Jun protein phosphorylation. Moreover, fibroin and sericin stimulated the phosphorylation of ERK 1/2 and JNK 1/2 kinases. All these experiments were done in the presence of specific inhibitors for some of the cell signalling pathways referred above. The obtained results revealed that MEK, JNK and PI3K pathways are involved in fibroin and sericin stimulated cells migration. Inhibition of these three kinases prevented c-Jun upregulation and phosphorylation by fibroin or sericin. Fibroin and sericin were tested in the human keratinocyte cell line, HaCaT, with similar results. Altogether, our results showed that fibroin and sericin initiate cell migration by activating the MEK, JNK and PI3K signalling pathways ending in c-Jun activation.

  4. Cloning and Expression Profile of Deoxyhypusine Snyhtase Gene and Deoxyhypusine Hydroxylase Gene in Silkworm,Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    WANG Geng-xian; SIMA Yang-hu; ZHANG Sheng-xiang; XU Shi-qing

    2009-01-01

    Deoxyhypusine snyhtase (DHS) and deoxyhypusine hydroxylase (DOHH) are the two enzymes that catalyze the synthesis of hypusine within eukaryotic initiation factor 5A (eIF5A).Synthesis of hypusine is essential for the function of eIF5A in eukaryotic cell proliferation and survival.Here we described the cloning and expression of two full-length cDNAs,encoding respectively DHS-like protein and DOHH-like protein from Bombyx mori by using the methods of bioinformatics,RACE,and RT-PCR technology,named as BmDHS and BmDOHH.Sequencing results indicate that they are 1311 and 1874 bp in length including complete open reading frame (ORF) 1116 and 915 bp,which encode 371 amino acids (molecular weight is about 41.11 kD and isoelectric point is 5.84) and 304 amino acids (molecular weight is about 34.30 kD and isoelectric point is 4.86),respectively.BmDHS contains only 1 exon,and BmDOHH contains 4 exons and 3 introns.The deduced amino acid sequence of BmDHS contains a deoxyhypusine synthase domain from 47 to 361 amino acid residues,and the deduced amino acid sequence of BmDOHH contains 6 E-Z type HEAT repeat domains (23-52,54-83,87-116,177-206,208-237,and 241-270).Compared to DHS and DOHH amino acid sequences from other species,such as Homo sapiens and Drosophila melanogaster,both silkworm DHS protein and DOHH protein have more than 55% identity.The conservative regions are very similar with each other.The phylogenetic tree analysis indicated that not only DHS but also DOHH from different species has genus-specific features.The expressions of BmDHS and BmDOHH are no tissue and stage specific in our tested samples.

  5. Specific expression of GFPuv-β1,3-N-acetylglucosaminyltransferase 2 fusion protein in fat body of Bombyx mori silkworm larvae using signal peptide

    International Nuclear Information System (INIS)

    Bombyxin (bx) and prophenoloxidase-activating enzyme (ppae) signal peptides from Bombyx mori, their modified signal peptides, and synthetic signal peptides were investigated for the secretion of GFPuv-β1,3-N-acetylglucosaminyltransferase 2 (GGT2) fusion protein in B. mori Bm5 cells and silkworm larvae using cysteine protease deficient B. mori multiple nucleopolyhedrovirus (BmMNPV-CP- ) and its bacmid. The secretion efficiencies of all signal peptides were 15-30% in Bm5 cells and 24-30% in silkworm larvae, while that of the +16 signal peptide was 0% in Bm5 cells and 1% in silkworm larvae. The fusion protein that contained the +16 signal peptide was expressed specifically in the endoplasmic reticulum (ER) and in the fractions of cell precipitations. Ninety-four percent of total intracellular β1,3-N-acetylglucosaminyltransferase (β3GnT) activity was detected in cell precipitations following the 600, 8000, and 114,000g centrifugations. In the case of the +38 signal peptide, 60% of total intracellular activity was detected in the supernatant from the 114,000g spin, and only 1% was found in the precipitate. Our results suggest that the +16 signal peptide might be situated in the transmembrane region and not cleaved by signal peptidase in silkworm or B. mori cells. Therefore, the fusion protein connected to the +16 signal peptide stayed in the fat body of silkworm larvae with biological function, and was not secreted extracellularly

  6. 20-Hydroxyecdysone stimulates nuclear accumulation of BmNep1, a nuclear ribosome biogenesis-related protein in the silkworm, Bombyx mori.

    Science.gov (United States)

    Ji, M-M; Liu, A-Q; Sima, Y-H; Xu, S-Q

    2016-10-01

    The pathway of communication between endocrine hormones and ribosome biogenesis critical for physiological adaptation is largely unknown. Nucleolar essential protein 1 (Nep1) is an essential gene for ribosome biogenesis and is functionally conserved in many in vertebrate and invertebrate species. In this study, we cloned Bombyx mori Nep1 (BmNep1) due to its high expression in silk glands of silkworms on day 3 of the fifth instar. We found that BmNep1 mRNA and protein levels were upregulated in silk glands during fourth-instar ecdysis and larval-pupal metamorphosis. By immunoprecipitation with the anti-BmNep1 antibody and liquid chromatography-tandem mass spectrometry analyses, it was shown that BmNep1 probably interacts with proteins related to ribosome structure formation. Immunohistochemistry, biochemical fractionation and immunocytochemistry revealed that BmNep1 is localized to the nuclei in Bombyx cells. Using BmN cells originally derived from ovaries, we demonstrated that 20-hydroxyecdysone (20E) induced BmNep1 expression and stimulated nuclear accumulation of BmNep1. Under physiological conditions, BmNep1 was also upregulated in ovaries during larval-pupal metamorphosis. Overall, our results indicate that the endocrine hormone 20E facilitates nuclear accumulation of BmNep1, which is involved in nuclear ribosome biogenesis in Bombyx. PMID:27329527

  7. Parental environmental exposure leads to glycometabolic disturbances that affect fertilization of eggs in the silkworm Bombyx mori: the parental transcript legacy.

    Science.gov (United States)

    Tao, H; Liu, H J; Cheng, Y Q; Sima, Y H; Yin, W M; Xu, S Q

    2015-01-01

    Parental transcript legacy plays an important role in fertilization and development of the early embryo. Parental environmental exposure affects the fertilization of eggs, but the underlying biochemical mechanism is largely unresolved. In this study, the parental environmental effects on fertilization of eggs were explored in the silkworm Bombyx mori (B. mori), an ideal lepidopteran animal model. The results showed that the rate of fertilization decreased after the parents were exposed to a poor environment at 32 °C with continuous illumination for 72 h on days 6-9 of the pupal stage, which is a key period for germ cell maturation. This was likely attributable to lower energy charge values, obstructed nicotinamide adenine dinucleotide (NAD(+)) regeneration and inactive tricarboxylic acid cycle (TCA), leading to accumulation of large amounts of pyruvic acid and lactic acid. This effect was related to energy metabolism via glycolysis; in particular disruption of pyruvate metabolism. In conclusion, this study showed parental exposure to an abnormal environment during germ cell maturation affected glycolysis and the subsequent fertilization of eggs via the parental transcript legacy in B. mori. PMID:25240789

  8. A cadherin-like protein functions as a receptor for Bacillus thuringiensis Cry1Aa and Cry1Ac toxins on midgut epithelial cells of Bombyx mori larvae.

    Science.gov (United States)

    Hara, Hirotaka; Atsumi, Shogo; Yaoi, Katsuro; Nakanishi, Kazuko; Higurashi, Satoshi; Miura, Nami; Tabunoki, Hiroko; Sato, Ryoichi

    2003-03-13

    Aminopeptidase N (APN) and cadherin-like protein (BtR175) from Bombyx mori larvae were examined for their roles in Cry1Aa- and Cry1Ac-induced lysis of B. mori midgut epithelial cells (MECs). APNs and BtR175 were present in all areas of the midgut, were particularly abundant in the posterior region, and were found only on columnar cell microvilli and not on the lateral membrane that makes cell-cell contacts. This distribution was in accordance with the distribution of Cry1A-susceptible MECs in the midgut. The lytic activity of Cry1Aa and Cry1Ac on collagenase-dissociated MECs was linearly dependent on toxin concentration. Although pre-treatment of MECs with anti-BtR175 antibody was observed to partially inhibit the lytic activity exerted by 0.1-1 nM Cry1Aa toxin or 5 nM Cry1Ac toxin, no significant inhibition was observed when MECs were pre-treated with anti-APN antibody. These results suggest that BtR175 functions as a major receptor for Cry1A toxins in the midgut of B. mori larvae. PMID:12633848

  9. Molecular cloning of a functional allatostatin gut/brain receptor and an allatostatin preprohormone from the silkworm Bombyx mori

    DEFF Research Database (Denmark)

    Secher, Thomas; Lenz, C; Cazzamali, G;

    2001-01-01

    on the addition of 4 x 10(-9)M Bombyx A-type allatostatins with a second messenger cascade (measured as bioluminescence), showing that BAR is a functional A-type allatostatin receptor. Southern blots suggest that Bombyx has at least one other BAR-related gene in addition to the BAR gene described in this paper...

  10. Identification and Characterization of Bacillus cereus SW7-1 in Bombyx mori (Lepidoptera: Bombycidae)

    OpenAIRE

    Li, Guan-Nan; Xia, Xue-Juan; Zhao, Huan-Huan; Sendegeya, Parfait; Zhu, Yong

    2015-01-01

    The bacterial diseases of silkworms cause significant reductions in sericulture and result in huge economic loss. This study aimed to identify and characterize a pathogen from diseased silkworm. SW7-1, a pathogenic bacterial strain, was isolated from the diseased silkworm. The strain was identified on the basis of its bacteriological properties and 16S rRNA gene sequence. The colony was round, slightly convex, opaque, dry, and milky on a nutrient agar medium, the colony also exhibited jagged ...

  11. Proteomic-based insight into Malpighian tubules of silkworm Bombyx mori.

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    Xiao-wu Zhong

    Full Text Available Malpighian tubules (MTs are highly specific organs of arthropods (Insecta, Myriapoda and Arachnida for excretion and osmoregulation. In order to highlight the important genes and pathways involved in multi-functions of MTs, we performed a systematic proteomic analysis of silkworm MTs in the present work. Totally, 1,367 proteins were identified by one-dimensional gel electrophoresis coupled with liquid chromatography-tandem mass spectrometry, and as well as by Trans Proteomic Pipeline (TPP and Absolute protein expression (APEX analyses. Forty-one proteins were further identified by two-dimensional gel electrophoresis. Some proteins were revealed to be significantly associated with various metabolic processes, organic solute transport, detoxification and innate immunity. Our results might lay a good foundation for future functional studies of MTs in silkworm and other lepidoptera.

  12. Genome fingerprinting of the silkworm, Bombyx mori, using random arbitrary primers.

    Science.gov (United States)

    Nagaraja, G M; Nagaraju, J

    1995-09-01

    The random amplified polymorphic DNA (RAPD) technique was used to study DNA profiling of thirteen silkworm genotypes. The genotypes included six diapausing and seven nondiapausing varieties that represent a high degree of divergence with respect to geographic origin, and morphological, qualitative, quantitative and biochemical characters. Two hundred sixteen amplified products were generated using 40 random primers. Genotype-specific amplification products were identified. Amplification products specific to diapausing genotypes were also identified. Segregation of the RAPD marker was analyzed in a backcross population and found to be inherited as dominant Mendelian traits. Based on pairwise comparison of amplified products, the genetic similarity was performed by a hierarchical clustering technique. Silkworm genotypes were clustered into two groups, one consisting of six diapausing and the other of seven nondiapausing genotypes. The results of our study suggest that the RAPD technique could be used as a powerful tool to generate genetic markers that are linked to traits of interest in the silkworm. PMID:8582347

  13. CYP18A1 regulates tissue-specific steroid hormone inactivation in Bombyx mori

    OpenAIRE

    Li, Zhiqian; Ge, Xie; Ling, Lin; Zeng, Baosheng; Xu, Jun; Aslam, Abu F.M.; You, Lang; Palli, Subba Reddy; Huang, Yongping; Tan, Anjiang

    2014-01-01

    Insect development and metamorphosis are regulated by two major hormones, juvenile hormone and ecdysteroids. Despite being the key regulator of insect developmental transitions, the metabolic pathway of the primary steroid hormone, 20-hydroxyecdysone (20E), especially its inactivation pathway, is still not completely elucidated. A cytochrome P450 enzyme, CYP18A1, has been shown to play key roles in insect steroid hormone inactivation through 26-hydroxylation. Here, we identified two CYP18 (Bm...

  14. Identification and Characterization of Bacillus cereus SW7-1 in Bombyx mori (Lepidoptera: Bombycidae).

    Science.gov (United States)

    Li, Guan-Nan; Xia, Xue-Juan; Zhao, Huan-Huan; Sendegeya, Parfait; Zhu, Yong

    2015-01-01

    The bacterial diseases of silkworms cause significant reductions in sericulture and result in huge economic loss. This study aimed to identify and characterize a pathogen from diseased silkworm. SW7-1, a pathogenic bacterial strain, was isolated from the diseased silkworm. The strain was identified on the basis of its bacteriological properties and 16S rRNA gene sequence. The colony was round, slightly convex, opaque, dry, and milky on a nutrient agar medium, the colony also exhibited jagged edges. SW7-1 was Gram-positive, without parasporal crystal, and 0.8-1.2 by 2.6-3.4 µm in length, resembling long rods with rounded ends. The strain was positive to most of the physiological biochemical tests used in this study. The strain could utilize glucose, sucrose, and maltose. The results of its 16S rRNA gene sequence analysis revealed that SW7-1 shared the highest sequence identity (>99%) with Bacillus cereus strain 14. The bacterial strain was highly susceptible to gentamycin, streptomycin, erythromycin, norfloxacin, and ofloxacin and moderately susceptible to tetracycline and rifampicin. It exhibited resistance to other antibiotics. SW7-1 had hemolytic activity and could produce extracellular casease, lipase, and amylase. SW7-1 could reproduce septicemia-like symptoms with high mortality rate when re-fed to healthy silkworm. .The median lethal concentration (LC50) was 5.45 × 10(4) cfu/ml. Thus, SW7-1 was identified as B. cereus, which is a pathogen for silkworm and human infections are possible. PMID:26411789

  15. Expression of the fructose receptor BmGr9 and its involvement in the promotion of feeding, suggested by its co-expression with neuropeptide F1 in Bombyx mori.

    Science.gov (United States)

    Mang, Dingze; Shu, Min; Tanaka, Shiho; Nagata, Shinji; Takada, Tomoyuki; Endo, Haruka; Kikuta, Shingo; Tabunoki, Hiroko; Iwabuchi, Kikuo; Sato, Ryoichi

    2016-08-01

    Insect gustatory receptors (Grs) are members of a large family of proteins with seven transmembrane domains that provide insects with the ability to detect chemical signals critical for feeding, mating, and oviposition. To date, 69 Bombyx mori Grs (BmGrs) genes have been identified via genome studies. BmGr9 has been shown to respond specifically to fructose and to function as a ligand-gated ion channel selectively activated by fructose. However, the sites where this Gr are expressed remain unclear. We demonstrated using reverse transcription (RT)-PCR that BmGr9 is widely expressed in the central nervous system (CNS), as well as oral sensory organs. Additionally, immunohistochemistry was performed using anti-BmGr9 antiserum to show that BmGr9 is expressed in cells of the oral sensory organs, including the maxillary galea, maxillary palps, labrum, and labium, as well as in putative neurosecretory cells of the CNS. Furthermore, double immunohistochemical analysis showed that most BmGr9-expressing cells co-localized with putative neuropeptide F1-expressing cells in the brain, suggesting that BmGr9 is involved in the promotion of feeding behaviors. In addition, a portion of BmGr9-expressing cells in the brain co-localized with cells expressing BmGr6, a molecule of the sugar receptor clade, suggesting that sugars other than fructose are involved in the regulation of feeding behaviors in B. mori larvae. PMID:27288056

  16. Expression of human endostatin in larvae of silkworm (Bombyx mori) and in vitro activity assays.

    Science.gov (United States)

    Yongfeng, Jin; Yingfei, Wang; Zhenhong, Zhu; Yaozhou, Zhang

    2002-08-01

    Human endostatin is a novel antiangiogenic molecule, which can inhibit the proliferation and development of new blood vessels, and experimentally can cause nearly complete regression of established tumors. In this paper, the cDNA encoding human endostatin was cloned into a baculovirus shuttle vector pBacPAK8 and co-infected with linearized Bm-BacPAK6 DNA into and BmN cells. The recombinant virus was screened and identified by PCR, DNA and RNA dot hybridization, and ELISA assay. The recombinant endostatin was expressed in culture cells, and the larvae and pupa of silkworm by inoculation of recombinant virus. The biological activity assay showed that the expression product in larvae was over 150 microg/ml, about 50-fold higher than that expressed in cultured cells. SDS-PAGE and Western blotting analysis showed a pattern of molecular weight of about 20 kDa. The bio-activity of the protein product was determined by human umbilical vein endothelial cells (ECV304) proliferation test in vitro and the chick chorioallantoic membrane (CAM) vascular inhibition test. Endostatin showed significant inhibitory effect on endothelial cells in a dose-dependent manner. Silkworm-produced endostatin induced apoptosis of endothelial cells and also inhibited angiogenesis in the CAM assay. Combination regimen using angiostatin and endostatin showed more than additive effect in angiogenic inhibition and increasing apoptosis when compared with treatment with the individual antiangiogenic protein. PMID:12186748

  17. Backcross Breeding and Directional Selection of Two Multivoltines, N+p and Np of Silkworm, Bombyx mori L. for Higher Viability and Productivity in Eastern India

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    T. K. Mukhopadhyay

    2013-08-01

    Full Text Available A new superior multivoltine breed of Bombyx mori L. (D+p has been developed by crossing between two strains of Nistari Plain (Np and Nistari Marked (N+p which are the two most preferred strains of multivoltine breed at the farmer’s level of eastern India. Through standard breeding plan this needbased new breed, D+p was developed by adopting three steps: i Development of Recurrent Backcross Line through backcross breeding for initial eight generations; ii Improvement of developed RBL following directional selection process for the next twenty generations and ultimately iii stabilization for next eight generations upto F40 generation at normal environmental conditions. Study of nine quantitative characters related to viability and productivity of the new breed in relation to its parents have been studied statistically and Heterosis%.

  18. Analysis of four achaete-scute homologs in Bombyx mori reveals new viewpoints of the evolution and functions of this gene family

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    Yi Yongzhu

    2008-03-01

    Full Text Available Abstract Background achaete-scute complexe (AS-C has been widely studied at genetic, developmental and evolutional levels. Genes of this family encode proteins containing a highly conserved bHLH domain, which take part in the regulation of the development of central nervous system and peripheral nervous system. Many AS-C homologs have been isolated from various vertebrates and invertebrates. Also, AS-C genes are duplicated during the evolution of Diptera. Functions besides neural development controlling have also been found in Drosophila AS-C genes. Results We cloned four achaete-scute homologs (ASH from the lepidopteran model organism Bombyx mori, including three proneural genes and one neural precursor gene. Proteins encoded by them contained the characteristic bHLH domain and the three proneural ones were also found to have the C-terminal conserved motif. These genes regulated promoter activity through the Class A E-boxes in vitro. Though both Bm-ASH and Drosophila AS-C have four members, they are not in one by one corresponding relationships. Results of RT-PCR and real-time PCR showed that Bm-ASH genes were expressed in different larval tissues, and had well-regulated expressional profiles during the development of embryo and wing/wing disc. Conclusion There are four achaete-scute homologs in Bombyx mori, the second insect having four AS-C genes so far, and these genes have multiple functions in silkworm life cycle. AS-C gene duplication in insects occurs after or parallel to, but not before the taxonomic order formation during evolution.

  19. Esterase isozyme polymorphism, specific and nonspecific esterase, syngenic lines development and natural occurrence of a thermostable esterase in the tropical silkworm Bombyx mori L.

    Science.gov (United States)

    Chattopadhyay, G K; Sengupta, A K; Verma, A K; Sen, S K; Saratchandra, B

    2001-11-01

    Esterase isozyme polymorphism was documented for digestive juice and haemolymph of the tropical multivoltine silkworm, Bombyx mori L., breed CB5 (GP) and its syngenic lines (CB5Lm(e)-1, CB5Lm-2 and CB5Lm-5) using alpha- and beta-naphthylacetate separately as nonspecific substrates (Ogita, Z., Kasai, T., 1965. Genetico-biochemical analysis of specific esterases in Musca domestica. Jpn. J. Genet. 40, 173-184). Polymorphism existed in the isozyme pattern of alpha-esterase with two or three bands in digestive juice and three to five bands in haemolymph. No polymorphism was observed in beta-esterase isozyme pattern having four bands in digestive juice and two bands in haemolymph. During the course of esterase isozyme studies, the presence of some specific alpha-esterase bands (Est-1, 4 and 5) in haemolymph and beta-esterase bands (Est-1, 2 and 3) in digestive juice were observed. But both alpha- and beta-esterase bands Est-3 and 4 in digestive juice and Est-2 and 3 in haemolymph were found to be nonspecific. Nonspecific beta-esterase band (Est-3) in haemolymph of CB5 (GP) and its syngenic lines withstood a temperature up to 80+/-1 degrees C for 10 min. No thermostable band was observed in the isozyme zymogram of alpha-esterase in digestive juice and haemolymph or beta-esterase in digestive juice. Overall, this study discusses the presence of esterase heterogeneity in the CB5 (GP) genepool, syngenic lines development, occurrence of specific alpha- and beta-esterase bands in digestive juice and haemolymph and thermostable beta-esterase band Est-3 in haemolymph in tropical silkworm Bombyx mori L. PMID:11583932

  20. Analysis of ESTs and gene expression patterns of the posterior silkgland in the fifth instar larvae of silkworm, Bombyx mori L.

    Institute of Scientific and Technical Information of China (English)

    ZHONG Boxiong; LOU Chengfu; YU Yingpo; XU Yusong; YU Hong; LU Xingmeng; MIAO Yungen; YANG Jun; XU Hao; HU Songnian

    2005-01-01

    The fibroin gene expression pattern and regulation of the posterior silkgland were studied by means of expressed sequence tags (ESTs) using the first and fifth day larvae of the fifth instar of silkworm, Bombyx mori L (strain: C 108). The results showed that there were 911 repetitive ESTs and 1950 single sequences (Singlets) among total 2861 consentient sequences, which were spliced. 1335 sequences were identified and the other 1526 were unknown. 5560 sequences (55.89%) in the posterior silkgland cell of the silkworm were new ESTs without homology with EST data published by Mita et al. The number of repetitive ESTs and single sequences from the first day larvae of the fifth instar was double more than that of the fifth day of the same instar in the silkworms. The unigenes which were more than 50 in repetitive EST size (contig size) came to only about 0.5% in total consentient sequences. There were significant differences between gene expression frequencies, and expressed genes were related to fibroin synthesis and its secretion and fibroin composition. Comparing the fifth day with the first day of the fifth instar, the genes-expressed quantity of fibroin heavy-chain gene was 18 fold higher, fibroin light-chain gene 9 fold and fibroin P52 gene 8 fold. 508 genes functioned for cellular component and 315 for enzyme after function tracing. These results implied that the gene expression of the first day was mainly for preparation for fibroin synthesis except for the growth of silkgland cells, and the gene expression of the fifth day of the fifth instar was mainly for synthesizing and excreting fibroin. Because the ratio of heavy chain, light chain and p25 of fibroin was not 6:6:1 as theoretically expected, or its special H-chain structure, the H-chain gene was not easy to detect through EST technique. Most of genes among total 2861 consentient sequences functioned for fibroin synthesis and secretion. This suggested the fibroin synthesis and secretion procedure of the

  1. Influência de genótipos de amoreira (Morus sp. e substratos no peso e características de casulos do bicho-da-seda (Bombyx mori L. Influence of mulberry (Morus sp. genotypes and substrates in weight and characteristics of silkworm cocoons (Bombyx mori L.

    Directory of Open Access Journals (Sweden)

    Odinete Murari

    2001-05-01

    Full Text Available Avaliou-se o efeito de três genótipos de amoreira, Morus sp. (Moraceae: Miura, FM Shima Miura e IZ 56/4, três tipos de esteiras de criação: terra compactada, concreto e tela plástica sobre o peso e algumas características industriais de casulos produzidos pelo bicho-da-seda, Bombyx mori (Lepidoptera. Houve influência das interações de genótipos com esteiras de criação sobre o peso dos casulos produzidos. Com relação às características industriais, os tratamentos que mais se sobressaíram foram: Miura / terra compactada, FM-SM / tela plástica, IZ 56/4 / terra compactada e IZ 56/4 / tela plástica.The effect of three genotypes of mulberry, Morus sp. (Moraceae, namely, Miura, FM Shima Miura and IZ 56/4, and three types of rearing substrate comprising compact soil, concrete and plastic screen were estimated on weight and on certain industrial characteristics of Bombyx mori (Lepidoptera cocoons. Genotype interactions with rearing substrates affected weight of cocoons produced. Analysis showed the best treatments for manufacturer parameters were Miura / compact soil, FM-Shima Miura / plastic screen, IZ 56/4 / compact soil and IZ 56/4 / plastic screen.

  2. Progeny of Palmistichus elaeisis Delvare and LaSalle (Hymenoptera: Eulophidae) parasitising pupae of Bombyx mori L. (Lepidoptera: Bombycidae) of different ages

    International Nuclear Information System (INIS)

    Palmistichus elaeisis Delvare and LaSalle is a natural pupal parasitoid of eucalyptus defoliator lepidopterans and is considered a promising biocontrol agent. However, the development of efficient rearing techniques for this natural enemy are fi rst required before it can be used in biocontrol programs. Bombyx mori L. pupae are potential alternative hosts for this parasitoid mass rearing, and they are easy to rear. Therefore, we investigated the most suitable host age and the effects of parasitoid age on progeny production of P. elaeisis. B. mori pupae, 24 h-, 48 h-, 72 h- or 96 h-old were exposed to P. elaeisis females of similar age. The duration of the life cycle (egg-adult) of P. elaeisis was not affected by the age of the parasitizing female; however, the host age affected parasitoid development. The best parasitization was obtained for 72h- to 96h-old parasitoid females when offered to 48h- to 72h-old host pupae, allowing the synchronized rearing of a large number of P. elaeisis offspring. (author)

  3. Demarcation of diapause development by cold and its relation to time-interval activation of TIME-ATPase in eggs of the silkworm, Bombyx mori.

    Science.gov (United States)

    Ti, Xiaonan; Tuzuki, Nobuhiko; Tani, Naoki; Morigami, Etsuko; Isobe, Minoru; Kai, Hidenori

    2004-11-01

    We investigated the mode of action of winter cold in the termination of diapause by investigating Time-Interval-Measuring Enzyme (TIME). First, we determined the period of cold required for the completion of diapause development. Synchronously developing egg batches of a pure strain (C108 Bombyx mori silkworm) were used to minimize variations in hatching time. Hatching occurred with only 18 days chilling at 5 degrees C when the incubation at 25 degrees C after the chilling was elongated. The 18-day period was much shorter than we expected; diapause in B. mori is known to terminate completely with about 100 days of chilling. Even in such a short period of chilling, no sporadic hatching occurred. Moreover, we determined that a temperature-insensitive stage, which we called "Neboke", followed the short cold-requiring stage. Thus, the stage of diapause development was demarcated from other stages of diapause. While the length of diapause development was elongated when chilling was delayed after oviposition, the Neboke stage length was invariant. Cold evidently exerts its effect only on diapause development. When TIME was purified from eggs and chilled in test tubes, a transitory burst of its ATPase activity occurred at a time equivalent to shortly before the completion of diapause development; this was an interval-timer activation. The mechanism by which cold activates TIME to measure the time interval may help explain in biochemical terms the insect's adaptation to its seasonal environments. PMID:15607508

  4. Aminopeptidase N isoforms from the midgut of Bombyx mori and Plutella xylostella -- their classification and the factors that determine their binding specificity to Bacillus thuringiensis Cry1A toxin.

    Science.gov (United States)

    Nakanishi, Kazuko; Yaoi, Katsuro; Nagino, Yasushi; Hara, Hirotaka; Kitami, Madoka; Atsumi, Shogo; Miura, Nami; Sato, Ryoichi

    2002-05-22

    Novel aminopeptidase N (APN) isoform cDNAs, BmAPN3 and PxAPN3, from the midguts of Bombyx mori and Plutella xylostella, respectively, were cloned, and a total of eight APN isoforms cloned from B. mori and P. xylostella were classified into four classes. Bacillus thuringiensis Cry1Aa and Cry1Ab toxins were found to bind to specific APN isoforms from the midguts of B. mori and P. xylostella, and binding occurred with fragments that corresponded to the BmAPN1 Cry1Aa toxin-binding region of each APN isoform. The results suggest that APN isoforms have a common toxin-binding region, and that the apparent specificity of Cry1Aa toxin binding to each intact APN isoform seen in SDS-PAGE is determined by factors such as expression level in conjunction with differences in binding affinity. PMID:12023048

  5. Effect of X-ray irradiation on the male moths of two voltine groups of the silkworm, Bombyx mori, and inheritance of induced sterility

    International Nuclear Information System (INIS)

    Full text: The silkworm, Bombyx mori, one of the important sericigenous insect members, during its life cycle from egg to adult exhibits sequential events of spermatogenesis in homogametic males and mature sperms are invariably seen in adult moths. In the present experiment, the male moths of two races of Bombyx mori viz., Pure Mysore and Kalimpong-A belonging to two voltine groups namely multivoltine and bivoltines, respectively were irradiated with X-rays at two independent doses of 50 Gy and 100 Gy and were crossed with unmated female moths. Untreated batches were maintained as control. The eggs deposited by bivoltine race were treated with hydrochloric acid to break the diapause and were allowed to develop at 25±1 deg. C and 80% RH. Unfertilised and unhatched eggs of both the parental races were examined by adopting standard procedures and the hatched larvae were reared under standard rearing conditions. In order to understand the inherited sterility, the untreated females were crossed with F1, F2 and F3 males survived from X-ray treatment. The results have clearly indicated that, in the parental generation, unfertilised and unhatched eggs ranged from 65-75% with the multivoltine race revealing fewer of embryonic deaths and unfertilised eggs and in both races a dose dependency was found. The eggs laid by F1 moths recorded the highest number of unfertilized and unhatched eggs while in F2 and F3 populations there was a gradual decline in the sterility resulting in a significant increase in egg hatch. The data obtained were statistically analysed at every generation and compared with those of untreated control batches. The high percentage of sterility in the treated F1 and inherited sterility in F2 and F3 populations are discussed in the light of chromosome imbalance (translocation) that may be able to pass through meiotic division in adult moths, which may cause production of functionless sperms resulting in delayed lethal effect from irradiation. The

  6. Identification and Expression Patterns of Atlastin Genes in Silkworm, Bombyx mori%家蚕Atlastin基因(BmATL)的鉴定及表达模式

    Institute of Scientific and Technical Information of China (English)

    陈全梅; 谭祥; 杨强; 胡晓明; 马振刚; 赵萍

    2011-01-01

    The hereditary spastic paraplegias (HSPs) are a group of inherited human neurological disorders causing increased stiffness and overactive muscle reflexes. Among genes underlying HSPs, Atlastin encodes a protein with a typicai GTP binding (GBP) domain and two adjacent transmembrane regions. Atlastin is located on membranes of Golgi apparatus and endoplasmic reticulum, involving in vesicle trafficking. By searching against the silkworm (Bombyx mori)genome sequence, we identified 4 Atlastin genes, designated as BmATL1 to BmATL4, on the basis of homology analysis with Atlastin genes from Homo sapiens and Drosophila melanogaster. All four BmATLs have a conserved dynamin-like GBP domain, among which BmATL1 and BmATL2 have a molecular mass of about 60 kD with two adjacent transmembrane regions, and BmATL3 and BmATL4 have a molecular mass of about 85 kD without transmembrane region. Expression pattern analyses revealed that BmATL1 and BmATL2 were lowly expressed in all tissues of silkworm larvae on day 3 of the 5th instar, while BmATL3 and BmATL4 were highly expressed in silkworm hemocytes. The above analyses suggest that BmATL1 is similar to Atlastin of H. sapiens and D. melanogaster in molecular mass, structural domains, and expression pattern.%Atlastin基因是人类遗传性痉挛性截瘫(hereditary spastic paraplegia,HSP)疾病的致病基因之一.Atlastin蛋白具有典型的GTP结合(GBP)结构域和2个相邻的跨膜结构,被定位在高尔基体和内质网膜上,具有运输小囊泡的功能.用人类和果蝇的Atlastin基因序列对家蚕基因组数据库进行同源搜索,鉴定得到4个同源基因,命名为BmAT1-BmATL4.生物信息学分析显示这4个基因编码的蛋白质都有GBP结构域,其中:BmATL1和BmATL2的分子质量约60kD,有2个相邻的跨膜结构;BmATL3和BmAL4的分子质量约85kD,没有跨膜结构.表达谱分析表明BmATL1和BmATL2在家蚕5龄第3天幼虫各组织都有低量表达,BmATL3和BmATL4在血细胞中特异

  7. Sequential steps of macroautophagy and chaperone-mediated autophagy are involved in the irreversible process of posterior silk gland histolysis during metamorphosis of Bombyx mori.

    Science.gov (United States)

    Shiba, Hajime; Yabu, Takeshi; Sudayama, Makoto; Mano, Nobuhiro; Arai, Naoto; Nakanishi, Teruyuki; Hosono, Kuniaki

    2016-04-15

    To elucidate the degradation process of the posterior silk gland during metamorphosis of the silkworm ITALIC! Bombyx mori, tissues collected on the 6th day after entering the 5th instar (V6), prior to spinning (PS), during spinning (SP) and after cocoon formation (CO) were used to analyze macroautophagy, chaperone-mediated autophagy (CMA) and the adenosine triphosphate (ATP)-dependent ubiquitin proteasome. Immediately after entering metamorphosis stage PS, the levels of ATP and phosphorylated p70S6 kinase protein decreased spontaneously and continued to decline at SP, followed by a notable restoration at CO. In contrast, phosphorylated AMP-activated protein kinase α (AMPKα) showed increases at SP and CO. Most of the Atg8 protein was converted to form II at all stages. The levels of ubiquitinated proteins were high at SP and CO, and low at PS. The proteasome activity was high at V6 and PS but low at SP and CO. In the isolated lysosome fractions, levels of Hsc70/Hsp70 protein began to increase at PS and continued to rise at SP and CO. The lysosomal cathepsin B/L activity showed a dramatic increase at CO. Our results clearly demonstrate that macroautophagy occurs before entering the metamorphosis stage and strongly suggest that the CMA pathway may play an important role in the histolysis of the posterior silk gland during metamorphosis. PMID:26944491

  8. A Shark Liver Gene-Derived Active Peptide Expressed in the Silkworm, Bombyx mori: Preliminary Studies for Oral Administration of the Recombinant Protein

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    Jun Li

    2013-05-01

    Full Text Available Active peptide from shark liver (APSL is a cytokine from Chiloscyllium plagiosum that can stimulate liver regeneration and protects the pancreas. To study the effect of orally administered recombinant APSL (rAPSL on an animal model of type 2 diabetes mellitus, the APSL gene was cloned, and APSL was expressed in Bombyx mori N cells (BmN cells, silkworm larvae and silkworm pupae using the silkworm baculovirus expression vector system (BEVS. It was demonstrated that rAPSL was able to significantly reduce the blood glucose level in mice with type 2 diabetes induced by streptozotocin. The analysis of paraffin sections of mouse pancreatic tissues revealed that rAPSL could effectively protect mouse islets from streptozotocin-induced lesions. Compared with the powder prepared from normal silkworm pupae, the powder prepared from pupae expressing rAPSL exhibited greater protective effects, and these results suggest that rAPSL has potential uses as an oral drug for the treatment of diabetes mellitus in the future.

  9. Molecular cloning of a cDNA for a small GTP binding protein, BRho, from the embryo of Bombyx mori and its characterization after expression and purification.

    Science.gov (United States)

    Uno, T; Nakasuji, A; Hara, W; Aizono, Y

    2000-04-01

    A cDNA clone encoding a small GTP binding protein (Brho) was isolated from an embryonic cDNA library of Bombyx mori that encoded a polypeptide with 202 amino acids sharing 60-80% similarity with the Rho1 family of GTP binding proteins. The effector site and one of the guanine nucleotide binding sites differed from other members of the Rho family. To characterize the biochemical properties of Brho, the clone was expressed in Escherichia coli as a glutathione S-transferase (GST) fusion protein. The recombinant protein was purified to homogeneity with glutathione S-Sepharose. The fusion protein bound [(35)S] GTPgammaS and [(3)H] GDP with association constants of 11x10(6) M(-1) and 6.2x10(6) M(-1), respectively. The binding of [(35)S] GTPgammaS was inhibited by GTP and GDP, but by no other nucleotides. The calculated GTP-hydrolysis activity was 89.6 m mol/min/mol of Brho. Bound [(35)S] GTPgammaS and [(3)H] GDP were exchanged with GTPgammaS most efficiently in the presence of 6 mM MgCl(2). These results suggest that Brho has a higher affinity for GTP than GDP, converts from the GTP-bound state into the GDP-bound state by intrinsic GTP hydrolytic activity, and returns to the GTP-bound state with the exchange of GDP with GTP. Arch. PMID:10737920

  10. Molecular cloning of cDNA for BRab from the brain of Bombyx mori and biochemical properties of BRab expressed in Escherichia coli.

    Science.gov (United States)

    Uno, T; Ueno, M; Nakajima, A; Shirai, Y; Aizono, Y

    1998-10-01

    From a brain cDNA library of Bombyx mori, we cloned cDNA for BRab, which encoded a 202-amino-acid polypeptide sharing 60-80% similarity with rab1 family members. To characterize its biochemical properties, cDNA for BRab was inserted into an expression vector (pGEX2T) and expressed in Escherichia coli as a glutathione S-transferase (GST) fusion protein. The recombinant protein was purified to homogeneity with glutathione S-Sepharose. The purified GST-BRab bound [35S]-GTP gamma S and [3H]-GDP with association constants of 1.5 x 10(6) M-1 and 0.58 x 10(6) M-1, respectively. The binding of [35S]-GTP gamma S was inhibited with GTP and GDP, but with no other nucleotides. The GTP-hydrolysis activity was evaluated to be 5 m mole/min/mole of BRab. In the presence of 6 mM MgCl2, bound [35S]-GTP gamma S and [3H]-GDP were exchanged with GTP gamma S most efficiently. These results suggest that BRab, having a higher affinity for GTP than GDP, converts from the GTP-bound state into the GDP-bound state by intrinsic GTP hydrolysis activity and returns to the GTP-bound state with the exchange of GDP with GTP. PMID:9836423

  11. Genome wide microarray based expression profiles associated with BmNPV resistance and susceptibility in Indian silkworm races of Bombyx mori.

    Science.gov (United States)

    Lekha, Govindaraj; Gupta, Tania; Awasthi, Arvind K; Murthy, Geetha N; Trivedy, Kanika; Ponnuvel, Kangayam M

    2015-12-01

    The molecular mechanism involved in BmNPV resistance was investigated using a genome wide microarray in midgut tissue of Indian silkworm Bombyx mori. In resistant race (Sarupat), 735 genes up-regulated and 589 genes down-regulated at 12 h post BmNPV infection. Similarly, in case of susceptible race (CSR-2), 2183 genes up-regulated and 2115 genes down-regulated. Among these, nine up-regulated and eight down-regulated genes were validated using real-time qPCR analysis. In Sarupat, vacuolar protein sorting associated, Xfin-like protein and carboxypeptidase E-like protein genes significantly up-regulated in infected midgut; prominently down-regulated genes were glutamate receptor ionotropic kainite 2-like, BTB/POZ domain and transferrin. Considerably up-regulated genes in the CSR-2 were peptidoglycan recognition protein S6 precursor and rapamycin while the conspicuous down-regulated genes were facilitated trehalose transporter and zinc transporter ZIP1-like gene. The up-regulation of genes in resistant race after BmNPV infection indicates their possible role in antiviral immune response. PMID:26376410

  12. CONSUMO E UTILIZAÇÃO DO ALIMENTO PELO BICHO-DA-SEDA (Bombyx mori L., ALIMENTADO COM DOIS CULTIVARES DE AMOREIRA EM DIFERENTES IDADES DE CORTE

    Directory of Open Access Journals (Sweden)

    Sílvia Maria Alves Gomes Dierckx

    2006-10-01

    Full Text Available O presente estudo foi desenvolvido na Unidade de Pesquisa e Desenvolvimento de Gália-SP, da APTA/SAA, no período de 31 de dezembro de 1997 a 29 de abril de 1998. Teve por objetivo avaliar, através de índices nutricionais, os efeitos da idade de corte (7, 10, 13 e 16 semanas de dois cultivares de amoreira (IZ 56/4 e Korin sobre o consumo e utilização do alimento pelo bicho-da-seda (Bombyx mori L..Com dez semanas de desenvolvimento o cultivar IZ 56/4 apresentou menor consumo e maior eficiência de utilização pelas lagartas no quinto ínstar, tendência também observada para o cultivar Korin. No geral, o cultivar Korin proporcionou as melhores características nutricionais, com menor ingestão, menor custo metabólico, maior eficiência de conversão e boa taxa de crescimento e digestibilidade. As lagartas do bicho-da-seda apresentaram boa capacidade compensatória em condições nutricionais inadequadas, de forma a manter um ótimo crescimento e desenvolvimento. PALAVRAS-CHAVE: Bicho-da-seda, idade de corte, Morus sp.

  13. New polar constituents of the pupae of the silkworm Bombyx mori L. I. Isolation and identification of methionine sulfoxide, methionine sulfone, and gamma-cyclic di-L-glutamate.

    Science.gov (United States)

    Tanaka, Ryuichiro

    2007-12-01

    In addition to serine (L:D = 68:32), methionine sulfoxide (MSO), L-methionine sulfone (L-MSO(2)), and disodium gamma-cyclic di-L-glutamate were identified in a methanol extract of Bombyx mori L. pupae. MSO was isolated in a diastereomeric mixture of L(+)- and D(+)-MSO in a ratio of 99:1. The presence of these compounds in other developmental stages, including eggs, larvae (1st, 4th, 5th, and mature 5th instar), adults, and excrement (feces and urine) was investigated. The L(+)-isomer of MSO was present in extracts of the 1st and 5th instar larvae, adults, and eggs, but was not detected in feces or urine. The D(+)-isomer was found only in pupal stage extracts, and was excreted into the meconium with L(+)-isomer. L-MSO(2) and gamma-cyclic di-L-glutamate were not detected at other insect life stages or in the insect excrement. gamma-Cyclic di-L-glutamate is thought be produced due to blockage of the glutamate synthetic pathway (glutamine synthetase) by L-MSO(2) and Mg(2+). The biochemical role of L-MSO(2) during the pupal life stage remains unknown, but importantly, the stage-specific expression suggests that it is a candidate molecule for the induction of diapause. PMID:18071251

  14. Progeny of Palmistichus elaeisis Delvare and LaSalle (Hymenoptera: Eulophidae) parasitising pupae of Bombyx mori L. (Lepidoptera: Bombycidae) of different ages; Progenie de Palmistichus elaeisis Delvare e LaSalle (Hymenoptera:Eulophidae) parasitando pupas de Bombyx mori L. (Lepidoptera:Bombycidae) de diferentes idades

    Energy Technology Data Exchange (ETDEWEB)

    Pereira, Fabricio F.; Favero, Kellen; Grance, Elizangela L.V. [Universidade Federal da Grande Dourados, MS (Brazil). Fac. de Ciencias Biologicas e Ambientais], e-mail: fabriciofagundes@ufgd.edu.br, e-mail: kellenfavero@yahoo.com.br, e-mail: eli_vargasgrance@yahoo.com.br; Zanuncio, Jose C. [Universidade Federal de Vicosa (UFV), MG (Brazil). Dept. de Biologia Animal], e-mail: zanuncio@ufv.br; Serrao, Jose E. [Universidade Federal de Vicosa (UFV), MG (Brazil), Dept. de Biologia Geral], e-mail: jeserrao@ufv.br; Oliveira, Harley N. [Embrapa Agropecuaria Oeste, Dourados, MS (Brazil)], e-mail: harley@cpao.embrapa.br

    2009-09-15

    Palmistichus elaeisis Delvare and LaSalle is a natural pupal parasitoid of eucalyptus defoliator lepidopterans and is considered a promising biocontrol agent. However, the development of efficient rearing techniques for this natural enemy are fi rst required before it can be used in biocontrol programs. Bombyx mori L. pupae are potential alternative hosts for this parasitoid mass rearing, and they are easy to rear. Therefore, we investigated the most suitable host age and the effects of parasitoid age on progeny production of P. elaeisis. B. mori pupae, 24 h-, 48 h-, 72 h- or 96 h-old were exposed to P. elaeisis females of similar age. The duration of the life cycle (egg-adult) of P. elaeisis was not affected by the age of the parasitizing female; however, the host age affected parasitoid development. The best parasitization was obtained for 72h- to 96h-old parasitoid females when offered to 48h- to 72h-old host pupae, allowing the synchronized rearing of a large number of P. elaeisis offspring. (author)

  15. The Bombyx mori Ovarian Tumor Gene Bmotu and Its Alternative Splicing%家蚕卵巢肿瘤基因Bmotu及其可变剪接

    Institute of Scientific and Technical Information of China (English)

    薛高旭; 曹广力; 张鹏杰; 张瑶瑶; 薛仁宇; 贡成良

    2011-01-01

    Ovarian tumor gene (out) plays a pivotal role in ovary development of Drosophila and mutations in out gene disrupt the normal process of oogenesis. To investigate whether silkworm (Bombyx mori) has a homologous gene similar to germ line development-related gene out in Drosophila and the isoforms produced by alternative splicing of the homologous gene, four Bmotu cDNA segments with varied lengths from silkworm testis were obtained using RT-PCR based on in silico cloning (GenBank accession No. HQ831341, HQ831342, HQ999998 and HQ831343). Three of them bear stop codons which would terminate translation prematurely due to nonsense mutation. Nevertheless, only one Bmotu cDNA fragment was obtained from ovary. Sequence of this fragment is identical to the 5' end of fragment HQ831343 which was amplified from testis. These results indicated that Bmotu gene has alternative splicing and that diverse alternatively spliced isoforms exist in male and female silkworms. Bioinformatics analysis discovered that proteins encoded by Bmotu are similar to Drosophila Out in structure which contains a cysteine proteinase domain (Out domain), a Tudor domainand a proline-rich motif. The obtained data are conducive to further investigation on mechanism of silkworm germ line development.%已知卵巢肿瘤基因(ovarian tumor gene,otu)在果蝇(Drosophila melanogaster)卵巢发育过程中发挥极其重要的作用,该基因突变会扰乱卵子形成的正常过程.为探究家蚕(Bombyx mori)是否具有类似果蝇生殖发育相关基因otu的同源基因,以及基因存在的可变剪接形式,在电子克隆的基础上,应用RT-PCR从家蚕精巢组织中获得了4条不同长度的Bmotu cDNA片段(GenBank登录号:HQ831341,HQ831342,HQ999998,HQ831343),其中3条由于无义突变导致翻译提前终止;而从卵巢组织中仅获得了1条Bmotu cDNA片段,该片段序列与精巢中扩增的登录号为HQ831343片段序列的5’端相同.结果表明Bmotu基因存在可变剪接,

  16. Studies on resistance mechanism of Bombyx mori against Bacillus thuringiensis insecticidal Cry1Ac toxin-Role of Cry1Ac binding proteins localizing on peritrophic and epithelial cell membranes of midgut-

    OpenAIRE

    Shitomi, Yasuyuki; 蔀, 泰幸

    2006-01-01

    Bombyx mori, hybrid Shunrei x Shogetsu, is susceptible to Cry1Aa and insensitive to Cry1Ac. Toxicity of Cry toxin can be correlated with the presence of a specific receptor in midgut epithelial cell membrane in insect. In surface plasmon resonance (SPR) and ligand blot analysis, however, many kinds of brush border membrane vesicle (BBMV) proteins were shown to bind to Cry1Ac with almost equal intensity in both resistance and susceptible insects. This suggests that majority of the bindings bet...

  17. EFFECT OF AQUEOUS MACERATIVES OF SEED POWDER OF SYZIGIUM CUMINI (L) ON THE PROTEIN CONTENTS AND MID GUT ENZYME ACTIVITIES IN THE FIFTH INSTAR LARVAE OF SILK WORM, BOMBYX MORI (L) (RACE: PM X CSR2)

    OpenAIRE

    Tejas C Korade, Manali T Patil, Kavita H Nimbalkar

    2013-01-01

    Different concentrations (5.0 ppm; 10.0 ppm; 20.0 ppm & 50.0 ppm) of the aqueous solution of seed powder of Syzigium cumini (L) concentrations was used to treat the leaves of mulberry & fed to the fifth instar larvae of polyvoltine, crossbreed silkworm, Bombyx mori (L) (Race : PM x CSR2 ) for first three days; second day & third day; third day (only). The larvae fed with untreated & water treated leaves were also maintained. Bioassays of proteins (S.P. & T.P.) & enzyme...

  18. LIM-homeodomain transcription factor Awh is a key component activating all three fibroin genes, fibH, fibL and fhx, in the silk gland of the silkworm, Bombyx mori

    OpenAIRE

    Kimoto, Mai; Tsubota, Takuya; Uchino, Keiro; Sezutsu, Hideki; Takiya, Shigeharu

    2015-01-01

    In the silkworm Bombyx mori, three fibroin genes, fibroin-heavy-chain (fibH), fibroin-light-chain (fibL) and fibrohexamerin (fhx), are coexpressed only in the posterior silk gland (PSG) cells, while the sericin genes encoding silk glue proteins are expressed in the middle silk gland (MSG) cells. Silk gland factor-2 (SGF-2) is a PSG-specific activator complex of fibH, composed of a LIM-homeodomain protein, Awh, and its cofactors, Ldb and Lcaf. We investigated whether SGF-2 can activate other f...

  19. The lipopolysaccharide-binding protein participating in hemocyte nodule formation in the silkworm Bombyx mori is a novel member of the C-type lectin superfamily with two different tandem carbohydrate-recognition domains.

    Science.gov (United States)

    Koizumi, N; Imamura, M; Kadotani, T; Yaoi, K; Iwahana, H; Sato, R

    1999-01-25

    We recently isolated and characterized the lipopolysaccharide (LPS)-binding protein, BmLBP, from the larval hemolymph of the silkworm Bombyx mori. BmLBP is a pattern recognition molecule that recognizes the lipid A portion of LPS and participates in a cellular defense reaction. This paper describes the cDNA cloning of BmLBP. The deduced amino acid sequence of BmLBP revealed that BmLBP is a novel member of the C-type lectin superfamily with a unique structural feature that consists of two different carbohydrate-recognition domains in tandem, a short and a long form. PMID:9989592

  20. Hierarchical clustering of 54 races and strains of the mulberry silkworm, Bombyx mori L: Significance of biochemical parameters.

    Science.gov (United States)

    Chatterjee, S N; Datta, R K

    1992-12-01

    A detailed analysis was undertaken to test the efficacy of hierarchical agglomerative clustering (UPGMA method) in grouping the races and strains of the mulberry silkworm, Bombyx moti L., and to ascertain the importance of biochemical parameters in the clustering process. The analysis was based on data from two rearing seasons with 54 selected races/strains of different geographic origin and varying yield potentials. The results indicate that seven clusters can be realised with yield parameters alone, whereas the inclusion of biochemical parameters in clustering resulted into two broad groups: one having all the breeds with high cocoon weight and shell weight, the other having all the low-yielding silkworm strains both from India and from other countries. Further sub-grouping under these two groups highlights genetical differences associated with the differentiation of various groups of races in temperate and tropical areas as well as their significance for silkworm breeding. Estimates of all ten variables were further subjected to 'quick clustering' and the results showed that cluster 5, constituted by 38 lowyielding strains of India, China and Europe, had the highest values of the final cluster centre for amylase and the effective rate of rearing (ERR), while clusters 1 and 4 had the highest values for invertase and alkaline phosphatase. The evolutionary aspect of the genetic channelisation of silkworm races from various countries is discussed against the background of differences in the biochemical parameters and yield variables. PMID:24197452

  1. DNA Synthesis in the Giant Nuclei of Insects - Control Machinery and Structures Observed in the Silk-Producing Gland of Bombyx Mori

    International Nuclear Information System (INIS)

    The existence in many insect organs of giant nuclei without visible chromosomes raises the question of possible homologies between the chromatin structures of these nuclei and those of polytene nuclei or common euploid cells. Studies have been made of the nuclei in the silk-producing gland of Bombyx mori. The DNA synthesis is cyclic. During the third stage there are three successive synthesis cycles, which appear to be relatively autonomous in the individual nuclei. For more than 24 hours after moulting, however, synthesis is greatly reduced; moulting factors thus cause synchronization of all the nuclei. This leads to the conclusion that the triggering of a synthesis cycle is controlled by general factors external to the cell. At the end of larval development, DNA synthesis is suspended at the moment when large-scale secretion of silk begins. Evaluation of the pool of endogenic precursors of DNA shows that it is considerably reduced at the end of the DNA synthesis period. The hypothesis proposed is that large-scale synthesis of fibroin requires polarization of the metabolism, hence the depletion of the nucleotide pool and the end of DNA synthesis. DNA synthesis within a single nucleus is to some extent asynchronic. In particular, a well-defined, delayed-synthesis structure visible only in the female seems to be a possible homologue of a sex chromosome. Other asynchronisms are also apparent, though less clearly. Functional studies thus allow the supposition that in the giant nucleus replication units retain an individuality comparable to that of a polytene chromosome. These observations together lead to the conclusion that a nucleus in the silk-producing gland has physiological and structural characteristics similar to those of a polytene nucleus, differing from it essentially in the lesser degree of condensation of its structures. (author)

  2. The arginine residue within the C-terminal active core of Bombyx mori pheromone biosynthesis-activating neuropeptide (PBAN is essential for receptor binding and activation

    Directory of Open Access Journals (Sweden)

    Takeshi eKawai

    2012-03-01

    Full Text Available In most lepidopteran insects, the biosynthesis of sex pheromones is regulated by pheromone biosynthesis activating neuropeptide (PBAN. Bombyx mori PBAN (BomPBAN consists of 33 amino acid residues and contains a C-terminus FSPRLamide motif as the active core. Among neuropeptides containing the FXPRLamide motif, the arginine (Arg, R residue two positions from the C-terminus is highly conserved across several neuropeptides, which can be designated as RXamide peptides. The purpose of this study was to reveal the role of the Arg residue in the BomPBAN active core. We synthesized a ten-residue peptide corresponding to the C-terminal part of BomPBAN with a series of point mutants at the 2nd position (ie, Arg from the C-terminus, termed the C2 position, and measured their efficacy in stimulating Ca2+ influx in insect cells concomitantly expressing a fluorescent PBAN receptor chimera (PBANR-EGFP and loaded with the fluorescent Ca2+ indicator, Fura Red-AM. PBAN analogs with the C2 position replaced with alanine (Ala, A, aspartic acid (Asp, D, serine (Ser, S or L-2-aminooctanoic acid (Aoc decreased PBAN-like activity. RC2A (SKTRYFSPALamide and RC2D (SKTRYFSPDLamide had the lowest activity and could not inhibit the activity of PBAN C10 (SKTRYFSPRLamide. We also prepared Rhodamine Red-labeled PBAN analogs of the mutants and examined their ability to bind PBANR. In contrast to 100 nM Rhodamine Red-PBAN C10, none of the mutants at the same concentration exhibited PBANR binding. Taken together, our results demonstrate that the C2 Arg residue in BomPBAN is essential for PBANR binding and activation.

  3. The p10 gene of Bombyx mori nucleopolyhedrosis virus encodes a 7.5-kDa protein and is hypertranscribed from a TAAG motif

    Indian Academy of Sciences (India)

    Vikas B. Palhan; Karumathil P. Gopinathan

    2000-08-01

    In baculovirus-based high-level expression of cloned foreign genes, the viral very late gene promoters of polyhedrin (polh) and p10 are extensively exploited. Here we report the cloning and characterization of the p10 gene from a local isolate of Bombyx mori nucleopolyhedrosis virus (BmNPV). The gene harbours a 213-bp open reading frame encoding a protein of 70 amino acids with a predicted molecular mass of 7.5 kDa. The BmNPV p10 showed deletion of a single A at +210 nucleotide compared to the prototype baculovirus, Autographa californica multinucleocapsid nucleopolyhedrosis virus (AcMNPV), p10 gene, resulting in a translational frameshift to generate a termination codon and consequently a truncated polypeptide instead of the 10-kDa protein. This protein P7.5 from BmNPV has a putative leucine zipper dimerization motif towards the N-terminal end and the central nuclear disintegration domain but the carboxy-terminal domain implicated in protein association for fibrillar structure formation is absent. Phylogenetic analysis revealed that p10 is highly conserved among baculoviruses and the BmNPV strains are more closely related to AcMNPV than other baculoviruses. The transcription of p10 is regulated in a temporal manner, reaching maximal levels by 72 h post-infection. RNAase protection and primer extension analysis mapped the transcription start sites at $-70$ and $-71$ nt with respect to the ATG, within the conserved baculovirus late gene motif T\\underline{AA}G. The upstream region showed complete homology to the strong promoter of the AcMNPV p10, suggesting that this promoter from BmNPV could also be exploited for high-level expression of cloned foreign genes in silkworm cells or larvae.

  4. Translational enhancement of recombinant protein synthesis in transgenic silkworms by a 5'-untranslated region of polyhedrin gene of Bombyx mori Nucleopolyhedrovirus.

    Science.gov (United States)

    Iizuka, Masashi; Tomita, Masahiro; Shimizu, Katsuhiko; Kikuchi, Yutaka; Yoshizato, Katsutoshi

    2008-06-01

    Previously, we established a method to produce recombinant proteins (r-proteins) in cocoons of germline transgenic silkworms, and showed that a step(s) in post-transcription processes was rate-limiting in obtaining a high yield of r-proteins. In this study, we examined whether the 5'-untranslated region (5'-UTR) of the polyhedrin gene (pol) of nucleopolyhedrovirus (NPV) has a translational enhancer activity in the r-protein expression by middle silk gland (MSG) cells of silkworm Bombyx mori (Bm). Sericin 1 gene (ser1) promoter-driven transformation vectors were constructed in which pol5'-UTRs of NPVs isolated from four different species, Bm, Spodoptera frugiperda, Ectropis oblique, and Malacosoma neustria, were each placed upstream of a reporter gene. Transient expression assays in MSGs showed that these pol5'-UTRs all enhanced the protein expression of reporter genes, and the pol5'-UTR of Bm NPV (pol5'-UTR/Bm) was the most effective among them. Thus, transgenic silkworms were generated, which bore the ser1 promoter-driven His-tagged secretory EGFP (sEGFP-His) gene under the control of pol5'-UTR/Bm. The synthesis of sEGFP-His proteins in MSGs of the transgenic worms was approximately 1.5-fold higher than that in those bearing null vectors. However, its mRNA expression levels were 67% of the control worms, indicating that the pol5'-UTR/Bm specifically enhanced the translational level. In conclusion, pol5'-UTR/Bm increased the yield of r-protein production in transgenic silkworms by enhancing the translational activity and this 5'-UTR could be useful for the mass production of r-proteins in germline transgenic silkworms. PMID:18640598

  5. Intermittent expression of BmFTZ-F1, a member of the nuclear hormone receptor superfamily during development of the silkworm Bombyx mori.

    Science.gov (United States)

    Sun, G C; Hirose, S; Ueda, H

    1994-04-01

    BmFTZ-F1 is a sequence-specific DNA-binding factor in the silkworm Bombyx mori sharing similar biochemical characteristics with Drosophila FTZ-F1, a member of the nuclear hormone receptor superfamily. Using DNA sequence homology with FTZ-F1 and information on tryptic peptide sequences of BmFTZ-F1, we isolated a cDNA encoding for BmFTZ-F1. Amino acid sequences in the zinc finger DNA-binding region and the putative ligand-binding domain of BmFTZ-F1 showed strong similarity to not only FTZ-F1 but also its mammalian homologues, LRH-1, ELP, and Ad4BP, suggesting the importance of each region for the function of these proteins. Northern blot analyses of RNA isolated from the middle and posterior silk glands and fat bodies showed that a 6.1-kb BmFTZ-F1 mRNA is present in all tissues so far examined. Expression of BmFTZ-F1 mRNA is intermittent, being high during larval molting and both the larval-pupal and the pupal-adult transformations. Injection of 20-hydroxyecdysone at the third day of the 5th instar larvae induced BmFTZ-F1 mRNA in the posterior silk gland after 24 hr. When 5th instar silk glands were cultured in vitro, BmFTZ-F1 mRNA was induced by a 6-hr exposure to 20-hydroxyecdysone followed by 6 hr in hormone-free medium. These results suggest that BmFTZ-F1 is inducible by decline in the ecdysteroid titer and may play an important role in the development of the silkworm as a transcription factor. PMID:8150206

  6. Functional characterization of Bombyx mori nucleopolyhedrovirus late gene transcription and genome replication factors in the non-permissive insect cell line SF-21

    International Nuclear Information System (INIS)

    We compared the abilities of late gene transcription and DNA replication machineries of the baculoviruses Autographa californica nucleopolyhedrovirus (AcMNPV) and Bombyx mori NPV (BmNPV) in SF-21 cells, an insect-derived cell line permissive for AcMNPV infection. It has been well established that 19 AcMNPV late expression factors (lefs) stimulate substantial levels of late gene promoter activity in SF-21 cells. Thus, we constructed a set of clones containing the BmNPV homologs of the AcMNPV lefs under control of the constitutive Drosophila heat shock 70 protein promoter and tested their ability to activate an AcMNPV late promoter-reporter gene cassette in SF-21 cells. We tested the potential of individual or predicted functional groups of BmNPV lefs to successfully replace the corresponding AcMNPV gene(s) in transient late gene expression assays. We found that most, but not all, BmNPV lefs were able to either fully or partially substitute for the corresponding AcMNPV homolog in the context of the remaining AcMNPV lefs with the exception of BmNPV p143, ie-2, and p35. BmNPV p143 was unable to support late gene expression or be imported into the nucleus of cells in the presence of the AcMNPV or the BmNPV LEF-3, a P143 nuclear shuttling factor. Our results suggest that host-specific factors may affect the function of homologous proteins

  7. Molecular tracing of white muscardine in the silkworm, Bombyx mori (Linn.) II. Silkworm white muscardine is not caused by artificial release or natural epizootic of Beauveria bassiana in China.

    Science.gov (United States)

    Chen, Xue; Huang, Cui; He, Lingmin; Zhang, Shengli; Li, Zengzhi

    2015-02-01

    The fungal pathogen Beauveria bassiana causes serious economic losses in sericulture. Its origin is usually attributed to the release of B. bassiana insecticides against pine caterpillars (Dendrolimus punctuatus). In the present study, 488 B. bassiana isolates obtained from silkworm (Bombyx mori) collected from 13 Chinese provinces, and 327 B. bassiana isolates obtained from D. punctatus collected from 9 provinces, were analyzed for population genetic structure using the ISSR technique based on genetic distance. A UPGMA dendrogram clustered them into three independent clades: two B. mori clades and one D. punctatus clade. A 3-D principal component analysis further divided them into two completely independent host groups, revealing high host-specificity. This suggested that white muscardine occurring in B. mori populations throughout southern China was not caused by any B. bassiana strain either naturally prevailing in D. punctatus populations or by any strain artificially released as a fungal insecticide against D. punctatus. We further investigated the genetic differentiation coefficient Gst and gene flow between B. mori-pathogenic and D. punctatus-pathogenic B. bassiana isolates from across China and from five provinces inhabited by both B. mori and D. punctatus. The Gst value across China was computed as 0.410, while the values of the five provinces ranged from 0.508 to 0.689; all above 0.25, which is the threshold for significant genetic differentiation. This suggests that B. bassiana strains isolated from the two different hosts maintained their respective heredity without a convergent homogenization trend, and reduces the possibility that the host range of the caterpillar isolates could expand and enhance their virulence in B. mori. These findings indicate that the use of B. bassiana does not threaten the safety of sericulture. PMID:25541121

  8. Lipopolysaccharide-binding protein of Bombyx mori participates in a hemocyte-mediated defense reaction against gram-negative bacteria.

    Science.gov (United States)

    Koizumi, N; Imai, Y; Morozumi, A; Imamura, M; Kadotani, T; Yaoi, K; Iwahana, H; Sato, R

    1999-09-01

    BmLBP is a lipopolysaccharide-binding protein in B. mori and participates in bacterial clearance in vivo. Here, we investigated the function of BmLBP more specifically. More than 90% of injected gram-negative rough strains to which BmLBP binds were removed from the plasma within 30 min post-injection, whereas it required 8h for the clearance of smooth strains to which BmLBP does not bind. Observation of the hemocoel after the injection of Escherichia coli rough strain showed that melanized nodules were formed at 30 min post-injection when the clearance of injected E. coli cells had occurred. Fluorescence microscope observation revealed that E. coli cells were actually trapped in the nodules formed in vivo. Furthermore, plasma pre-treated E. coli rough cells (BmLBP bound) added to hemocytes isolated in vitro caused vigorous hemocyte aggregations with the bacteria, while plasma pre-treated smooth cells did not. The formation of aggregates was inhibited by anti-BmLBP serum pre-treatment, suggesting that BmLBP causes the clearance of bacteria by promoting hemocyte nodule formation. PMID:12770298

  9. Single amino acid insertions in extracellular loop 2 of Bombyx mori ABCC2 disrupt its receptor function for Bacillus thuringiensis Cry1Ab and Cry1Ac but not Cry1Aa toxins.

    Science.gov (United States)

    Tanaka, Shiho; Miyamoto, Kazuhisa; Noda, Hiroaki; Endo, Haruka; Kikuta, Shingo; Sato, Ryoichi

    2016-04-01

    In a previous report, seven Cry1Ab-resistant strains were identified in the silkworm, Bombyx mori; these strains were shown to have a tyrosine insertion at position 234 in extracellular loop 2 of the ABC transporter C2 (BmABCC2). This insertion was confirmed to destroy the receptor function of BmABCC2 and confer the strains resistance against Cry1Ab and Cry1Ac. However, these strains were susceptible to Cry1Aa. In this report, we examined the mechanisms of the loss of receptor function of the transporter by expressing mutations in Sf9 cells. After replacement of one or two of the five amino acid residues in loop 2 of the susceptible BmABCC2 gene [BmABCC2_S] with alanine, cells still showed susceptibility, retaining the receptor function. Five mutants with single amino acid insertions at position 234 in BmABCC2 were also generated, resulting in loop 2 having six amino acids, which corresponds to replacing the tyrosine insertion in the resistant BmABCC2 gene [BmABCC2_R(+(234)Y)] with another amino acid. All five mutants exhibited loss of function against Cry1Ab and Cry1Ac. These results suggest that the amino acid sequence in loop 2 is less important than the loop size (five vs. six amino acids) or loop structure for Cry1Ab and Cry1Ac activity. Several domain-swapped mutant toxins were then generated among Cry1Aa, Cry1Ab, and Cry1Ac, which are composed of three domains. Swapped mutants containing domain II of Cry1Ab or Cry1Ac did not kill Sf9 cells expressing BmABCC2_R(+(234)Y), suggesting that domain II of the Cry toxin is related to the interaction with the receptor function of BmABCC2. This also suggests that different reactions against Bt-toxins in some B. mori strains, that is, Cry1Ab resistance or Cry1Aa susceptibility, are attributable to structural differences in domain II of Cry1A toxins. PMID:26928903

  10. Crystal structure of silkworm Bombyx mori JHBP in complex with 2-methyl-2,4-pentanediol: plasticity of JH-binding pocket and ligand-induced conformational change of the second cavity in JHBP.

    Directory of Open Access Journals (Sweden)

    Zui Fujimoto

    Full Text Available Juvenile hormones (JHs control a diversity of crucial life events in insects. In Lepidoptera which major agricultural pests belong to, JH signaling is critically controlled by a species-specific high-affinity, low molecular weight JH-binding protein (JHBP in hemolymph, which transports JH from the site of its synthesis to target tissues. Hence, JHBP is expected to be an excellent target for the development of novel specific insect growth regulators (IGRs and insecticides. A better understanding of the structural biology of JHBP should pave the way for the structure-based drug design of such compounds. Here, we report the crystal structure of the silkworm Bombyx mori JHBP in complex with two molecules of 2-methyl-2,4-pentanediol (MPD, one molecule (MPD1 bound in the JH-binding pocket while the other (MPD2 in a second cavity. Detailed comparison with the apo-JHBP and JHBP-JH II complex structures previously reported by us led to a number of intriguing findings. First, the JH-binding pocket changes its size in a ligand-dependent manner due to flexibility of the gate α1 helix. Second, MPD1 mimics interactions of the epoxide moiety of JH previously observed in the JHBP-JH complex, and MPD can compete with JH in binding to the JH-binding pocket. We also confirmed that methoprene, which has an MPD-like structure, inhibits the complex formation between JHBP and JH while the unepoxydated JH III (methyl farnesoate does not. These findings may open the door to the development of novel IGRs targeted against JHBP. Third, binding of MPD to the second cavity of JHBP induces significant conformational changes accompanied with a cavity expansion. This finding, together with MPD2-JHBP interaction mechanism identified in the JHBP-MPD complex, should provide important guidance in the search for the natural ligand of the second cavity.

  11. High-titer preparation of Bombyx mori nucleopolyhedrovirus (BmNPV displaying recombinant protein in silkworm larvae by size exclusion chromatography and its characterization

    Directory of Open Access Journals (Sweden)

    Tanaka Shigeyasu

    2009-06-01

    Full Text Available Abstract Background Budded baculoviruses are utilized for vaccine, the production of antibody and functional analysis of transmembrane proteins. In this study, we tried to produce and purify the recombinant Bombyx mori nucleopolyhedrovirus (rBmNPV-hPRR that displayed human (prorenin receptor (hPRR connected with FLAG peptide sequence on its own surface. These particles were used for further binding analysis of hPRR to human prorenin. The rBmNPV-hPRR was produced in silkworm larvae and purified from its hemolymph using size exclusion chromatography (SEC. Results A rapid method of BmNPV titer determination in hemolymph was performed using quantitative real-time PCR (Q-PCR. A correlation coefficient of BmNPV determination between end-point dilution and Q-PCR methods was found to be 0.99. rBmNPV-hPRR bacmid-injected silkworm larvae produced recombinant baculovirus of 1.31 × 108 plaque forming unit (pfu in hemolymph, which was 2.8 × 104 times higher than transfection solution in Bm5 cells. Its purification yield by Sephacryl S-1000 SF column chromatography was 264 fold from larval hemolymph at 4 days post-injection (p.i., but 35 or 39 fold at 4.5 or 5 days p.i., respectively. Protein patterns of rBmNPV-hPRR purified at 4 and 5 days were the same and ratio of envelope proteins (76, 45 and 35 kDa to VP39, one of nucleocapsid proteins, increased at 5 days p.i. hPRR was detected in only purified rBmNPV-hPRR at 5 days p.i.. Conclusion The successful purification of rBmNPV-hPRR indicates that baculovirus production using silkworm larvae and its purification from hemolymph by Sephacryl S-1000 SF column chromatography can provide an economical approach in obtaining the purified BmNPV stocks with high titer for large-scale production of hPRR. Also, it can be utilized for further binding analysis and screening of inhibitors of hPRR.

  12. 29种农药对家蚕的急性毒性评价%An Evaluation on Acute Toxicity of 29 Pesticides to Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    张骞; 姜辉; 肖斌; 崔新倩; 王开运

    2011-01-01

    采用食下毒叶法,测定29种农药对家蚕的急性毒性,结果表明:阿维菌素、多杀菌素、氯氰菊酯·毒死蜱和吡虫啉对家蚕96h的LC50分别为0.0020、0.0891、0.2145和0.2729 mg/L,属剧毒级农药;阿维菌素·苏云金杆菌、丁硫克百威、敌百虫、除虫菊素·苦皮藤素、丙溴磷、苏云金杆茵·毒死蜱和烟碱对家蚕96h的LC50分别为0.6921、0.7696、0.8502、0.7449、2.5134、1.7327和7.2604 m/L,属高毒级农药;其余药剂均属中、低毒级农药.以上剧毒级或高毒级的农药应当禁止在蚕区使用.吡蚜酮对家蚕的毒性低,但在蚕区是否能使用低毒级农药,还须考虑药剂的残毒期、慢性毒性和传染性等因素.%The acute toxicity of 29 pesticides to silkworm, Bombyx mori, was evaluated in laboratory by using the leaf dipping method. The results indicated that LC50 of abamectin, spinosad, cypermethrin· chlorpyrifos, and imidacloprid was 0.002 0, 0.089 1,0. 214 5, and 0. 272 9 mg/L respectively, being pesticides of virulent toxicity grade. The LC50 of abamectin· Bacillus thuringiensis, carbosulfan, trichlorfon, pyrethrins ·celastrusangulatus, profenofos, Bacillus thuringiensis·chlorpyrifos, and nicotine was 0. 692 1,0. 769 6, 0. 850 2, 0. 744 9, 2. 513 4, 1. 732 7, 7. 260 4 mg/L respectively, being pesticides of high toxicity grade. All the other pesticides were of moderate or Iow toxicity grade. Pesticides of virulent and high toxicity grade should be forbidden to use in silkworm rearing areas. Pymetrozine is of Iow toxicity grade. Yet the residual toxicity period, chronic toxicity and infectivity should be considered if pesticides of Iow toxicity grade are to be used in silkworm rearing areas.

  13. Induction of male sterility through gamma radiation in the uzifly, Exorista sorbillans (Diptera: Tachinidae), a serious parasitoid of silkworm, Bombyx mori (Lepidoptera: Bombycidae)

    International Nuclear Information System (INIS)

    Full text: The paper deals with the study on the induction of male sterility in Exorista sorbillans (Diptera: Tachinidae), which is a serious pest of the popular mulberry silkworm, Bombyx mori (Lepidoptera: Bombycidae). The endoparasitoid inflicts considerable damage to the silk industry by large-scale parasitisation of the silkworm, leading to heavy losses ranging from 30-70%. The pest, which was unknown till a few decades ago in peninsular India, has firmly established itself due to its reproduction and locomotor activities. Induction of male sterility through chemosterilants has been attempted to limit the reproductive porential of the fly population. The present study deals with induction of male sterility through gamma radiation. The uzi maggots were treated with different doses of gamma radiation and their effects on maggot, pupal and adult mortality, longevity, fecundity and cytology of uzifly were investigated in detail. The results show that the dosage of 11 Gy is relatively more effective in the induction of male sterility on large scale. When the untreated female was crossed with the irradiated male, the number of eggs laid was normal indicating no reduction of fecundity. However, none of the eggs hatched, probably due to the fact that the exposed maggots contain germ cells that were irradiated before gametogenesis. This resulted in a complete nuclear change in a germ cell, which prevents it from maturing or participating in zygote formation. In addition, it was found that the irradiation had an effect on the size and shape of the testis. By releasing these irradiated sterile flies, which have a high survival and vigour, we can ensure the reduction of the natural population. The cytological basis of induced male sterility has been studied through chromosome preparation of testis and ovaries of irradiated pupae. Chromosomal aberrations like deletions and fragmentations were observed. The ultrastructural variation in irradiated spermatids and abnormalities

  14. The expression analysis of cysteine proteinase-like protein in wild-type and nm2 mutant silkworm (Lepidoptera: Bombyx mori).

    Science.gov (United States)

    Wu, Fan; Kang, Lequn; Wang, Pingyang; Zhao, Qiaoling

    2016-07-15

    The mutant of non-molting in the 2nd instar (nm2) is a recently discovered mutant of Bombyx mori. The mutant cannot molt and exuviate and died successively in premolting of 2nd instar. In this study, two dimensional gel electrophoresis (2-DE) was performed to screen the differential expression of epidermis proteins in pre-molting larvae of 2nd instar between the wild-type and nm2 mutant. Interestingly, a cysteine proteinase-like (BmCP-like) protein in nm2 was significantly higher than that of the wild-type. The transcription profiles of BmCP-like gene were investigated by quantitative real-time PCR (qRT-PCR), and the result revealed that BmCP-like mRNA was remarkably higher in nm2 than that of the wild-type. The transcription level of BmCP-like was high in the epidermis while low in the midgut and hemocytes, and fluctuate with development, while the highest in the newly molted larvae of 3rd and lowest in the pre-molting of the 1st and 2nd instar. The body of injected BmCP-like RNAi of 2nd larvae formed a dark spots around the injection place. These results suggested the BmCP-like gene play a key role in the degradation of the cuticle and epidermis layer during molting of 1st and 2nd instar silkworm. Furthermore, the ORF of BmCP-like gene in nm2 was the same to the wild-type. These studies give us a hint that BmCP-like gene maybe not the major gene responsible for nm2, but BmCP-like gene might participate in the immune systems of silkworm, and the upregulation of BmCP-like transcription in the nm2 mutant might be induced by the disadvantages that limit the growth and development of silkworm in order to survive. PMID:27080953

  15. Molecular diagnosis for the silk worm Bombyx Mori L. Viral and bacterial diseases in the irradiated and non-irradiated individuals

    International Nuclear Information System (INIS)

    Genetic maps for the Bombyx Mori infectious flacherrie virus (BmlFV) causing flacherrie (Fl) disease and nuclear polyhedrosis virus (BmNPV) causing grasserie (Gr) disease were built up in an attempt to diagnose diseases early in young larval stages. For the non-irradiated and irradiated viral RNA of IFV, no amplification was obtained by using RT-PCR and RAPD-PCR techniques. HcoRI, EcoRV, BamHI, Hind III and BamHI restriction enzymes were used to digest the non-irradiated and irradiated viral DNA of BmNPV. It was found that, the two viral DNA samples were genetically different; the similarity indexes were 0.14, 0, 0, 0.18 and 0.15, respectively. At the biochemical level, native protein electrophoresis showed 4 and 3 new proteins in non-irradiated and irradiated Fl diseased larvae, respectively, while Gr diseased larvae showed 1 and 3 new protein types. The similarity index (S.I) between all the tested samples was not exceeded 44%. For lipoprotein pattern, 2 and 3 new lipoprotein types were appeared due to Fl disease in the non-irradiated and irradiated haemolymph samples, respectively, while Gr disease showed 3 new lipoproteins in the non-irradiated samples only. The highest S.I recorded was 56%. Glycoprotein pattern revealed 3 and 5 new glycoprotein types appeared due to Fl disease while Gr disease showed 4 and 6 new types in the non-irradiated and irradiated samples, respectively. The highest S.I was 77%. Fractionated protein with SDS revealed 2 common bands shared between the tested samples with Rf values 0.28 and 0.71. Fl disease increased the number of protein bands with the appearance of 5 and 4 new proteins types. Gr disease reduced the total number of proteins with the appearance of 2 and 3 new types. The highest S.I was 59%. Both diseases and irradiation may be mutagenic through the epigenetic level in silkworm larvae leading to death. Thus, the results of the biochemical and genetic characterization of IFV and BmNPV enable us to conclude that the

  16. Construction and application of an electronic spatiotemporal expression profile and gene ontology analysis platform based on the EST database of the silkworm, Bombyx mori.

    Science.gov (United States)

    Gan, Li-Ping; Zhang, Wen-Yu; Niu, Yan-Shan; Xu, Li; Xi, Jian; Ji, Ming-Ming; Xu, Shi-Qing

    2010-01-01

    An Expressed Sequence Tag (EST) is a short sub-sequence of a transcribed cDNA sequence. ESTs represent gene expression and give good clues for gene expression analysis. Based on EST data obtained from NCBI, an EST analysis package was developed (apEST). This tool was programmed for electronic expression, protein annotation and Gene Ontology (GO) category analysis in Bombyx mori (L.) (Lepidoptera: Bombycidae). A total of 245,761 ESTs (as of 01 July 2009) were searched and downloaded in FASTA format, from which information for tissue type, development stage, sex and strain were extracted, classified and summed by running apEST. Then, corresponding distribution profiles were formed after redundant parts had been removed. Gene expression profiles for one tissue of different developmental stages and from one development stage of the different tissues were attained. A housekeeping gene and tissue-and-stage-specific genes were selected by running apEST, contrasting with two other online analysis approaches, microarray-based gene expression profile on SilkDB (BmMDB) and EST profile on NCBI. A spatio-temporal expression profile of catalase run by apEST was then presented as a three-dimensional graph for the intuitive visualization of patterns. A total of 37 query genes confirmed from microarray data and RT-PCR experiments were selected as queries to test apEST. The results had great conformity among three approaches. Nevertheless, there were minor differences between apEST and BmMDB because of the unique items investigated. Therefore, complementary analysis was proposed. Application of apEST also led to the acquisition of corresponding protein annotations for EST datasets and eventually for their functions. The results were presented according to statistical information on protein annotation and Gene Ontology (GO) category. These all verified the reliability of apEST and the operability of this platform. The apEST can also be applied in other species by modifying some

  17. Transcriptome Sequencing and Positive Selected Genes Analysis of Bombyx mandarina

    OpenAIRE

    Tingcai Cheng; Bohua Fu; Yuqian Wu; Renwen Long; Chun Liu; Qingyou Xia

    2015-01-01

    The wild silkworm Bombyx mandarina is widely believed to be an ancestor of the domesticated silkworm, Bombyx mori. Silkworms are often used as a model for studying the mechanism of species domestication. Here, we performed transcriptome sequencing of the wild silkworm using an Illumina HiSeq2000 platform. We produced 100,004,078 high-quality reads and assembled them into 50,773 contigs with an N50 length of 1764 bp and a mean length of 941.62 bp. A total of 33,759 unigenes were identified, wi...

  18. Storage protein-2 as a dependable biochemical index for screening germplasm stocks of the silkworm Bombyx mori (L.

    Directory of Open Access Journals (Sweden)

    Jingade H. Anuradha

    2012-09-01

    Full Text Available Storage protein (SP-2 variation was investigated among selected silkworm germplasm stocks representing two major potential sericulture areas of India. The expression levels of storage protein varied among them, as seen in Sodium Dodecylsulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE, which correlated with their geographical origin. The storage protein variation is an inter origin variability and this differential expression of the protein is helpful to tag the robustness of the breed/race associated with parentage and their origin. Present study revealed that silkworm races/breeds viz., LMO, Kolar Gold and A4e possess higher protein content among the races studied. This may be correlated with their robustness reflecting higher survival rate in the varied environments prevailing in the tropical zone. Such identified races can be conserved as storage protein rich genetic stocks for their maximal genetic potentials and high-grade silk productivity.

  19. Identification and Expression Profiling of the BTB Domain-Containing Protein Gene Family in the Silkworm, Bombyx mori

    Directory of Open Access Journals (Sweden)

    Daojun Cheng

    2014-01-01

    Full Text Available The BTB domain is a conserved protein-protein interaction motif. In this study, we identified 56 BTB domain-containing protein genes in the silkworm, in addition to 46 in the honey bee, 55 in the red flour beetle, and 53 in the monarch butterfly. Silkworm BTB protein genes were classified into nine subfamilies according to their domain architecture, and most of them could be mapped on the different chromosomes. Phylogenetic analysis suggests that silkworm BTB protein genes may have undergone a duplication event in three subfamilies: BTB-BACK-Kelch, BTB-BACK-PHR, and BTB-FLYWCH. Comparative analysis demonstrated that the orthologs of each of 13 BTB protein genes present a rigorous orthologous relationship in the silkworm and other surveyed insects, indicating conserved functions of these genes during insect evolution. Furthermore, several silkworm BTB protein genes exhibited sex-specific expression in larval tissues or at different stages during metamorphosis. These findings not only contribute to a better understanding of the evolution of insect BTB protein gene families but also provide a basis for further investigation of the functions of BTB protein genes in the silkworm.

  20. Genome-wide analysis of the ATP-binding cassette (ABC) transporter gene family in the silkworm, Bombyx mori.

    Science.gov (United States)

    Xie, Xiaodong; Cheng, Tingcai; Wang, Genhong; Duan, Jun; Niu, Weihuan; Xia, Qingyou

    2012-07-01

    The ATP-binding cassette (ABC) superfamily is a larger protein family with diverse physiological functions in all kingdoms of life. We identified 53 ABC transporters in the silkworm genome, and classified them into eight subfamilies (A-H). Comparative genome analysis revealed that the silkworm has an expanded ABCC subfamily with more members than Drosophila melanogaster, Caenorhabditis elegans, or Homo sapiens. Phylogenetic analysis showed that the ABCE and ABCF genes were highly conserved in the silkworm, indicating possible involvement in fundamental biological processes. Five multidrug resistance-related genes in the ABCB subfamily and two multidrug resistance-associated-related genes in the ABCC subfamily indicated involvement in biochemical defense. Genetic variation analysis revealed four ABC genes that might be evolving under positive selection. Moreover, the silkworm ABCC4 gene might be important for silkworm domestication. Microarray analysis showed that the silkworm ABC genes had distinct expression patterns in different tissues on day 3 of the fifth instar. These results might provide new insights for further functional studies on the ABC genes in the silkworm genome. PMID:22311044

  1. EFFECT OF AQUEOUS MACERATIVES OF SEED POWDER OF SYZIGIUM CUMINI (L ON THE PROTEIN CONTENTS AND MID GUT ENZYME ACTIVITIES IN THE FIFTH INSTAR LARVAE OF SILK WORM, BOMBYX MORI (L (RACE: PM X CSR2

    Directory of Open Access Journals (Sweden)

    Tejas C Korade, Manali T Patil, Kavita H Nimbalkar

    2013-03-01

    Full Text Available Different concentrations (5.0 ppm; 10.0 ppm; 20.0 ppm & 50.0 ppm of the aqueous solution of seed powder of Syzigium cumini (L concentrations was used to treat the leaves of mulberry & fed to the fifth instar larvae of polyvoltine, crossbreed silkworm, Bombyx mori (L (Race : PM x CSR2 for first three days; second day & third day; third day (only. The larvae fed with untreated & water treated leaves were also maintained. Bioassays of proteins (S.P. & T.P. & enzymes (protease & amylase were carried out on fifth day through the use of mid gut homogenate. Treating the mulberry leaves with herbal preparations (Syzigium cumini & feeding them to fifth instar larvae was found reflected into significant improvement in the levels of proteins (S.P. & T.P. & velocities of biochemical reactions catalyzed by protease & amylase. The pattern of increase in soluble proteins & total proteins in the mid gut tissue were 32.147 to 90.074 percent & 5.657 to 39.052 percent respectively. The activities of mid gut protease & amylase were increased by 21.444 to 83.706 percent and 14.54 to 52.257 percent respectively. The nutrient contents of seed powder of Syzigium cumini (L serve to improve the digestibility & exert the influence of efficient metabolism in the fifth instar larvae of silkworm, Bombyx mori (L. The Syzigium seed powder treatment may gear overall biochemical constituency of silkworm larvae, through the significant improvement in the velocity of mid gut enzyme catalyzed biochemical reactions.

  2. Identification and expression profiling of regulatory molecules involved in immune homeostasis in the silkworm, Bombyx mori%家蚕免疫稳态调控分子的鉴定和表达模式分析

    Institute of Scientific and Technical Information of China (English)

    王菲; 李亚明; 化晓婷; 夏庆友

    2012-01-01

    maintenance Of insect immune homeostasis requires prompt activation and down-modulation of the key transcriptional factors: Dorsa/Dif in Toll signaling pathway or Relish in IMD signaling pathway. Several regulatory molecules which modulate the stability or activity of the transcriptional factors in immune response have been identified in Drosophila and some other insects- Mutation or silencing of these molecules leads to over activation of immune system. So far there is no report about the regulatory molecules limiting immune signaling in the silkworm, Bombyx mori. In this study, several molecules which are predicted to be involved in immune homeostasis, including Wnt family members, Ubc9, FAF and POSH, were identified in the silkworm genome by comparative genomic analysis. The expression patterns of these molecules in multiple tissues after microbial infection were recorded, and the results showed that their expression levels generally decreased after microbial infection. Although an increase of more than 1. 5-folds in expression level was observed in certain tissues, a rapid decrease followed and the high level was not maintained. Interestingly, the correspondence between the expression patterns of these molecules and the particular signaling pathway that microbes induced varied in different tissues. This is the first report of the regulatory molecules involved in silkworm immune homeostasis, which provides reference for further investigation on the molecular mechanism of immuno-negative modulation in the ilkworm.%昆虫免疫稳态的维持有赖于准确地激活和有效地抑制Toll或IMD信号通路中的关键转录因子——Dorsal/Dif或Relish.在果蝇等昆虫中,已报道了多种降低转录因子稳定性和活性的免疫稳态调控分子,突变或敲除这类分子导致免疫系统的过度激活.对家蚕Bombyx mori免疫信号通路的研究中,至今为止尚无对这类分子的探索.本研究通过比较基因组学,在家蚕基因组中

  3. Microscopic structural analysis of fractured silk fibers from Bombyx mori and Samia cynthia ricini using 13C CP/MAS NMR with a 1 mm microcoil MAS NMR probehead

    KAUST Repository

    Yamauchi, Kazuo

    2010-07-01

    Conformational changes have been studied in silk fibers from the domestic silkworm Bombyx mori and a wild silkworm Samia cynthia ricini as a result of fractured by stretching. About 300 samples consisting of only the fractured regions of [1-13C]Ala or [1-13C]Gly labeled silk fibers were collected and observed by 13C CP/MAS NMR spectra. The total amount of these fractured fibers is only about 1 mg and therefore we used a home-built 1 mm microcoil MAS NMR probehead. A very small increase in the fraction of random coil was noted for the alanine regions of both silk fibroins and for the glycine region of B. mori silk fibroin. However, there is no difference in the spectra before and after fractured for the glycine region of S. c. ricini silk fibroin. Thus, the influence of fracture occurs exclusively at the Ala region for S. c. ricini. The relationship between sequence, fracture and structure is discussed. © 2010 Elsevier Inc. All rights reserved.

  4. Leaf Surface Scanning Electron Microscopy of 16 Mulberry Genotypes (Morus spp. with Respect to their Feeding Value in Silkworm (Bombyx mori L. Rearing Microscopía Electrónica de Barrido de la Superficie Foliar de 16 Genotipos de Morus spp. en Relación a su Valor Alimenticio para Crianza del Gusano de la Seda (Bombyx mori L.

    Directory of Open Access Journals (Sweden)

    B.K Singhal

    2010-06-01

    Full Text Available Mulberry (Morus spp. is the only silkworm (Bombyx mori L. food plant. In Indian sub tropics, S-146 is the only popular and ruling mulberry genotype for silkworm rearing. As a result, mulberry leaf availability is always the limiting factor, and therefore, sub tropics are contributing less than 1% of the country’s total silk production compared with more than 60% under tropical conditions. Besides climatic conditions, this is due to a very limited number of mulberry genotypes available in this region for silkworm rearing. However, in the mean time, 15 mulberry genotypes viz. ‘Tr-10’,‘Chinese White’,‘K-2’,‘Sujanpur Local’,‘BC2-59’,‘S-1635’,‘C-1730’,‘Mandalaya’,‘S-30’‘(Vishala,‘RFS-175’,‘Anantha’,‘C-2016’,‘C-2017’,‘S-41’ and‘V-1’ were also introduced in the sub tropics, but remained unexplored. In sericulture, leaf surface is also an important parameter for, both, the silkworm’s acceptability of not having any feeding impediment and the mulberry improvement programs. The objective of this study was to explore the possibilities of using these 16 mulberry genotypes for their leaf surface characteristics by scanning electron microscopy and using them for sericulture. Based on leaf yield, stomatal size, stomatal number per unit of area and trichomes and idioblasts length, these genotypes were grouped into different categories. The mulberry genotype ‘Mandalaya’, in addition to the ruling genotype ‘S-146’ excelled because of their higher leaf yield and desired leaf surface characteristics. Furthermore, the genotypes ‘K-2’, ‘S-41’ and ‘Sujanpur Local’ are also suggested to develop high yield mulberry genotypes in the Indian sub tropics.La morera (Morus spp. es la única planta de alimento para el gusano de la seda (Bombyx mori L.. En los sub-trópicos de la India, ‘S-146’ es el único genotipo popular y predominante de morera para criarlo. Como resultado, la

  5. 火麻仁油及甾醇对家蚕寿命影响的观察%Observation of Influence of Semen Cannabis Oil and Sterol on Bombyx Mori Linnaeus Life-span

    Institute of Scientific and Technical Information of China (English)

    李寒冰; 孙静雅; 马永洁; 李根林; 任慧玲

    2012-01-01

    目的:观察火麻仁油及火麻仁甾醇对家蚕寿命的影响,探讨火麻仁油及甾醇的抗衰老作用.方法:取龄期相同、生长整齐的三眠蚕随机分为8组,即空白对照组,火麻仁油高、中、低剂量组,火麻仁甾醇高、中、低剂量组,维生素E对照组,每组110条.使用涂抹法将药液涂抹于新鲜桑叶上,4龄期内喂食3次,5龄期内喂食6次.结果:火麻仁油及甾醇高、中、低剂量组均对家蚕幼虫生存期较空白对照组明显延长(P<0.01或P<0.05),其中火麻仁甾醇能显著延长家蚕全生存期(P<0.01或P<0.05).与空白组相比,火麻仁油及甾醇均能显著延长5龄期家蚕耐饥饿时间(P<0.01).结论:火麻仁油及甾醇均能够延长家香各龄期生存期时间.%Objective:To observe the influence of Semen Cannabis oil and sterol on bombyx mori linnaeus life-span and discuss their anti-aging effect. Methods: Sanmian silkworm with the same age and growth were randomly divided into eight groups, namely blank control group, high, middle, low dose of Semen Cannabis oil group, high, meddle, low dose of Semen Cannabis oil and sterol group, positive control group. Physic liquor is daubed on the mulberry leaf ,3 times during 4 instars and 6 times during 5 instars. Results:The life-span of bombyx mori linnaeus larva was significantly longer in high, meddle, low dose of Semen Cannabis oil and sterol group compared with control group(P<0.01 or P<0.05). Semen Cannabis oil and sterol can significantly extend the all life span(P<0.01 or P<0.05). Compared with the blank control group, Semen Cannabis oil and sterol can significantly extend the hunger resistance time during 5 instars (P <0.01), Conclusion:both Semen Cannabis oil and sterol can extend the life span time of bombyx mori Linnaeus during any instars.

  6. Cloning and sequence analysis of para sodium channel cDNA fragment from silkworm, Bombyx mori%家蚕Para钠通道cDNA片段克隆与序列分析

    Institute of Scientific and Technical Information of China (English)

    何琳; 刘丽花; 汪洋

    2011-01-01

    Previous studies have revealed that a point mutation of a target gene is related to insecticide resistance to pyrethroids. The para sodium channel in the insect central nervous system is the target of pyrethroid insecticides. We used the RT-PCR method to clone the para sodium ion channel in the silkworm, Bombyx mori L. (GenBank No. EF521818).The full length of this cDNA fragment is 4 882 base pairs and its partial ORF is 3 986 bp translated into 1 328 amino acids. BLAST analysis demonstrated that the cloned cDNA fragment is virtually identical to the para sodium channel a subunit gene amplified from other insects. Amino acid homology of the cloned fragment with para sodium channel a subunit genes from Heliothis virescens Fabricius, Aedes aegypti L. , Blattella germanica L. , Drosophila melanogaster Meigen and Musca domestica L. was 95%, 82%, 80%, 79% and 77% respectively.%昆虫神经系统para型钠离子通道是拟除虫菊酯类杀虫剂的主要靶标,已有的研究表明钠离子通道基因发生点突变与昆虫对菊酯类杀虫剂的抗性密切相关.本文通过RT-PCR方法克隆获得了编码家蚕Bombyx mori L.钠离子通道的cDNA片段(GenBank No.EF521818),该片段全长4 882 bp,部分ORF包含3 986 bp核苷酸,翻译成1 328个氨基酸.蛋白序列分析表明,PCR扩增获得的家蚕钠离子通道eDNA片段所编码的氨基酸与其他昆虫的para型钠离子通道α亚基的氨基酸具有很高的同源相似性,与棉铃虫Heliothis virescens Fabricius、埃及伊蚊Aedes aegypti L.、德国小蠊Blattella germanica L.、果蝇Drosophila melanogaster Meigen和家蝇Musca domestica L.的相似性分别为95%、82%、80%、79%、77%.

  7. Efficient silkworm expression of single-chain variable fragment antibody against ginsenoside Re using Bombyx mori nucleopolyhedrovirus bacmid DNA system and its application in enzyme-linked immunosorbent assay for quality control of total ginsenosides.

    Science.gov (United States)

    Sakamoto, Seiichi; Pongkitwitoon, Benyakan; Nakamura, Seiko; Maenaka, Katsumi; Tanaka, Hiroyuki; Morimoto, Satoshi

    2010-09-01

    A single-chain variable fragment (scFv) antibody against ginsenoside Re (G-Re) have been successfully expressed in the silkworm larvae using Bombyx mori nucleopolyhedrovirus (BmNPV) bacmid DNA system. The baculovirus donor vector for expression of scFv against G-Re (GRe-scFv) was constructed to contain honeybee melittin signal sequence to accelerate secretion of the recombinant GRe-scFv into the haemolymph of silkworm larvae. Functional recombinant GRe-scFv was purified by cation exchange chromatography followed by immobilized metal ion affinity chromatography. The yield of purified GRe-scFv was 6.5 mg per 13 silkworm larvae, which is equivalent to 650 mg/l of the haemolymph, exhibiting extremely higher yield than that expressed in Escherichia coli (1.7 mg/l of culture medium). It was revealed from characterization that GRe-scFv retained similar characteristic of the parental monoclonal antibody (MAb) against G-Re (MAb-4G10), making it possible to develop indirect competitive enzyme-linked immunosorbent assay (icELISA) for quality control of total ginsenosides in various ginsengs. The detectable range for calibration of G-Re by developed icELISA shows 0.05-10 microg/ml. These results clearly suggested that the silkworm expression system is quite useful for the expression of functional scFv that frequently required time- and cost-consuming re-folding when it expressed in E. coli. PMID:20592135

  8. LIM-homeodomain transcription factor Awh is a key component activating all three fibroin genes, fibH, fibL and fhx, in the silk gland of the silkworm, Bombyx mori.

    Science.gov (United States)

    Kimoto, Mai; Tsubota, Takuya; Uchino, Keiro; Sezutsu, Hideki; Takiya, Shigeharu

    2015-01-01

    In the silkworm Bombyx mori, three fibroin genes, fibroin-heavy-chain (fibH), fibroin-light-chain (fibL) and fibrohexamerin (fhx), are coexpressed only in the posterior silk gland (PSG) cells, while the sericin genes encoding silk glue proteins are expressed in the middle silk gland (MSG) cells. Silk gland factor-2 (SGF-2) is a PSG-specific activator complex of fibH, composed of a LIM-homeodomain protein, Awh, and its cofactors, Ldb and Lcaf. We investigated whether SGF-2 can activate other fibroin genes using transgenic silkworms. The genes for Ldb and Lcaf were expressed ubiquitously in various tissues, while the gene for Awh was expressed strictly specific in PSG of the wild type silkworms. Misexpression of Awh in transgenic silkworms induced ectopic expression of fibL and fhx as well as fibH in MSG. Coincidently with the induction of fibL and fhx by Awh, binding of SGF-2 to the promoter of fibL and fhx was detected in vitro, and SGF-2 binds directly to the fhx core promoter. Ectopic expression of the fibroin genes was observed at high levels in the middle part of MSG. Moreover, fibL and fhx were induced in the anterior silk gland (ASG) of the transgenic silkworms, but fibH was not. These results indicate that Awh is a key activator of all three fibroin genes, and the activity is probably regulated in conjunction with additional factors. PMID:25449130

  9. Changes in diapause related gene expression pattern during early embryonic development in HCl-treated eggs of bivoltine silkworm Bombyx mori (Lepidoptera: Bombycidae

    Directory of Open Access Journals (Sweden)

    Sirigineedi Sasibhushan

    2013-02-01

    Full Text Available Investigation of differential expression of diapause related genes (five metabolic, five heat shock protein and one translational regulatory in HCl-treated (non-diapause and untreated (diapause eggs of B. mori during early embryogenesis (up to 48h following oviposition revealed the up-regulation of sorbitol dehydrogenase upon HCl treatment, indicating increased glycogen synthesis for further embryonic development but, down-regulation of phosphofructo kinase gene expression after 18h of oviposition indicating an arrest of glycerol and sorbitol conversion. The expression of poly A binding protein gene expression was higher upon HCl treatment, revealing the initiation of translation. The expression levels of other genes analyzed did not vary significantly, except for Hsp90 and Hsp40, which were up-regulated on acid treatment until 18h. Thus, Sorbitoldehydrogenase and phosphofructo kinasegenes have a crucial role in diapause termination as evidenced by HCl treatment, while the other genes did not have major roles.

  10. Evaluation of Impact of Pollen Grains from Bt, Bt/CpTI Transgenic Cotton and Bt Corn Plants on the Growth and Development of the Mulberry Silkworm, Bombyx mori Linnaeus (Lepidoptera: Bombycidae)

    Institute of Scientific and Technical Information of China (English)

    LI Wen-dong; YE Gong-yin; WU Kong-ming; WANG Xiao-qi; GUO Yu-yuan

    2002-01-01

    The δ-endotoxin genes of Bacillus thuringiensis (Bt) and proteinase inhibitor (PI) genes aretwo kinds of genes popularly used for developing transgenic plants resistant to insect pests. To clarify whetherthere is any risk concerning the effects of pollens from these transgenic crops on non-target insects with eco-nomic importance, such as the effects on the growth and development as well as cocoon quality of the silk-worm, Bombyx mori Linnaeus, a series of feeding experiments were conducted, using pollens from transgeniccotton or corn containing crylAc, cry1A+-CpTI or crylAb genes, compared with pollens from non-transgenicnormal cotton and corn as well as the non-pollen treatment. In contrast to the latter ones, pollens from trans-genic plants showed no significant adverse effects on larval mortality, cocoon weight, pupa weight, cocoonshell weight, pupation rate, emergence rate and fecundity of the silkworm after neonates were fed with thepollens for 72 h. In addition, no dosage effects of pollens were found. Though the duration of 1st instar larvaewas prolonged in the case of feeding with transgenic pollens as compared with those of the non-pollen treat-ment, but they were not significantly different from those fed with pollens from non-transgenic cotton or corn.Meanwhile, the body weight of the 3rd instar molters fed with transgenic pollens was obviously different fromthose for non-pollen treatment, and was all significantly heavier than that of the controls. Consequently, it isconsidered that the adverse effect of pollens from transgenic insect-resistant cotton and corn on the growth anddevelopment of the silkworm is negligible.

  11. Genetic Analysis of Izoenzymes Polymorphisms in Silkworm (Bombyx mori L. Strains - doi: 10.4025/actascibiolsci.v35i2.13102

    Directory of Open Access Journals (Sweden)

    Maria Aparecida Fernandez

    2013-05-01

    Full Text Available This work carried out to evaluates the polymorhism in the silkworm of different lineages using the isoenzymes electrophoresis to detect biochemical markers and to investigate the genetics of populations for those lineages. They were used as samples individual extracts of silk glands of second day old larvas of the fifth instar, originating from seven Japanese lineages and eight pure Chinese lineages maintained by the Cocamar-Cooperativa Agroindustrial de Maringá. The isozymes acid phosphatase (ACP, alkaline phosphatase (AKP and carbonic anhydrase (CA they were submitted to the electrophoresis in starch gels 14%. The esterases (EST were analyzed in polyacrylamide gels to 10% and stained with α and b-naphtyl acetate. The total of 21 loci was detected, and 04 (19.05% they are polymorphic, Est-11, Acp, Akp, Ca. The fixation index (Fis for the analyzed isozymes it was 0.0751, indicating excess of homozygotes. The value of Fst (0.6128 it shows that the lineages are well differentiated. The dendrogram obtained with the values of genetic distance didn't separate the Chinese and Japanese lineages analyzed totally. That preliminary evaluation of the lineages of B. mori shows that they present genetic material that it can be used in breeding programs that have the purpose of producing hybrid for silk production.

  12. 新烟碱类杀虫剂对家蚕的急性毒性评价与中毒症状观察%Acute Toxicity Evaluation of Neonicotinoid Insecticides to Bombyx mori and Observation of Toxic Symptoms

    Institute of Scientific and Technical Information of China (English)

    崔新倩; 张骞; 姜辉; 林荣华; 王开运

    2012-01-01

    为明确新烟碱类杀虫剂对非靶标生物家蚕的毒性以及对生态环境的安全性影响,采用浸叶法测定6种新烟碱类杀虫剂及其它3类对照杀虫剂对家蚕的急性毒性,并观察不同种类杀虫剂引起家蚕的急性中毒症状差异.6种新烟碱类杀虫剂中噻虫胺、吡虫啉、噻虫啉和烯啶虫胺对家蚕2龄幼虫96 h的LC50分别为0.065 1、0.174、0.258、0.445 mg/L,属剧毒级农药,噻虫嗪和啶虫脒对家蚕2龄幼虫96 h的LC50分别为1.31、2.73 mg/L,属高毒级农药,6种药剂均对家蚕存在极大的安全风险性.新烟碱类杀虫剂引起家蚕中毒的症状主要表现为拒食,身体扭曲呈“C”或“S”形,头部肿大等.其它3类杀虫剂中,抗生素类杀虫剂阿维菌素的毒性属剧毒级,并在测定药剂中的毒性最高,家蚕中毒症状主要表现为吐液、头部或尾部翘起、拒食等;有机磷类杀虫剂毒死蜱的毒性属高毒级,家蚕中毒症状与新烟碱类杀虫剂相似;吡咯类杀虫剂虫螨腈的毒性属中毒级.因新烟碱类杀虫剂对家蚕的毒性强,建议远离桑园使用,以避免对养蚕生产造成危害.%In order to identify the toxicity of neonicotinoid insecticides to non-target organism silkworm (Bombyx mori) and their impact on the security of ecological environment,we determined and compared the acute toxicity of 6 neonicotinoid insecticides and 3 other insecticides to silkworm with leaf dipping method.The differences in symptoms of acute toxicity caused by treatment with various kinds of insecticides were observed and recorded.The results indicated that,among the 6 neonicotinoid insecticides,LC50 of clothianidin,imidacloprid,thiacloprid and nitenpyram to the 2nd instar silkworm larvae at 96 h was 0.065 1,0.174,0.258 and 0.445 mg/L respectively,being pesticides of virulent toxicity grade.That of thiamethoxam and acetamiprid was 1.31 and 2.73 mg/L respectively,being pesticides of high toxicity grade.They all had

  13. Toxicity evaluation of chlorpyrifos and dimethoate to silkworm variety (Bombyx mori L. 9·Fu) in south China%毒死蜱和乐果对华南地区家蚕品种“9∙芙”的毒性评价

    Institute of Scientific and Technical Information of China (English)

    梅承芳; 张宝兰; 梁燕珍; 张宏涛; 曾国驱; 许玫英; 孙国萍

    2013-01-01

      “9·芙”(Bombyx mori L.9·Fu)作为华南地区饲养的主要家蚕种,考察广泛使用的有机磷农药毒死蜱和乐果对于“9·芙”蚕的毒性和安全性具有重要的经济意义和生态学意义。根据环保部《化学农药环境安全评价试验准则》的要求,首次采用浸叶法评价了w=97%毒死蜱和w=80.7%乐果对华南地区家蚕种“9·芙”(二龄起蚕)的急性毒性效应,根据其毒性范围进行分级,并系统观察了2种杀虫剂处理蚕后的中毒症状。结果表明:采用浸叶法测得的毒死蜱和乐果对“9·芙”蚕的96 h半数致死质量浓度(LC50)分别为2.0 mg·L-1和>400 mg·L-1,根据《化学农药环境安全评价试验准则》,毒死蜱和乐果对“9·芙”蚕分属高毒和低毒药剂。“9·芙”蚕对毒死蜱和乐果的敏感性与其他研究中不同地区蚕种的敏感性具有一定的相似性,且对2种杀虫剂的急性中毒症状具有一定的共性,为典型的有机磷农药中毒症状。蚕因毒死蜱中毒后的症状表现得更为迅速。由此可见,“9·芙”蚕可作为我国农药环境安全性评价中的供试蚕种;毒死蜱属于高毒农药,对于“9·芙”蚕可能存在高风险性,在实际施用中需严格控制施药方式和施药时期;乐果虽属低毒农药,但依然可引起“9·芙”蚕出现中毒效应,在实际施用中也需谨慎对待。%As the main silkworm variety in south China, Bombyx mori L. 9·Fu was used as the testing organism to evaluate the toxicities of the widely used organophosphorus pesticides: chlorpyrifos and dimethoate. Firstly, the acute toxicity effects of 97% chlorpyrifos and 80.7% dimethoate on Bombyx mori L. 9·Fu (the second instar larvae) were evaluated by using the leaf dipping method. The grade was compartmentalized according as toxicity scope and the toxic symptoms of silkworms were systematically observed after treatment with these two

  14. 家蚕副肌球蛋白/小副肌球蛋白基因的克隆及进化分析%cDNA Cloning and Genomic Structure of PM/mPM Gene from B.mori

    Institute of Scientific and Technical Information of China (English)

    徐升胜; 李兵; 许西奎; 沈卫德

    2009-01-01

    [Objective] The experiment aimed to clone Paramyosin/mini-Paramyosin (PM/mPM) gene to analyze the relations between it and moving behaviors. [Method] PCR method and RACE technology were used to obtain whole cDNA of PM and some cDNA of mPM of Bombyx mori. By comparing wgs of Bombyx mori, the genomic sequence of PM/mPM of Bombyx mori was obtained, and, their genome structure was determined. [Result] PM/mPM genes consisted of 17 exons and 16 introns. By the use of selective promoters, The gene sequence encoded PM and mPM, while PM and mPM shared the last 6 exons. The cluster analysis between PM of Bombyx mori and PM of other invertebrate animals demonstrated that the relation between Bombyx mori and Bombyx mandarina was closest and the relation between Bombyx mori and Drosophila melanogaster was farthest in Insecta. [Conclusion] There was no point mutation which could influence flight in amino acid sequence of PM of Bombyx mori and Bombyx mandarina, so the difference of flight capacity of Bombyx mori and Bombyx mandarina might be regulated by other mechanism.

  15. Molecular cloning and bioinformatic analysis of biological clock genes Bmcry1 and Bmcry2 in Bombyx mori%家蚕生物钟基因Bmcryl与Bmcry2的克隆及生物信息学分析

    Institute of Scientific and Technical Information of China (English)

    王文栋; 梁辉; 朱晓苏; 陶卉; 徐丽; 司马杨虎; 徐世清

    2011-01-01

    隐花色素基因(cryptochrome gene,Cry)是已确认的主要生物钟基因之一,它广泛分布于细菌和真核生物中.昆虫Cry基因分为Cry1,和Cry2两类,果蝇只有Cry1,蜜蜂等膜翅目昆虫只有Cry2.为了研究鳞翅目模式昆虫家蚕Bombyx mori的昼夜生物钟分子调控机制和昆虫CRY蛋白的进化,本研究克隆了家蚕Bmcry1与Bmcry2基因的全长cDNA序列,长度分别为2 166 bp和2 389 bp(GenBank登录号分别为HM747059和HM747060),拼接了全基因序列(GenBank 登录号分别为HM747057和HM747058).Bmcry1基因具有12个外显子和11个内含子,Bmcry2具有9个外显子,8个内含子.染色体定位表明Bmcry1和8mcry2分别位于第17号和15号染色体.通过同源建模获得了Bmcry1和Bmcry2蛋白的三维结构,其FAD入口大而深,这与CRY不与嘧啶二聚体结合相符;Bmcry1和Bmcry2表面多为负电荷,只在FAD人口位置有正电荷富集.多序列比对、蛋白质基序和功能域分析、聚类分析等结果显示,Bmcry1和Bmcry2分属昆虫的CRY1和CRY2,与柞蚕Antheraea pernyi等鳞翅目昆虫中CRY蛋白的亲缘关系最近.家蚕的两类CRY与其他昆虫CRY相似,也都具有DNA光解酶和FAD结合功能域,但保守位点和蛋白基序位点不同.本实验为进一步研究家蚕CRY1和CRY2的分子进化机制和功能创造了条件.%Cryptochrome gene (Cry) is one of the major biological clock genes which were widely distributed in bacteria and eukaryotes. Cry genes of insect species are clearly divided into two types, Cry1 and Cry2. Only Cry1 is expressed in Drosophila, while only Cry2 was expressed in bees and other hymenopteran insects. In order to explore the molecular mechanism of circadian clock in lepidopteran model insect Bombyx mori and the evolution of CRY proteins in insect species, we cloned the eDNA sequences of Bmcryl (2 166 bp, GenBank accession no. HM747059) and Bmcry2 (2 389 bp, GenBank accession no.HM747060), and obtained their gene sequences (Gen

  16. Silkworm(Bombyx mori)BmLid is a histone lysine demethylase with a broader specificity than its homolog in Drosophila and mammals

    Institute of Scientific and Technical Information of China (English)

    Bo Zhou; Xiaonan Yang; Jianhao Jiang; Yubing Wang; Minghui Li; Muwang Li; Xuexia Miao; Yongping Huang

    2010-01-01

    @@ Dear Editor, Histone methylation is a dynamic process that plays important roles in gene transcription regulation,and a number of enzymes have been shown to catalyze the removal of methyl marks[1].Shi et al.(2004)identified one of the amino oxidases,lysine-specific demetbylase 1(LSD1),as the first specific demethylase for both mono(me)and dimethylation(me2)of H3K4 and H3K9 in humans[2].Subsequently,a total of 27 JmjC-domaincontaining proteins have been discovered within the human genome,and 15 of them exhibit demethylation activities for specific lysines in the H3 tail[1].

  17. Identification of MBF2 family genes in Bombyx mori and their expression in different tissues and stages and in response to Bacillus bombysepticus infection and starvation.

    Science.gov (United States)

    Zhou, Chun-Yan; Zha, Xing-Fu; Liu, Chun; Han, Min-Jin; Zhang, Li-Ying; Shi, Pan-Pan; Wang, He; Zheng, Ren-Wen; Xia, Qing-You

    2016-08-01

    The Multiprotein bridge factor 2 (MBF2) gene was first identified as a co-activator involved in BmFTZ-F1-mediated activation of the Fushi tarazu gene. Herein, nine homologous genes of MBF2 gene are identified. Evolutionary analysis showed that this gene family is insect-specific and that the family members are closely related to response to pathogens (REPAT) genes. Tissue distribution analysis revealed that these genes could be expressed in a tissue-specific manner. Developmental profiles analysis showed that the MBF2 gene family members were highly expressed in the different stages. Analysis of the expression patterns of nine MBF2 family genes showed that Bacillus bombysepticus treatment induced the up-regulation of several MBF2 family genes, including MBF2-4, -7, -9, -8. Furthermore, we found the MBF2 family genes were modulated by starvation and the expression of these genes recovered upon re-feeding, except for MBF2-5, -9. These findings suggested roles for these proteins in insect defense against pathogens and nutrient metabolism, which has an important guiding significance for designing pest control strategies. PMID:27121992

  18. Isolation and characterization of a humoral factor that stimulates transcription of the acyl-CoA-binding protein in the pheromone gland of the silkmoth, Bombyx mori.

    Science.gov (United States)

    Ohnishi, Atsushi; Koshino, Hiroyuki; Takahashi, Shunya; Esumi, Yasuaki; Matsumoto, Shogo

    2005-02-11

    Acyl-CoA binding protein (ACBP) is a highly conserved 10-kDa intracellular lipid-binding protein that binds straight-chain (C14-C22) acyl-CoA esters with high affinity and is expressed in a wide variety of species ranging from yeast to mammals. Functionally, ACBP can act as an acyl-CoA carrier or as an acyl-CoA pool maker within the cell. Much work on the biochemical properties regarding the ACBP has been performed using various vertebrate and plant tissues, as well as different types of cells in culture, the regulatory mechanisms underlying ACBP gene expression have remained poorly understood. By exploiting the unique sex pheromone production system in the moth pheromone gland (PG), we report that transcription of a specific ACBP termed pheromone gland ACBP is triggered by a hemolymph-based humoral factor. Following purification and structure elucidation by means of high resolution electrospray ionization mass spectrometry and NMR analyses, in conjunction with stereochemical analyses using acid hydrolysates, the humoral factor was identified to be beta-D-glucosyl-O-L-tyrosine. Examination of the hemolymph titers during development revealed that the amount of beta-D-glucosyl-O-L-tyrosine dramatically rose prior to eclosion and reached a maximum of 5 mg/ml (about 1 mg/pupa) on the day preceding eclosion, which was consistent with the effective dose of beta-D-glucosyl-O-L-tyrosine in stimulating pheromone gland ACBP transcription in vivo. Furthermore, in vitro assays using trimmed PG indicated that beta-D-glucosyl-O-L-tyrosine acts directly on the PG. These results provide the first evidence that transcription of some ACBPs can be triggered by specific humoral factors. PMID:15590686

  19. 一种适于研究家蚕色素代谢的眠蚕体壁组织培养方法与黑色素合成抑制试验%A Tissue Culture Method Using Integument of Molting Larva as Material Suitable for Studying Pigment Metabolism in Bombyx mori and Inhibitory Experiment on Melanin Synthesis

    Institute of Scientific and Technical Information of China (English)

    王计英; 李黎; 李海银; 陈萍; 鲁成

    2012-01-01

    利用家蚕幼虫眠时形成新表皮以及重新合成黑色素分布于新表皮上形成固有体色斑纹等特点,用4龄入眠后的家蚕幼虫体壁组织为材料,观察离体培养体壁的色素代谢变化,探索适宜的家蚕幼虫体壁培养基及培养温度条件,眠蚕体壁的取材方法及取材时间,色素代谢观察的外观标志性状等.实验结果表明:含10%优等胎牛血清的Grace培养基适于家蚕幼虫体壁组织培养,培养温度为26℃;取4龄期入眠11 ~12 h后剥去旧表皮的体壁组织更容易观察色素代谢的外观性状变化;体壁培养24h后,刚毛再生和色素沉积是易于观察的色素代谢变化标志性状.通过对家蚕4龄眠蚕离体培养体壁组织的黑色素合成抑制试验,观察到体壁外观性状变化,证明酪氨酸羟化酶催化酪氨酸转化成多巴是家蚕黑色素合成代谢途径中的一个重要环节.%By utilizing the nature of molting Bombyx mori larva which forms new skin by initiating melanin synthesis again and distributing newly synthesized pigments in the new skin to form inherent body markings, we cultured the integument isolated from the molting 4th instar larvae and observed variation of pigment metabolism in the in vitro cultured integument to explore suitable culture medium and temperature, suitable sampling method and sampling time, and typical signs for observing pigment metabolism in integument of molting Bombyx mori larvae. The experimental results showed that: grace medium with 10% high grade fetal bovine serum was suitable for integument tissue culture of Bombyx mori larvae and the suitable culture temperature was 26 ℃; it was much easier to observe appearance change of pigment metabolism by sampling integument underneath the old skin at 11 ~12 h of molting; after culturing the integument for 24 h, seta regeneration and pigment deposition were typical signs of pigment metabolic changes easy to observe. The inhibitory experiment on

  20. Expression and Subcellular Localization of Bombyx mori BmLHep_59 Protein%家蚕BmLHep_59蛋白的表达和亚细胞定位

    Institute of Scientific and Technical Information of China (English)

    侯振宇; 张耀洲

    2011-01-01

    Hepatocellular carcinoma-associated antigen 59 (Hep_59) is one member of the conserved domain family consisting of 100 amino acid residues found mainly in mammals. These proteins mainly exist in eukaryotic cells and most of them are hypothetical proteins. A new gene was obtained from screening the pupa cDNA library of silkworm ( Bombyx mori). Its encoded protein had a conserved domain homologous to Hep_59 and was thus designated as BmLHep_59 (GenBank accession number DN985181 ). Open reading frame (ORF) of the gene was amplified by PCR and cloned into the prokaryotic expression vector pET-28a(+) through EcoR I/Xho I dual enzyme digestion. The recombinant plasmid was transformed into competent E. coli Rosetta (DE3) cells. After being purified with Ni-NTA chromatography, the recombinant protein BmLHep_59 was used as antigen to immunize New Zealand rabbit for preparing polyclonal antibody.ELISA analysis showed that titer of the antiserum was over 1: 12 000. Western blotting analysis indicated that BmLHep_59 was expressed in epidermis, silk gland, and hemolymph of the 5th instar silkworm larvae, among which the highest expression was seen in hemolymph, suggesting its involvement in the regulation of hematopoiesis in silkworm. This protein was also expressed at pupa stage. Subcellular localization showed that BmLHep_59 distributed in both cytoplasm and nucleus of silkworm Bm5 cells.%肝细胞癌相关抗原-59(hepatocellular carcinoma-associated antigen 59,Hep_59)属于哺乳动物中一类含有约100个氨基酸残基的保守结构域家族,这类蛋白主要存在于真核生物细胞中,大多为假想蛋白.从家蚕蛹cDNA文库中筛选的一个新基因,其编码蛋白含有一个和Hep_59高度同源的保守结构域,将其命名为BmLHep_59(GenBank登录号:DN985181).PCR扩增 该基因的ORF,经EcoRⅠ/XhoⅠ双酶切重组到原核表达载体pET-28a(+),并转入E.coli Rosetta(DE3)感受态细胞中表达.通过镍柱亲和层析得

  1. Targated mutagensis in Bombyx mori Using TALENs

    Czech Academy of Sciences Publication Activity Database

    Takasu, Y.; Tamura, T.; Goldsmith, M.; Žurovec, Michal

    Vol. 1338. New York: Springer New York, 2016 - (Kühn, R.; Wurst, W.; Wefers, B.), s. 127-142 ISBN 978-1-4939-2931-3 Institutional support: RVO:60077344 Keywords : silkworm * pBlue-TAL * engineered nucleases Subject RIV: EB - Genetics ; Molecular Biology

  2. 家蚕中肠上皮细胞增殖和分化的初步研究%A Preliminary Study on Proliferation and Differentiation of Intestinal Epithelial Cells of the Silkworm, Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    陈毅彪; 李婷; 郑春琴; 郝向伟; 韦转琴; 向仲怀; 鲁成; 崔红娟

    2013-01-01

    In this study,we indentified the potential location of intestinal stem cells in the silkworm (Bombyx mori)through analysis of proliferation and differentiation of midgut epithelial cells during different developmental stages of larva to pupa.And we observed the morphological structure and cellular component of silkworm midgut during metamorphosis and development of silkworm larvae by Hematoxylin and Eosin (H&E) staining and 4',6-diamidino-2-phenylindole (DAPI) staining.The results showed that the morphological structure and cellular component of silkworm midgut had remarkable changes in the process of molting and metamorphosis of silkworm larvae.The intestinal wall was thin at full appetite stage of each instar,became thicker before molting,and reached peak value at molting stage.There were three types of cells,namely columnar cells (CC),goblet cells (GC) and regenerative cells (RC),in the midgut epithelium.These three types of cells increased gradually with the advance of larval instar.Among them,the goblet cells increased continuously in all instars and reached peak value at molting stage,while the small cells near basal lamina increased at pre-molting stage.Observation by 5-Bromo-2-deoxyUridine (BrdU) and Phospho-histone H3 (PHH3)immunofluorescence staining revealed that midgut epithelial cells,especially the small cells near basal lamina of midgut epithelium,had the highest proliferation rate at premolting stage of each instar.Meanwhile,BrdU label retention assay disp.layed positive signal of BrdU retention in the midgut epithelium near basal lamina.These results demonstrated rapid proliferation of small cells near basal lamina of midgut epithelium during molting of silkworm larvae,suggesting the existence of potential intestinal stem cells in these small cells.%通过分析家蚕自幼虫期到蛹期发育过程中肠上皮细胞的增殖与分化情况,鉴定家蚕中肠干细胞的潜在定位.采用苏木精和伊红(Hematoxylin and Eosin,H&E)染色与4

  3. Transcriptome sequencing and positive selected genes analysis of Bombyx mandarina.

    Science.gov (United States)

    Cheng, Tingcai; Fu, Bohua; Wu, Yuqian; Long, Renwen; Liu, Chun; Xia, Qingyou

    2015-01-01

    The wild silkworm Bombyx mandarina is widely believed to be an ancestor of the domesticated silkworm, Bombyx mori. Silkworms are often used as a model for studying the mechanism of species domestication. Here, we performed transcriptome sequencing of the wild silkworm using an Illumina HiSeq2000 platform. We produced 100,004,078 high-quality reads and assembled them into 50,773 contigs with an N50 length of 1764 bp and a mean length of 941.62 bp. A total of 33,759 unigenes were identified, with 12,805 annotated in the Nr database, 8273 in the Pfam database, and 9093 in the Swiss-Prot database. Expression profile analysis found significant differential expression of 1308 unigenes between the middle silk gland (MSG) and posterior silk gland (PSG). Three sericin genes (sericin 1, sericin 2, and sericin 3) were expressed specifically in the MSG and three fibroin genes (fibroin-H, fibroin-L, and fibroin/P25) were expressed specifically in the PSG. In addition, 32,297 Single-nucleotide polymorphisms (SNPs) and 361 insertion-deletions (INDELs) were detected. Comparison with the domesticated silkworm p50/Dazao identified 5,295 orthologous genes, among which 400 might have experienced or to be experiencing positive selection by Ka/Ks analysis. These data and analyses presented here provide insights into silkworm domestication and an invaluable resource for wild silkworm genomics research. PMID:25806526

  4. Transcriptome sequencing and positive selected genes analysis of Bombyx mandarina.

    Directory of Open Access Journals (Sweden)

    Tingcai Cheng

    Full Text Available The wild silkworm Bombyx mandarina is widely believed to be an ancestor of the domesticated silkworm, Bombyx mori. Silkworms are often used as a model for studying the mechanism of species domestication. Here, we performed transcriptome sequencing of the wild silkworm using an Illumina HiSeq2000 platform. We produced 100,004,078 high-quality reads and assembled them into 50,773 contigs with an N50 length of 1764 bp and a mean length of 941.62 bp. A total of 33,759 unigenes were identified, with 12,805 annotated in the Nr database, 8273 in the Pfam database, and 9093 in the Swiss-Prot database. Expression profile analysis found significant differential expression of 1308 unigenes between the middle silk gland (MSG and posterior silk gland (PSG. Three sericin genes (sericin 1, sericin 2, and sericin 3 were expressed specifically in the MSG and three fibroin genes (fibroin-H, fibroin-L, and fibroin/P25 were expressed specifically in the PSG. In addition, 32,297 Single-nucleotide polymorphisms (SNPs and 361 insertion-deletions (INDELs were detected. Comparison with the domesticated silkworm p50/Dazao identified 5,295 orthologous genes, among which 400 might have experienced or to be experiencing positive selection by Ka/Ks analysis. These data and analyses presented here provide insights into silkworm domestication and an invaluable resource for wild silkworm genomics research.

  5. 一株家蚕病原性微孢子虫的生物学特性与分子系统发育分析%Biological Characters and Molecular Phylogenetic Analysis of a Microsporidian Isolate Pathogenic to Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    罗梅兰; 黄旭华; 潘志新; 蒋满贵; 汤庆坤; 黄深惠; 安春梅; 胡文娟; 甘丽红

    2012-01-01

    GXM,, a new pathogenic microsporidium, was isolated from the silkworm, Bombyx mori. Optical microscopic observation showed that the spore of GXM, was long oval in shape and (1. 85 ?.15)um x (4.19 ?.18)vim in size. The spores had two nuclei at all developmental phases of life cycle and propagated in binary fission. Its developmental was slow. Its life cycle took about 8 to 10 days. There was positive coagulation reaction between GXM, and the antise-rum of Nosema bombycis (Nb). Transmission electron microscopic observation showed that microsporidian GXM, was binuclear and had 11 ~12 polar filament coils. The tilt angle of polar filament was about 45? These biological characters indicated that microsporidian GXM, had the basic taxonomic features of Nosema. Phylogenetic analysts based on SSU rRNA sequences of GXM, and other microsporidian strains and sequence similarity and genetic distance analyses further confirmed that microsporidian GXM, belonged to the genus of Nosema. The 50% infectious concentration (/C50,) of GXM, to newly-hatched silkworm larvae was 6. 06 x106 mL-1, and the germinative infection rate reached 23. 28%. GXM, was a strain of pathogenic microspofidium which had strong infectivity and threat to Bombyx mori.%从家蚕体内分离得到一株新的病原性微孢子虫,编号为GXM1.光学显微镜下观察GXM1微孢子虫为长卵圆形,大小(1.85±0.15) μm×(4.19±0.18)μm,在生活史的各发育阶段均为双核,以二分裂方式增殖,发育速度缓慢,发育周期约8~10 d.GXM1微孢子虫与家蚕微孢子虫(Nb)的抗血清产生阳性凝聚反应.利用透射电子显微镜观察到GXM1微孢子虫的超微结构具双核,极丝11 ~12圈,极丝倾斜角约45°.以上生物学性状显示GXM1微孢子虫具有微孢子虫属(Nosema)的基本分类特征.依据GXM1微孢子虫与其它昆虫微孢子虫的SSU rRNA基因序列构建的系统发育树,以及序列相似性和遗传距离分析,进一步证实GXM1

  6. Infectivity Variation of Nosema bombycis to Bombyx mori Embryonic Cells Under Different Cryopreservation Treatments%家蚕微孢子虫经不同冻存条件处理后对家蚕胚胎细胞的侵染性变化

    Institute of Scientific and Technical Information of China (English)

    马强; 党晓群; 王营; 陈洁; 刘方燕; 赵丽芳; 曹琛福; 吕建强; 潘国庆

    2014-01-01

    为了解家蚕微孢子虫保存方法对其侵染性的影响及大量获得细胞感染实验材料,将采用不同冻存液及冻存法处理的家蚕微孢子虫分别接种于家蚕胚胎细胞(BmE),调查家蚕微孢子虫对细胞的感染率及感染细胞的传代能力.与液氮直接冻存法处理和高温高压灭菌法处理的微孢子虫相比较,用液氮梯度冻存法处理的微孢子虫的粘附率和感染率均显著或极显著提高.采用液氮梯度冻存法以水作冻存液处理的微孢子虫对宿主细胞的感染率为64%,感染细胞能进行连续传代培养;而以含10%二甲基亚砜(DMSO)的胎牛血清作冻存液处理的微孢子虫虽然对宿主细胞的感染率达到80%,但感染细胞在培养7d后大量破裂死亡.因此,选用以水作冻存液经液氮梯度冻存法处理的家蚕微孢子虫有利于感染BmE细胞并在细胞内增殖,从而获得大量的细胞感染实验材料.%In order to study influence of preservation methods on Nosema bombycis infectivity and to acquire infected cells in large quantity for laboratory use,different cryoprotectants and different cryopreservation methods were employed to preserve Nosema bombycis spores which were then used to inoculate Bombyx mori embryonic cells (BmE).The infection rate of microsporidian spores and subculture generation of infected cells were investigated.Compared to microsporidian spores treated by liquid nitrogen direct cryopreservation and autoclaving,microsporidian spores treated by liquid nitrogen gradient cryopreservation had significantly or extremely significantly higher adherence rate and infection rate.Microsporidian spores treated by liquid nitrogen gradient cryopreservation using water as cryoprotectant had an infection rate of 64% to host cells,and the infected cells could be continuously sub-cultured.Although the infectivity of spores preserved using 10% dimethy sulfoxide (DMSO)-containing fetal bovine serum (FBS) as cryoprotectant

  7. 家蚕传染性软化病病毒(桐乡株)5′端非编码区基因的克隆及序列分析%Cloning and Sequence Analysis of 5′ Non-coding Region of Bombyx mori Infectious Flacherie Virus Isolated in Tongxiang,China

    Institute of Scientific and Technical Information of China (English)

    李明乾; 苘娜娜; 蔡顺风; 陈孝学; 金伟; 鲁兴萌

    2009-01-01

    为了探讨家蚕传染性软化病病毒的复制与翻译机制,利用cDNA 5′末端快速扩增(5′-RACE)技术获得了家蚕传染性软化病病毒桐乡分离株(Bombyx mori infectious flacherie virus,BmIFV-2)的5′端非编码区(non-coding region,NCR).序列比较分析发现:BmIFV-2 的5′-NCR由155个核苷酸组成,A+U含量达60.64%,其中包含1个起始密码子AUG;与坂城分离株(BmIFV-1)的5′-NCR相比,BmIFV-2 的5′-NCR在5′末端缺少1个核苷酸,但核苷酸识别率为100%(即单核苷酸变异率为0),而且这个缺少的核苷酸并不影响其5′-NCR的二级结构.与已经证实具有内部核糖体进入位点(internal ribosome entry site,IRES)活性的Iflavirus属的2种病毒EoPV(Ectropis obliqua picorna-like virus)和VDV-1(Varroa destructor virus 1)的5′-NCR相比,BmIFV 的5′-NCR可能也具有IRES活性.

  8. ELECTROPHORETIC SEPARATION AND COMPARATIVE ANALYSIS OF SILK GLAND PROTEINS FROM BOMBYX AND PHILOSAMIA

    OpenAIRE

    Muzafar A Bhat, Punyavathi and Manjunatha H Boregowda*

    2014-01-01

    A comparative analysis of protein extracted from different regions of silk glands in the Bombyx mori L. and Philosamia ricini Hutt was performed employing single-dimensional-electrophoresis technique. Notably, a protein extracted directly from the lumen of the middle silk gland yielded two discrete protein bands with molecular mass of 325 and 26 kDa representing fibroin heavy (H) and low (L) chains than whole silk gland of B. mori. Contrastingly, such differentiation in protein separation cou...

  9. Differential binding of the Bombyx silk gland-specific factor SGFB to its target DNA sequence drives posterior-cell-restricted expression.

    OpenAIRE

    Horard, B; Julien, E; Nony, P; Garel, A; Couble, P

    1997-01-01

    The gene encoding the silk protein P25 in Bombyx mori is expressed in the posterior silk gland (PSG) cells and repressed in the middle silk gland (MSG) cells. To identify the factors involved in this transcription-dependent spatial restriction, we examined the P25 chromatin in PSG and MSG nuclei by DNase I-aided ligation-mediated PCR and analyzed the expression of various P25-lacZ constructs in biolistically treated silk glands. P25 promoter activation depends on two cis-acting elements. One ...

  10. Expression pattern and binding specificity of chemosensory protein BmCSP4 in the silkworm, Bombyx mori%家蚕化学感受蛋白BmCSP4表达谱及结合特性分析

    Institute of Scientific and Technical Information of China (English)

    邓培渊; 乔惠丽; 李丹丹; 鲁云风; 李生才; 阚云超

    2011-01-01

    化学感受蛋白(chemosensory proteins,CSPs)是昆虫体内存在的一类主要识别和运载非挥发性的气味分子和化学刺激物的可溶性蛋白.本研究运用半定量RT-PCR方法分析了BmCSP4的时空及组织表达谱.结果表明:BmCSP4在家蚕Bombyx mori各发育阶段均表达,但表达量从4龄到蛹期逐渐减少,且在雌成虫头部、胸部和腹部表达量较少.用1-NPN作为荧光探针,测定了15种外源配基与BmCSP4蛋白的结合特性,结果显示:仅芳香醛类和芳香酮类化合物在浓度10 μmol/L能将1-NPN从BmCSP4中替换50%,苯甲醛解离常数为3.20 μmol/L,对甲氧基苯甲醛解离常数为2.24 μmol/L,2-戊基-3-苯丙基-烯醛解离常数为2.88μmol/L,1-苯基-1-丁酮解离常数为2.04 μmol/L,苯乙酮解离常数为2.52 μmol/L.据此推测,BmCSP4在不同的发育阶段执行不同的生理功能,并可能参与对芳香醛、芳香酮类气味识别过程.%Chemosensory proteins ( CSPs) are a class of small soluble proteins in insects, which are supposed to recognize and transport non-volatility odour molecules and chemical stimuli. In this study, we investigated the expression pattern of BmCSP4 using semi-quantitative RT-PCR. The results showed that BmCSP4 was expressed across various developmental stages, with gradual decrease from 4th larva to pupa. The tissue expression profile in males and females at adult stage was further studied, and the results showed that the expression level of BmCSP4 was lower in female head ( with antennae removed) , throax and abdomen than in other female tissues. The binding specificity of BmCSP4 to 15 compounds was tested using N-phenyl-1 -naphthylamine (1 -NPN ) as fluorescent probe, and the results showed that only benzaldehyde, p-methoxy benzaldehy, 2-pentyl-3-phenylpropenoic, 1-phenyl-l-butanol and acetophenone at the concentration of 10 junol/L replaced 1-NPN from BmCSP4 by 50% , with the dissociation constants of 3.20, 2.24, 2.88, 2.04 and 2. 52

  11. Yorkie Facilitates Organ Growth and Metamorphosis in Bombyx

    Science.gov (United States)

    Liu, Shumin; Zhang, Panli; Song, Hong-Sheng; Qi, Hai-Sheng; Wei, Zhao-Jun; Zhang, Guozheng; Zhan, Shuai; Liu, Zhihong; Li, Sheng

    2016-01-01

    The Hippo pathway, which was identified from genetic screens in the fruit fly, Drosophila melanogaster, has a major size-control function in animals. All key components of the Hippo pathway, including the transcriptional coactivator Yorkie that is the most critical substrate and downstream effector of the Hippo kinase cassette, are found in the silkworm, Bombyx mori. As revealed by microarray and quantitative real-time PCR, expression of Hippo pathway genes is particularly enriched in several mitotic tissues, including the ovary, testis, and wing disc. Developmental profiles of Hippo pathway genes are generally similar (with the exception of Yorkie) within each organ, but vary greatly in different tissues showing nearly opposing expression patterns in the wing disc and the posterior silk gland (PSG) on day 2 of the prepupal stage. Importantly, the reduction of Yorkie expression by RNAi downregulated Yorkie target genes in the ovary, decreased egg number, and delayed larval-pupal-adult metamorphosis. In contrast, baculovirus-mediated YorkieCA overexpression upregulated Yorkie target genes in the PSG, increased PSG size, and accelerated larval-pupal metamorphosis. Together the results show that Yorkie potentially facilitates organ growth and metamorphosis, and suggest that the evolutionarily conserved Hippo pathway is critical for size control, particularly for PSG growth, in the silkworm. PMID:27489496

  12. Identification of 2nd chromosome region translocated onto the W chromosome by RFLP with EST-cDNA clones in the Gensei-kouken strains of the mulberry silkworm, Bombyx mori L

    Directory of Open Access Journals (Sweden)

    Sivaramakurup Sreekumar

    2010-01-01

    Full Text Available In silkworms, sex-limited strains are either obtained spontaneously or induced by X-rays or gamma rays. When a fragment of an autosome carrying a dominant allele of those genes responsible for certain characters is translocated onto a W chromosome, the female of the successive generations will express these phenotypic characters and sex discrimination can be facilitated. Gensei-kouken strains are sex-limited strains of silkworms developed by irradiating the pupae with gamma rays, by which a portion of the second chromosome is translocated onto the W chromosome. In these improved strains, the females are yellow-blooded and spin yellow cocoons. By using the EST-cDNA clones mapped on the Z chromosome, we identified the sex according to the polymorphic banding pattern or intensity of the signals. Furthermore, by using the clones on the second chromosome, the region of the second chromosome translocated onto the W chromosome was also defined. In both the A95 and A 96 strains selected for the present study, only the mid-portion of the second chromosome was translocated. The differences in length of the fragments translocated in these strains are discussed.

  13. 家蚕氨肽酶N家族基因在5龄幼虫中肠组织的表达分析%Expression Analysis of Aminopeptidase N Family Genes in Midgut Tissue of the Fifth Instar Bombyx mori Larvae

    Institute of Scientific and Technical Information of China (English)

    王猛; 程晨; 郝碧芳; 徐安英; 沈兴家; 黄金山

    2013-01-01

    Aminopeptidase N (APN) is an enzyme that readily hydrolyzes protein or neutral amino acids at the N-terminal of an oligopeptide.It is mainly distributed in the brush border membrane vesicle of midgut epithelial cells of lepidopterous insects and is the important receptor of Bacillus thuringiensis crystal toxins (Cry).To investigate expression patterns of APN family genes,Real-time PCR was employed to analyze the differential expression of APN family genes in larva migut tissue of different silkworm (Bombyx mori) varieties and the expression level of various family members in larva midgut tissue of the same silkworm variety.The results showed that APN family genes were expressed in larva midgut of all tested silkworm varieties.The expression level of APN genes were significantly different between silkworm varieties having different voltinisms (P <0.05).However,there was little difference between silkworm varieties having the same voltinism (P >0.05).The expression levels of APN2,APN4 and APN5 genes were relatively higher in larva midgut tissue of the same silkworm variety,whereas those of APN1 and APN3 were relatively lower.The obtained results would facilitate further study on functioning mechanism of silkworm APN family genes,and provide theoretical basis for breeding silkworm strains resistant to Bacillus thuringiensis.%氨肽酶N(aminopeptidase N,APN)是一种偏好水解蛋白质或寡肽N端中性氨基酸的酶,在鳞翅目昆虫中主要分布于中肠上皮细胞的刷状缘囊膜上,是苏云金芽孢杆菌(Bacillus thuringiensis,Bt)伴孢晶体(Cry)毒素的重要受体蛋白.为了研究家蚕APN家族基因的表达特征,运用Real-time PCR技术检测分析该家族基因在不同家蚕品种幼虫中肠组织的表达差异以及同一家蚕品种幼虫中肠组织中该家族基因各个成员的表达丰度.在所有供试家蚕品种的幼虫中肠组织均可检测到APN家族基因的表达,但不同化性家蚕品种间APN基因的表

  14. Toxicity and Effect of Sublethal Dosage of Several Common Pesticides to Growth and Development of the Silkworm, Bombyx mori%几种常用杀虫剂对家蚕的毒力及亚致死剂量对家蚕生长发育的影响

    Institute of Scientific and Technical Information of China (English)

    罗雁婕; 谢道燕; 柴建萍; 刘永光; 黄平; 丁伟

    2011-01-01

    采用食下毒叶法测定哒螨灵、炔螨特、溴虫腈、敌敌畏、辛硫磷和吡虫啉6种常用杀虫杀螨剂对家蚕的毒力,并研究炔螨特、溴虫腈、敌敌畏和吡虫啉4种杀虫剂的2种亚致死剂量(LC5、LC15)对家蚕生长发育的影响,为蚕区桑园害虫防治选择对家蚕安全的杀虫剂提供参考.结果表明,6种杀虫剂对家蚕的毒力大小依次为吡虫啉>辛硫磷>敌敌畏>哒螨灵>溴虫腈>炔螨特,其对家蚕96 h的LC50分别为0.256、0.556、8.912、50.89、119.715、351.691 mg/L,毒力皆随着时间的累积而增加;炔螨特、溴虫腈、敌敌畏3种杀虫剂的LC5、LC15 2种亚致死剂量对存活家蚕幼虫的历期、眠蚕体质量、全茧量和茧层量与对照相比无明显差异,且相同药剂不同亚致死剂量间对家蚕的影响差异不显著.%In order to provide reference for choosing safe pesticides in controlling pests in mulberry gardens of silkworm ( Bombyx mori) raising area, the toxicity of 6 common pesticides, namely pyridaben, propargite, chlorfenapyr, DDVP, phoxim and imidacloprid, to silkworm was assayed through feeding the larvae with poisoned leaves, and the effect of two sublethal dosages (LC5and LC15) of 4 pesticides, namely propargite, chlorfenapyr, DDVP and imidacloprid, to silkworm growth and development was investigated. The results indicated that the toxicity of pesticides to the silkworm larvae were in the order of imidacloprid > phoxim > DDVP > pyridaben > chlorfenapyr > propargite. Their LC50 to silkworm at 96 h after treatment was 0.256, 0.556, 8.912, 50.689, 119.715 and 351.691 mg/L, respectively. And the toxicity increased with the prolongation of time. The developmental stages, body weight of molting larvae, cocoon weight and cocoon shell weight of survived larvae from treatments at LC5 and LC15 of propargite, chlorfenapyr and DDVP showed no significant difference compared with the control. Moreover, no significant difference

  15. Genome engineering and parthenocloning in the silkworm, Bombyx mori

    Czech Academy of Sciences Publication Activity Database

    Zabelina, V.; Klymenko, V.; Tamura, T.; Doroshenko, K.; Liang, H.; Sezutsu, H.; Sehnal, František

    2015-01-01

    Roč. 40, č. 3 (2015), s. 645-655. ISSN 0250-5991 R&D Projects: GA MŠk(CZ) EE2.3.30.0032; GA ČR GAP502/10/2382 Institutional support: RVO:60077344 Keywords : genomic cloning * genetic engineering * insect biotechnology Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.064, year: 2014 http://www.ias.ac.in/jbiosci/sep2015/645.pdf

  16. Purification and properties of alkaline phosphatase of silkworm Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    TANG Yunming; CEN Liang; CHU Bo; LI Changchun; XU Min; LUO Ying; LU Cheng

    2006-01-01

    Alkaline phosphatase(AKP),from the succus entericus of silkworm,was purified using 10%-50% ammonium sulfate fractions,ion exchange chromatography Of DEAE-Sepharose,and size exclusion chromatography of Sephacryl S-200.The purification fold was 464 times and specified activity was 3936 U/mg.Optimum pH value of the phosphatase was 10.5,and was stable between pH 7.5 and 11.The optimum temperature of the phosphatase was 40℃ and it was unstable over 50℃.Km value of the phosphatase was 1.25 mmol/L.In a given condition,the phosphatase was selectively modified by PCMB,NBS,PMSE TNBS,SUAN,DTT,BrAc,and IAc,the results indicate that PMSF,SUA,BrAc,IAc,and TNBS could Obviously inhibit the activity of the phosphatase,and the degree of inhibition depended on the concentration of these reagents.There was little effect on the activity of phosphatase after treatment by PMSF,DTT,and NBT.We primarily conclude that mercapto and imidazole are essential for AKP from silkworm.Also,Lys residue and disulfide bands are necessary to protect the catalysis of the AKP.

  17. Drosophila heat shock promoter works in transgenic Bombyx mori

    Czech Academy of Sciences Publication Activity Database

    Uhlířová, Miroslava; Asahina, Masako; Riddiford, L. M.; Jindra, Marek

    Kolymbari : University Kolymbari, 2001. s. 10. [International Workshop on Molecular Biology and Genetics of the Lepidoptera. 00.00.2001, Kolymbari] Institutional research plan: CEZ:AV0Z5007907 Keywords : DNA Subject RIV: EB - Genetics ; Molecular Biology

  18. QTL mapping of economically important traits in Silkworm (Bombyx mori)

    Institute of Scientific and Technical Information of China (English)

    2004-01-01

    A backcrossed population(BC1)was derived from a cross between C1AFLP technique was employed for mapping the QTLs.The QTLs for the whole cocoon weight,cocoon shell weight,ratio of cocoon shell,weight of pupae etc.Were analyzed and 11 QTLs were detected based on the constructed linkage map.Two QTLs for whole cocoon weight were localized on linkage group 6 and 19; three QTLs for cocoon shell weight were localized on linkage group 3,14 and 19; three QTLs for ratio of cocoon shell were localized on the linkage group 2,11and 15,and three QTLs for the weight of pupae were localized on linkage 2,14 and 19.All these have laid an important base for the marker assisted breeding of the silkworm.

  19. CYP306A1, a cytochrome P450 enzyme, is essential for ecdysteroid biosynthesis in the prothoracic glands of Bombyx and Drosophila.

    Science.gov (United States)

    Niwa, Ryusuke; Matsuda, Takahiro; Yoshiyama, Takuji; Namiki, Toshiki; Mita, Kazuei; Fujimoto, Yoshinori; Kataoka, Hiroshi

    2004-08-20

    Ecdysteroids mediate a wide variety of developmental and physiological events in insects. In the postembryonic development of insects, ecdysone is synthesized in the prothoracic gland (PG). Although many studies have revealed the biochemical and physiological properties of the enzymes for ecdysteroid biosynthesis, most of the molecular identities of these enzymes have not been elucidated. Here we describe an uncharacterized cytochrome P450 gene, designated Cyp306a1, that is essential for ecdysteroid biosynthesis in the PGs of the silkworm Bombyx mori and fruit fly Drosophila melanogaster. Using the microarray technique for analyzing gene expression profiles in PG cells during Bombyx development, we identified two PG-specific P450 genes whose temporal expression patterns are correlated with changes in ecdysteroid titer during development. Amino acid sequence analysis showed that one of the Bombyx P450 genes belongs to the CYP306A1 subfamily. The temporal and spatial expression pattern of the Drosophila Cyp306a1 homolog is essentially the same as that of Bombyx Cyp306a1. We also found that Drosophila Cyp306a1 is disrupted in the phantom (phm) mutant, known also as the Halloween mutant. The morphological defects and decreased expression of ecdysone-inducible genes in phm suggest that this mutant cannot produce a high titer of ecdysone. Finally we demonstrate that S2 cells transfected with Cyp306a1 convert ketodiol to ketotriol via carbon 25 hydroxylation. These results strongly suggest that CYP306A1 functions as a carbon 25 hydroxylase and has an essential role in ecdysteroid biosynthesis during insect development. PMID:15197185

  20. Identification of ISSR markers associated with productivity traits in silkworm, Bombyx moni L.

    Science.gov (United States)

    Chatterjee, S N; Mohandas, T P

    2003-06-01

    Bombyx mori L., commonly recognised around the world as the mulberry silkworm, is characterized by a wide variability in yield and developmental traits, which have been proven through conventional genetic analysis to be of polygenic nature. A large number of morpho-biochemical traits and RFLP and RAPD markers are mapped on different linkage groups, but to this point very little attention has been given to unravelling the genetics of yield traits. To address this issue, polymorphic profiles of 147 markers generated with 12 ISSR primers on the genomic DNA of 20 silkworm stocks of diverse yield status were subjected to multiple regression and discriminant function analyses (DFA). This led to the identification of eight markers generated by six primers, which demonstrated high beta-coefficient indices of -0.451 to -0.940. Furthermore, a significant difference between the yield traits for stocks with and without the specific marker could also be established. The inheritance pattern of one marker, L13800bp, identified at the first step of selection of markers through stepwise regression analyses for five yield parameters is discussed in the context of applying multiple regression analysis for establishing association, if not linkage, between a group of DNA markers and a particular yield trait of polygenic nature and using such markers in molecular marker-assisted breeding programs. PMID:12834060

  1. High level transactivation by a modified Bombyx ecdysone receptor in mammalian cells without exogenous retinoid X receptor

    OpenAIRE

    Suhr, Steven T.; Gil, Elad B.; Senut, Marie-Claude; GAGE, FRED H.

    1998-01-01

    Our studies of the Bombyx mori ecdysone receptor (BE) revealed that, unlike the Drosophila melanogaster ecdysone receptor (DE), treatment of BE with the ecdysone agonist tebufenozide stimulated high level transactivation in mammalian cells without adding an exogenous heterodimer partner. Gel mobility shift and transfection assays with both the ultraspiracle gene product (Usp) and retinoid X receptor heterodimer partners indicated that this property of BE stems from significantly augmented het...

  2. Constructing and Analyzing Fusion Promoter of Partial Sericin 1 and Bombyx A3 Cytoplasmic Actin

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Previous report showed that the 209 bp DNA sequence upstream of the sericin 1 transcriptional start site (-586 to -378 bp) is involved in promoting transcription and responsible for the tissue specificity of sericin 1 promoter in silkworm Bombyx mori. In the present study, this 209 bp sequence exhibited enhancive effect by assembling in two different locations of ubiquitous Bombyx A3 cytoplasmic actin promoter. Sf-9 cells were transfected with recombinant plasmids using Cellfectin reagent. Firefly luciferase gene located downstream of fusion promoter was considered as a reporter, whereas the activity of the co-transfected Renilla luciferase gene (pGL2-SV40) provides an internal control. This 209 bp region up-regulates the strength of A3 promoter significantly (P < 0.01) when it enters into A3 promoter with respect to the position in sericin 1 gene promoter. This 209-bp fragment was almost functionless when being located upstream of A3 promoter.

  3. POST INFESTATIONAL CHANGES IN THE NUTRITIVE ELEMENTS OF THRIPS (PSEUDODENDROTHRIPS MORI ATTACKED MULBERRY (MORUS SP. FOLIAGE

    Directory of Open Access Journals (Sweden)

    Mahadeva* and MP Shree

    2014-07-01

    Full Text Available Mulberry, the sole food of silkworm, Bombyx mori L. is prone to various pests and diseases. The injury caused during the feeding behavior of insect pests may alter the physiological activities and leading to imbalances in the nutritive constituents of the host plants. An study has been made to know the level of six macro (nitrogen, phosphorus, potassium, calcium, magnesium and sulphur and seven micro nutrients (zinc, iron, manganese, copper, boron, molybdenum and chloride elements in the thrips (Pseudodendrothrips mori Niwa. attacked leaves of six popularly cultivated mulberry varieties. The results revealed a large variation in the nitrogen, phosphorous, potassium, magnesium, sulphur and molybdenum content. The remaining elements were changed non-significantly. Feeding such leaves may cause a variation in growth and development of the silkworm, which results the low yield and poor quality of silk.

  4. Protein expression profiles and the transcriptional analysis of BmLp-c6 in the 5th instar larvae of the silkworm (Bombyx mori) under starvation stress%饥饿胁迫下家蚕5龄起蚕中的蛋白表达谱及BmLp-c6的转录分析

    Institute of Scientific and Technical Information of China (English)

    李迎春; 钟杨生; 林健荣

    2015-01-01

    [目的]分析家蚕Bombyx mori受饥饿胁迫后的蛋白质谱变化,探索其耐受饥饿的机理.[方法]以家蚕品种932为实验材料,利用双向电泳和质谱技术检测5龄起蚕经过24 h饥饿胁迫的蛋白质谱差异变化,利用荧光定量PCR技术分析BmLp-c6的转录表达.[结果]经比对饥饿蚕和正常取食蚕的血淋巴蛋白谱,饥饿蚕有62个特异蛋白点.蛋白点的等电点在4.22 ~6.98之间,分子量分布在20.81 ~144.69 kDa间.选取只在饥饿时出现的特异蛋白点No.7111进行质谱鉴定,根据其氨基酸序列进行引物设计,获得了目的基因BmLp-c6,经与载体pET-21d(+)连接重组后,成功获得诱导表达.经实时荧光定量PCR分析,当5龄起蚕受到饥饿胁迫影响时,BmLp-c6基因在血淋巴中大量转录表达,但在中肠中的转录表达水平却极低.[结论]家蚕5龄起蚕在饥饿胁迫下,血淋巴中的蛋白质谱发生变化,BmLp-c6会大量转录表达.

  5. The Bombyx ovary-derived cell line endogenously expresses PIWI/PIWI-interacting RNA complexes.

    Science.gov (United States)

    Kawaoka, Shinpei; Hayashi, Nobumitsu; Suzuki, Yutaka; Abe, Hiroaki; Sugano, Sumio; Tomari, Yukihide; Shimada, Toru; Katsuma, Susumu

    2009-07-01

    Genetic studies and large-scale sequencing experiments have revealed that the PIWI subfamily proteins and PIWI-interacting RNAs (piRNAs) play an important role in germ line development and transposon control. Biochemical studies in vitro have greatly contributed to the understanding of small interfering RNA (siRNA) and microRNA (miRNA) pathways. However, in vitro analyses of the piRNA pathway have been thus far quite challenging, because their expression is largely restricted to the germ line. Here we report that Bombyx mori ovary-derived cultured cell line, BmN4, endogenously expresses two PIWI subfamily proteins, silkworm Piwi (Siwi) and Ago3 (BmAgo3), and piRNAs associated with them. Siwi-bound piRNAs have a strong bias for uridine at their 5' end and BmAgo3-bound piRNAs are enriched for adenine at position 10. In addition, Siwi preferentially binds antisense piRNAs, whereas BmAgo3 binds sense piRNAs. Moreover, we identified many pairs in which Siwi-bound antisense and BmAgo3-bound sense piRNAs are overlapped by precisely 10 nt at their 5' ends. These signatures are known to be important for secondary piRNA biogenesis in other organisms. Taken together, BmN4 is a unique cell line in which both primary and secondary steps of piRNA biogenesis pathways are active. This cell line would provide useful tools for analysis of piRNA biogenesis and function. PMID:19460866

  6. Caracterização de oito raças do bicho-da-seda (Bombyx mori L. Characterization of eight silkworm races (Bombyx mori L.

    Directory of Open Access Journals (Sweden)

    Antonio José Porto

    2004-02-01

    Full Text Available O experimento foi conduzido na Estação Experimental de Zootecnia de Gália, do Instituto de Zootecnia, SAA-SP, no ano de 2000. Oito raças de bicho-da-seda, de origem Japonesa e Chinesa foram estudadas (B101, B102, B104, B109, C201, C202, C203, C208 em relação a caracteres biológicos (Ganho de peso total de uma lagarta-GP, Porcentagem de mortalidade-MO, Número de machos-NM, Número de fêmeas-NF, Número de ovos/postura-OP e Porcentagem de eclosão-EC e caracteres de produção de casulo (Peso unitário da glândula sericígena-GS, Peso de 30 cascas séricas-CS, Peso de 30 crisálidas-PC, Teor de seda líquido-TS, Casulos desclassificados-CD, Comprimento do casulo-CC e Largura do casulo-LC. O delineamento experimental foi o inteiramente casualizado, com quatro repetições/raça. Não houve variação entre as raças para GP, MO, NF e OP. A raça B101 apresentou, no geral, um menor NM e uma menor EC. Quanto à produção de casulos, no geral, os melhores resultados foram apresentados pela raça C202, com um bom GS (38% do peso final da lagarta, um dos mais altos CS e TS e valores próximos da média para PC, CD, CC e LC. A raça C201, em relação ao casulo produzido, apresentou os piores resultados.The experiment was developed at Estação Experimental de Zootecnia de Galia - Instituto de Zootecnia, SAA-SP, Gália city, São Paulo, Brazil, on 2000. Eight silkworm races of Japanese and Chinese origin were studied (B101, B102, B104, B109, C201, C202, C203, C208 for biological characters ( Total weight-gain for one caterpillar-GP, Percentage of mortality-MO, Number of male-NM, Number of female-NF, Number of egg/laying-OP and Percentage of eclodibility-EC and for characters of cocoon production (silk gland unitary weight-GS, 30 cocoon shell weight-CS, 30 chrysalis weight-PC, silk net purport -TS, disqualified cocoon-CD, cocoon length-CC and cocoon breadth -LC. It was used a completely randomized design, with four replications/ race. It was not detected variation among races for GP, MO, NF and OP. The B101 race showed a smaller NM and a smaller EC. With regard to cocoon production, the C202 race showed the best result, with good GS (38% of the final weight of the caterpillar, one of highest CS and TS and values next to the average for PC, CD, CC e LC. The C201 race, with regard to cocoon produced, showed the worst results.

  7. Genome-wide regulation of innate immunity by juvenile hormone and 20-hydroxyecdysone in the Bombyx fat body

    Directory of Open Access Journals (Sweden)

    Ling Erjun

    2010-10-01

    Full Text Available Abstract Background Insect innate immunity can be affected by juvenile hormone (JH and 20-hydroxyecdysone (20E, but how innate immunity is developmentally regulated by these two hormones in insects has not yet been elucidated. In the silkworm, Bombyx mori, JH and 20E levels are high during the final larval molt (4 M but absent during the feeding stage of 5th instar (5 F, while JH level is low and 20E level is high during the prepupal stage (PP. Fat body produces humoral response molecules and hence is considered as the major organ involved in innate immunity. Results A genome-wide microarray analysis of Bombyx fat body isolated from 4 M, 5 F and PP uncovered a large number of differentially-expressed genes. Most notably, 6 antimicrobial peptide (AMP genes were up-regulated at 4 M versus PP suggesting that Bombyx innate immunity is developmentally regulated by the two hormones. First, JH treatment dramatically increased AMP mRNA levels and activities. Furthermore, 20E treatment exhibited inhibitory effects on AMP mRNA levels and activities, and RNA interference of the 20E receptor EcR-USP had the opposite effects to 20E treatment. Conclusion Taken together, we demonstrate that JH acts as an immune-activator while 20E inhibits innate immunity in the fat body during Bombyx postembryonic development.

  8. 感染家蚕浓核病毒BmDNV1对家蚕抗性和敏感品种生化参数的影响%Effect of BmDNV1 infection on the biochemical parameters in Densonucleosis type1 resistant and susceptible breeds of silkworm, Bombyx mori L.

    Institute of Scientific and Technical Information of China (English)

    Monir SIRAJ; M. BALAVENKATASUBBAIAH; K. CHANDRASEKHARAN; B. NATARAJU; S. D. SHARMA; T. SELVAKUMAR; P. SUDHAKARA RAO

    2007-01-01

    Among silkworm diseases, viral diseases are the major and most common which damages the cocoon production. Viral flacherie is caused by BmIFV, BmDNV1 and BmDNV2. Resistant and susceptible silkworm breeds against BmDNV1 were identified among the Indian Germ plasm stock. The changes in major organic constituents viz.,total protein, carbohydrate and lipid content during the progressive infection of BmDNV1 in susceptible and resistant breeds were estimated as per the standard methods. As the larval age increases, there was a consistent increase in the level of organic constituents viz., protein and carbohydrates in both control and treated batches, but the level of increase in treated batches were markedly less when compared to the respective control batches. After inoculation with BmDNV1, there was a significant decrease in total protein content in haemolymph and mid gut tissue in susceptible breeds when compared with control batches. In resistant breeds, there was a significant decrease in total protein content up to 4 day post inoculation (PI) but subsequently, the decrease was less compared to the respective controls.There was a slight decrease in total carbohydrate content in haemolymph and mid gut tissue in susceptible and resistant breeds. In case of lipid content, there was a significant increase in haemolymph and mid gut tissues in susceptible breeds and in resistant breeds the changes were not found significant. In susceptible silkworm breeds, the biochemical changes clearly indicated that the BmDNV1 infection depletes the major organic constituents viz., total protein and total carbohydrate contents which are the major sources energy. This depletion in these contents resulted in stunted growth in infected silkworms.%在家蚕疾病中,病毒性疾病是危害茧产品的主要且最普遍的疾病.病毒性蚕软腐病是由BrmIFV,BmDNV1和BmDNV2引起的.对印度种质库(Indian Germ plasm stock)中的BmDNV1有抗性和敏感的家蚕品种进行了鉴

  9. Getting a Full Dose? Reconsidering Sex Chromosome Dosage Compensation in the Silkworm, Bombyx mori

    OpenAIRE

    Walters, James R; Hardcastle, Thomas J.

    2011-01-01

    Dosage compensation—equalizing gene expression levels in response to differences in gene dose or copy number—is classically considered to play a critical role in the evolution of heteromorphic sex chromosomes. As the X and Y diverge through degradation and gene loss on the Y (or the W in female-heterogametic ZW taxa), it is expected that dosage compensation will evolve to correct for sex-specific differences in gene dose. Although this is observed in some organisms, recent genome-wide express...

  10. Structure and expression of the silk adhesive protein Ser2 in Bombyx mori

    Czech Academy of Sciences Publication Activity Database

    Kludkiewicz, Barbara; Takasu, Y.; Fedič, Robert; Tamura, T.; Sehnal, František; Žurovec, Michal

    2009-01-01

    Roč. 39, č. 12 (2009), s. 938-946. ISSN 0965-1748 R&D Projects: GA AV ČR IAA5007402; GA MŠk(CZ) LC06077 Institutional research plan: CEZ:AV0Z50070508 Keywords : silkworm * sericin * cocoon Subject RIV: CE - Biochemistry Impact factor: 3.117, year: 2009

  11. THE MODULATORY ROLE OF ZINC IN THE SILKW, BOMBYX MORI (L

    Directory of Open Access Journals (Sweden)

    K Lakshmi Devi

    2013-01-01

    Full Text Available Examining the modulatory effect of zinc chloride on various aspects of silkworm such as Morphometric traits, Protein profiles in various tissues like silk gland, haemolymph, fat body and muscle of the V instar silk worm larvae and also the Economic parameters of the cocoon. Different concentrations of zinc such as 2µg/ml, 5µg/ml, 10µg/ml and 20µg/ml, were prepared by dissolving Zinc chloride in distilled water. The experimental worms were divided in to four groups and fed with mulberry leaves soaked in these selected doses of zinc chloride and control groups of silkworm’s larvae were fed with normal mulberry leaves four times in a day throughout the 5th instar period. Both control and experimental larvae were sacrificed on selected days viz. 1st, 3rd, 5th and 7th day. Cumulatively, the findings of the present study finally suggest that zinc induced active turnover of all profiles of protein metabolic events in the posterior silk gland, creating the conditions that are highly congenial for growth and silk production.The present investigation is one such attempt in the direction of improving the economic parameters of the silk.

  12. Rhodiola rosea extends lifespan and improves stress tolerance in silkworm, Bombyx mori.

    Science.gov (United States)

    Chen, Cong; Song, Jiangbo; Chen, Min; Li, Zhiquan; Tong, Xiaoling; Hu, Hai; Xiang, Zhonghuai; Lu, Cheng; Dai, Fangyin

    2016-04-01

    The root of Rhodiola rosea is widely used in Traditional Chinese Medicine. The extract from R. rosea is reported to extend the lifespan of yeast, nematode, and fruit fly. However, the molecular mechanism is not fully understood. Here, we tested whether R. rosea extends the lifespan of the silkworm. An aqueous extract of R. rosea significantly prolonged the lifespan of the silkworm, without affecting its daily food intake, body weight, or fecundity, suggesting that R. rosea did not exhibit obvious side effects. Rhodiola rosea extract also enhanced the stress resistance in the silkworm, against heat stress (37 °C) and starvation. The R. rosea extract increased the activity of the major antioxidant enzymes, glutathione S-transferase and catalase, and altered the content of glutathione and malondialdehyde. Rhodiola rosea increased the expression of BmFoxO, which is a downstream regulator of insulin/IGF-1 signaling (IIS) pathway in the silkworm. Our results showed that R. rosea extends lifespan, in which IIS pathway might be involved, and enhances stress resistance in the silkworm. Thus, the silkworm might be used as a novel animal model for lifespan study and efficacy evaluation of Traditional Chinese Medicines. PMID:26497336

  13. Effects of heavy-ion radiosurgery on the hemopoietic function of the silkworm Bombyx mori

    International Nuclear Information System (INIS)

    To study the effects of heavy-ion radiosurgery on the hemopoietic function of a silkworm, hemopoietic organs of larvae were locally irradiated with carbon-ion beams, and the changes in the hemocyte density and in the hemocyte function were investigated. When the larvae were irradiated by 50 Gy to 300 Gy carbon ions on the 3rd day of the 4th instar, the hemocyte densities did not change for a while, though they gradually increased at a later stage, but were finally still significantly lower than those of unirradiated controls. The hemocyte densities of the larvae irradiated at different developmental stages showed suppressed increments, and carbon-ion irradiation given to larvae at early stages compared to the later stages had a significant suppressive effect on the hemocyte densities. On unilateral irradiated larvae a hemocyte intermediate increment between those of bilateral irradiated larvae and unirradiated controls was observed. The percentage of dead hemocytes was obviously higher for irradiated larvae than unirradiated controls during the later 5th instar. Thus, it is evident that carbon-ion radiosurgery on hemopoietic organs of silkworm induced not only a quantitative change, but also a qualitative change in the hemocytes. (author)

  14. The nicotinic acetylcholine receptor gene family of the silkworm, Bombyx mori

    OpenAIRE

    Zhang Chuan-Xi; Dong Ke; Shao Ya-Ming

    2007-01-01

    Abstract Background Nicotinic acetylcholine receptors (nAChRs) mediate fast synaptic cholinergic transmission in the insect central nervous system. The insect nAChR is the molecular target of a class of insecticides, neonicotinoids. Like mammalian nAChRs, insect nAChRs are considered to be made up of five subunits, coded by homologous genes belonging to the same family. The nAChR subunit genes of Drosophila melanogaster, Apis mellifera and Anopheles gambiae have been cloned previously based o...

  15. Two adenine nucleotide translocase paralogues involved in cell proliferation and spermatogenesis in the silkworm Bombyx mori.

    Directory of Open Access Journals (Sweden)

    Ryohei Sugahara

    Full Text Available Mitochondrial adenine nucleotide translocase (ANT specifically acts in ADP/ATP exchange through the mitochondrial inner membrane. This transporter protein thereby plays a significant role in energy metabolism in eukaryotic cells. Most mammals have four paralogous ANT genes (ANT1-4 and utilize these paralogues in different types of cells. The fourth paralogue of ANT (ANT4 is present only in mammals and reptiles and is exclusively expressed in testicular germ cells where it is required for meiotic progression in the spermatocytes. Here, we report that silkworms harbor two ANT paralogues, the homeostatic paralogue (BmANTI1 and the testis-specific paralogue (BmANTI2. The BmANTI2 protein has an N-terminal extension in which the positions of lysine residues in the amino acid sequence are distributed as in human ANT4. An expression analysis showed that BmANTI2 transcripts were restricted to the testis, suggesting the protein has a role in the progression of spermatogenesis. By contrast, BmANTI1 was expressed in all tissues tested, suggesting it has an important role in homeostasis. We also observed that cultured silkworm cells required BmANTI1 for proliferation. The ANTI1 protein of the lepidopteran Plutella xylostella (PxANTI1, but not those of other insect species (or PxANTI2, restored cell proliferation in BmANTI1-knockdown cells suggesting that ANTI1 has similar energy metabolism functions across the Lepidoptera. Our results suggest that BmANTI2 is evolutionarily divergent from BmANTI1 and has developed a specific role in spermatogenesis similar to that of mammalian ANT4.

  16. Biochemical characterization of novel bioactive protein from silkworm (Bombyx mori L) fecal matter.

    Science.gov (United States)

    Raghavendra, R; Neelagund, S E

    2012-07-01

    In this study, complete purification and biochemical characterization of protein is presented. The protein was purified by using Sephadex G-75 gel filtration column followed by reverse-phase high-performance liquid chromatography in a C18 column. The molecular weight of the protein was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis, mass spectrum matrix-assisted laser desorption/ionization-time-of-flight-mass spectrometry (MALDI-TOF-MS) and liquid chromatography-electrospray ionization tandem mass spectrometry. Protein was fragmented by trypsin based on the m/z values obtained by MALDI-TOF-MS analysis. The peptide fragments sequence showed homology with DEAD-box-ATP-dependent RNA helicase 45, present in a public domain, National Centre for Biotechnology Information. The protein exhibited antibacterial activity against selected Gram +/- bacteria. The analgesic activity was determined by conducting acetic-acid-induced writhing test in mice. PMID:22328263

  17. The properties of monoclonal antibody against sepiapterin reductase from fat body of the silkworm, Bombyx mori.

    Science.gov (United States)

    Iino, T; Sawada, H; Gyure, W L; Tsusué, M

    1992-12-01

    A specific monoclonal antibody prepared for the 29-kDa a subunit of silkworm fat body sepiapterin reductase (SPR) was able to recognize the subunit in crude extract of fat body after SDS treatment. Although SPR from the silkworm fat body has biochemical properties similar to those reported for SPR from mammalian sources, especially rat erythrocytes, the antibody failed to recognize the 28-kDa subunit of rat erythrocyte SPR. This result indicates that SPR from silkworm fat body has a different amino-acid sequence from that of the rat erythrocyte enzyme. Sepiapterin reductase activity has not been found in crude extract of fat body from the silkworm mutant lemon. Although the antibody recognized only 29-kDa protein in the crude extract of silkworm fat body from normal strain after SDS-treatment, the antibody recognized only an approximately 80-kDa protein in the crude extract of the lemon mutant after SDS-treatment. PMID:1292509

  18. Production of Recombinant Human DNA Polymerase Delta in a Bombyx mori Bioreactor

    OpenAIRE

    Zhou, Yajing; Chen, Huiqing; Li, Xiao; Wang, Yujue; Chen, Keping; Zhang, Sufang; MENG, XIAO; Lee, Ernest Y. C.; Lee, Marietta Y.W.T.

    2011-01-01

    Eukaryotic DNA polymerase δ (pol δ) plays a crucial role in chromosomal DNA replication and various DNA repair processes. It is thought to consist of p125, p66 (p68), p50 and p12 subunits. However, rigorous isolation of mammalian pol δ from natural sources has usually yielded two-subunit preparations containing only p125 and p50 polypeptides. While recombinant pol δ isolated from infected insect cells have some problems of consistency in the quality of the preparations, and the yields are muc...

  19. Production of recombinant human DNA polymerase delta in a Bombyx mori bioreactor.

    Science.gov (United States)

    Zhou, Yajing; Chen, Huiqing; Li, Xiao; Wang, Yujue; Chen, Keping; Zhang, Sufang; Meng, Xiao; Lee, Ernest Y C; Lee, Marietta Y W T

    2011-01-01

    Eukaryotic DNA polymerase δ (pol δ) plays a crucial role in chromosomal DNA replication and various DNA repair processes. It is thought to consist of p125, p66 (p68), p50 and p12 subunits. However, rigorous isolation of mammalian pol δ from natural sources has usually yielded two-subunit preparations containing only p125 and p50 polypeptides. While recombinant pol δ isolated from infected insect cells have some problems of consistency in the quality of the preparations, and the yields are much lower. To address these deficiencies, we have constructed recombinant BmNPV baculoviruses using MultiBac system. This method makes the generation of recombinant forms of pol δ containing mutations in any one of the subunits or combinations thereof extremely facile. From about 350 infected larvae, we obtained as much as 4 mg of pol δ four-subunit complex. Highly purified enzyme behaved like the one of native form by rigorous characterization and comparison of its activities on poly(dA)/oligo(dT) template-primer and singly primed M13 DNA, and its homogeneity on FPLC gel filtration. In vitro base excision repair (BER) assays showed that pol δ plays a significant role in uracil-intiated BER and is more likely to mediate LP BER, while the trimer lacking p12 is more likely to mediate SN BER. It seems likely that loss of p12 modulates the rate of SN BER and LP BER during the repair process. Thus, this work provides a simple, fast, reliable and economic way for the large-scale production of human DNA polymerase δ with a high activity and purity, setting up a new platform for our further research on the biochemical properties of pol δ, its regulation and the integration of its functions, and how alterations in pol δ function could contribute to the etiology of human cancer or other diseases that can result from loss of genomic stability. PMID:21789240

  20. Biosynthesis and characterization of typical fibroin crystalline polypeptides of silkworm Bombyx mori

    International Nuclear Information System (INIS)

    We aimed to investigate the self-organization/self-assembly mechanisms of silkworm fibroin-based material. In the present study, for the first time, we designed and multimerized four DNA 'monomer' sequences from structurally simple fibroin crystalline peptides or analog, [GAGAGX] (X = A, S, Y and V) to encode polypeptides [GAGAGX]16 (eGA, eGS, eGY and eGV) using a 'head-to-tail' construction strategy. Multimers were cloned into pGEX-KG and fusion proteins GST-[GAGAGX]16 (KGA, KGS, KGY and KGV) were efficiently expressed in Escherichia coli. These fusion proteins were isolated and purified by GST affinity chromatography and confirmed by SDS-PAGE and Western blot analysis using antibody reactive to GST. The polypeptides were cleavaged from GST fusion proteins by digesting with thrombin enzyme. The composition of the four polypeptides was confirmed by composition analysis of amino acids, and their abilities to form β-sheet structure were determined by ThT fluorescence spectral analysis. The content of β-sheet among the four polypeptides followed the order: eGS > eGV > eGY > eGA.

  1. The effect of BmNPV infection on protein metabolism in silkworm (Bombyx mori larva

    Directory of Open Access Journals (Sweden)

    K Etebari

    2007-02-01

    Full Text Available Grasseri is one of the most important diseases of silkworm with significant yield loss, which is caused by nuclear polyhedrosis viruses (NPV. In the present research the effect of this disease on changes of biochemical compounds which are related to protein metabolism in 5th instar larvae were studied. The larvae that showed the grasseri symptoms after contamination with 5.5×10-4 polyhedral/ml were assumed as infected treatment. The hemolymph of infected and uninfected larvae in 3 and 5 days after 4th molting were collected and its total protein, urea, alanine aminotransferase (ALT and aspartate aminotransferase (AST were measured. The results showed that the amount of all the compounds except urea were considerably different in both groups. Total protein had decreased in infected larvae but activity level of two aminotransferases significantly increased. Therefore, grasseri has a considerable effect on protein metabolism.

  2. The effect of BmNPV infection on protein metabolism in silkworm (Bombyx mori) larva

    OpenAIRE

    K Etebari; L Matindoost; SZ Mirhoseini; MW Turnbull

    2007-01-01

    Grasseri is one of the most important diseases of silkworm with significant yield loss, which is caused by nuclear polyhedrosis viruses (NPV). In the present research the effect of this disease on changes of biochemical compounds which are related to protein metabolism in 5th instar larvae were studied. The larvae that showed the grasseri symptoms after contamination with 5.5×10-4 polyhedral/ml were assumed as infected treatment. The hemolymph of infected and uninfected larvae in 3 and 5 days...

  3. Biochemical characterization of maintenance DNA methyltransferase DNMT-1 from silkworm, Bombyx mori.

    Science.gov (United States)

    Mitsudome, Takumi; Mon, Hiroaki; Xu, Jian; Li, Zhiqing; Lee, Jae Man; Patil, Anandrao Ashok; Masuda, Atsushi; Iiyama, Kazuhiro; Morokuma, Daisuke; Kusakabe, Takahiro

    2015-03-01

    DNA methylation is an important epigenetic mechanism involved in gene expression of vertebrates and invertebrates. In general, DNA methylation profile is established by de novo DNA methyltransferases (DNMT-3A, -3B) and maintainance DNA methyltransferase (DNMT-1). DNMT-1 has a strong substrate preference for hemimethylated DNA over the unmethylated one. Because the silkworm genome lacks an apparent homologue of de novo DNMT, it is still unclear that how silkworm chromosome establishes and maintains its DNA methylation profile. As the first step to unravel this enigma, we purified recombinant BmDNMT-1 using baculovirus expression system and characterized its DNA-binding and DNA methylation activity. We found that the BmDNMT-1 preferentially methylates hemimethylated DNA despite binding to both unmethylated and hemimethylated DNA. Interestingly, BmDNMT-1 formed a complex with DNA in the presence or absence of methyl group donor, S-Adenosylmethionine (AdoMet) and the AdoMet-dependent complex formation was facilitated by Zn(2+) and Mn(2+). Our results provide clear evidence that BmDNMT-1 retained the function as maintenance DNMT but its sensitivity to metal ions is different from mammalian DNMT-1. PMID:25623240

  4. The microbicidal effect of the silkworm (bombyx mori) hemocytes is not dependent on oxygen reactive matabolites

    Czech Academy of Sciences Publication Activity Database

    Hyršl, P.; Lojek, Antonín; Číž, Milan; Kubala, Lukáš

    Bratislava: VEDA, 2002 - (Timko, J.; Turňa, J.; Pastorek, J.). s. 204 [Biochemický Zjazd /18./. 10.09.2002-13.09.2002, Stará Lesná] Institutional research plan: CEZ:AV0Z5004920 Keywords : hemocytes * reactive oxygen species * silkworn Subject RIV: BO - Biophysics

  5. Impact of botanical extracts on histopathology of silkworm (Bombyx mori L.

    Directory of Open Access Journals (Sweden)

    Mude Jagadish Naik

    2015-06-01

    Full Text Available Present study was conducted to find out the effect of various botanical extract on the tissue, cellular an d sub cellular level and histopathology of silkworm, findings of the present study gives useful data concerning the changes in the insect. Three plants extract viz Azadirachta indica, Ocimum sanctum and Parthenium hysterophorus were used as experimental while untreated leaves consider as control. These botanicals were sprayed on the tukra (Pink mealy bug infected mulberry leaves and feed to silkworm (CSR2 bivoltine hybrid. Findings of the study suggested no change in the fat body of the silkworm feed on the botanical sprayed leaves and it was with normal vacuolization cytoplasm of cells. While hypertrophied nucleus fat body and voculated cytoplasm was reported in the silkworm fed on the tukra infected chawki leaves. The outer layers of the nucleolus were reported somewhat hypertrophied and cytoplasm was reported vacuolate with mild degeneration of cell in silkworm fed on the tukra infected leaves. Silk worm fed leaves revealed almost similar changes to that of normal and there was no change in botanical sprayed fed larvae. The impact in tissue of the silkworm when fed with normal and crude botanical extracts against mealy bugs shows normalcy, but in the t ukra infected mulberry leaves fed by silk worms the tissues sho ws slight degenerative with nutritional impact upon them

  6. The adverse effects of phoxim exposure in the midgut of silkworm, Bombyx mori.

    Science.gov (United States)

    Gu, ZhiYa; Zhou, YiJun; Xie, Yi; Li, FanChi; Ma, Lie; Sun, ShanShan; Wu, Yu; Wang, BinBin; Wang, JuMei; Hong, Fashui; Shen, WeiDe; Li, Bing

    2014-02-01

    The silkworm is an important economic insect. Poisoning of silkworms by organophosphate pesticides causes tremendous loss to the sericulture. In this study, Solexa sequencing technology was performed to profile the gene expression changes in the midgut of silkworms in response to 24h of phoxim exposure and the impact on detoxification, apoptosis and immune defense were addressed. The results showed that 254 genes displayed at least 2.0-fold changes in expression levels, with 148 genes up-regulated and 106 genes down-regulated. Cytochrome P450 played an important role in detoxification. Histopathology examination and transmission electron microscope revealed swollen mitochondria and disappearance of the cristae of mitochondria, which are the important features in insect apoptotic cells. Cytochrome C release from mitochondria into the cytoplasm was confirmed. In addition, the Toll and immune deficiency (IMD) signal pathways were all inhibited using qRT-PCR. Our results could help better understand the impact of phoxim exposure on silkworm. PMID:23899924

  7. Examples of Mori dream spaces with Picard number two

    OpenAIRE

    Ito, Atsushi

    2013-01-01

    In this note, we give a sufficient condition such that a projective variety with Picard number two is a Mori dream space. Using this condition, we obtain examples of Mori dream spaces with Picard number two.

  8. On images of Mori dream spaces

    CERN Document Server

    Okawa, Shinnosuke

    2011-01-01

    The purpose of this paper is to study the geometry of the images of morphisms from Mori dream spaces. Firstly we prove that a variety which admits a surjective morphism from a Mori dream space again is a MDS. Secondly we introduce a fan structure on the effective cone of a MDS and show that under a surjective morphism between MDSs the fan of the target space coincides with the restriction of the fan of the source. We see that this fan encodes some information of the Zariski decompositions, which in turn is equivalent to the information of the GIT equivalence of VGIT of the Cox ring. Corresponding to these two interpretations, two different proofs are given to the second theorem. Generalizations to non-\\mathbb{Q}-factorial cases and to Mori dream re- gions are also treated.

  9. Neuro Memento Mori: meditations on death

    OpenAIRE

    Prophet, Jane

    2015-01-01

    In Neuro Memento Mori, digital animations and live action video are projection mapped onto a 3D print of the artist’s head and neck made from data from 3D scans of the head and MRI scans of the brain. The life-sized 3D printed sculptures are dissected to reveal the artist’s brain and ‘make real’ fMRI data gathered as the artist conducted experiments in the MRI scanner including viewing memento mori paintings and meditating on death. Computation is used to produce 3D neuroimages, 3D prints and...

  10. A study on interaspecific biodiversity of eight groups of silkworm (Bombyx mori) by biochemical markers

    Institute of Scientific and Technical Information of China (English)

    KAYVANETEBARI; S.Z.MIRHOSEINI; L.MATINDOOST

    2005-01-01

    The recognition of biodiversity in different races and lines of silkworm (Bombyx mori) is very useful for breeding programs and production of high efficiency hybrids. In this study eight groups of silkworm were selected including 103, 107, Xihang 1 and 2 of Japanese origin and 104, 110, Koming 1 and 2 of Chinese origin. The activity levels of three enzymes including alkaline phosphatase, alanine aminotransferase and aspartate aminotransferase in haemolymph of fifth instar larva were measured. Moreover, the quantitative amount of total protein, cholesterol and glucose of haemolymph was evaluated.The data reveal that the activity level of measured macromolecules except for alkaline phosphatase were significantly different in all the groups. Hierarchical agglomerative clustering under UPGMA model separated line 104 from other groups. Two groups of Koming 1 and Xihang 1 had the most intergroup similarities.

  11. Intercultural Usage of Mori Folium: Comparison Review from a Korean Medical Perspective

    OpenAIRE

    Byungjin Joh; Eun Sang Jeon; Su Hye Lim; Yu Lee Park; Wansu Park; Han Chae

    2015-01-01

    Objectives. A review on studies related to the use of Mori folium, the leaves of Morus alba, was conducted with the goal of identifying new clinical applications in Korean medicine. Methods. Global literature search was conducted using three electronic databases up to January 2015 with the term Morus alba and its Korean terms. KM literatures including textbooks and standard pharmacopoeia were separately hand-searched and reviewed to provide comparison. Data were extracted according to predete...

  12. Purification and Functional Characterization of a Protein: Bombyx mori Human Growth Hormone Like Protein in Silkworm Pupa

    OpenAIRE

    Jianqing Chen; Tejun Shu; Zhengbing Lv; Zuoming Nie; Jian Chen; Hao Chen; Wei Yu; Qijing Gai; Yaozhou Zhang

    2014-01-01

    Human growth hormone (hGH) is a peptide hormone secreted by eosinophils of the human anterior pituitary, and a regulatory factor for a variety of metabolic pathways. A 30-kD protein from the pupa stage of silkworm was detected by Western blotting and confirmed by immunoprecipitation based on its ability to bind to anti-hGH antibody. This protein, named BmhGH-like protein, was purified from fresh silkworm pupas through low-temperature homogenization, filtration, and centrifugation to remove la...

  13. Effect of solvents on properties of Bombyx mori silk grafted by methyl methacrylate (MMA and methacrylamide (MAA

    Directory of Open Access Journals (Sweden)

    Wattana Klairatsamee

    2005-11-01

    Full Text Available Mulberry silks were chemically modified in order to increase weight gain, resulting from degumming process using graft copolymerisation technique with vinyl monomers, i.e. MMA, MAA and MMA/MAA. Due to the appearance of PMMA homopolymer granules adhered on the MMA- and MMA/MAA-grafted silk surfaces resulting in surface roughness when silk was grafted by MMA in water, the influence of grafting solvents was examined, using different water/ethanol volume ratios of 100/0, 75/25, 50/50, 25/75 and 0/100. FTIR spectra of the grafted silks presented the absorption bands of the vinyl monomers used for the grafting process. In addition, high values of % polymer add-on were obtained for all of the grafted silks. It was also found that the suitable solvents were 25/75 water/ethanol for the silk grafted by MMA and MMA/MAA, and water for the silk grafted by MAA, in order to get the smooth grafted silk surface and high polymer add-on. Moreover, all the grafted silks showed slightly greater stiffness, as indicated by the increase of Young's modulus and the decrease of elongation.

  14. Extensive conserved synteny of genes between the karyotypes of Manduca sexta and Bombyx mori revealed by BAC-FISH mapping

    Czech Academy of Sciences Publication Activity Database

    Yasukochi, Y.; Tanaka-Okuyama, M.; Shibata, F.; Yoshido, A.; Marec, František; Wu, Ch.; Zhang, H.; Goldsmith, M. R.; Sahara, K.

    2009-01-01

    Roč. 4, č. 10 (2009), e7465. E-ISSN 1932-6203 R&D Projects: GA ČR GA206/06/1860; GA AV ČR IAA600960925 Grant ostatní: Japan Society for the Promotion of Science(JP) 18380037; National Science Foundation(US) IBN020838 Institutional research plan: CEZ:AV0Z50070508 Keywords : Manduca sexta Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.351, year: 2009

  15. Multiple Interval Mapping for Whole Cocoon Weight and Related Economically Important Traits QTL in Silkworm (Bombyx mori)

    Institute of Scientific and Technical Information of China (English)

    LI Bin; LU Cheng; ZHAO Ai-chun; XIANG Zhong-huai

    2006-01-01

    A backcrossed population (BCi) derived from a cross between C100 and Dazao was obtained. The quantitative trait loci (QTLs) of the economically important traits for whole cocoon weight, cocoon shell weight, ratio of cocoon shell and weight of pupae, etc., were analyzed for the first time using the multiple interval mapping software WinQTLCart2.0. In total 40 QTLs were detected and contributed to 21 groups based on the constructed linkage map. According to the mapping results, 2, 2, 3, and 2 major QTLs explained over 20% of total phenotypic variations, whereas four QTLs, namely qCW-19,qSW-2, qCSR-4, and qPW-23, explained more than 30% of total phenotypic variations for whole cocoon weight, cocoon shell weight, ratio of cocoon shell and weight of pupae, respectively. Correlated traits QTLs often share the same location.Furthermore, most of the detected QTLs were closed to one-side marker. By using the very closed markers, positive QTLs can be aggregated, which can form a basis for molecular marker-assisted selection and breeding.

  16. Extensive Conserved Synteny of Genes between the Karyotypes of Manduca sexta and Bombyx mori Revealed by BAC-FISH Mapping

    OpenAIRE

    Yasukochi, Yuji; Tanaka-Okuyama, Makiko; Shibata, Fukashi; Yoshido, Atsuo; Marec, František; Wu, Chengcang; Zhang, Hongbin; Goldsmith, Marian R; Sahara, Ken

    2009-01-01

    Background Genome sequencing projects have been completed for several species representing four highly diverged holometabolous insect orders, Diptera, Hymenoptera, Coleoptera, and Lepidoptera. The striking evolutionary diversity of insects argues a need for efficient methods to apply genome information from such models to genetically uncharacterized species. Constructing conserved synteny maps plays a crucial role in this task. Here, we demonstrate the use of fluorescence in situ hybridizatio...

  17. In vitro conditions for 14C-leucine incorporation into the protein of cultured ovaries of the silkworm, Bombyx mori

    International Nuclear Information System (INIS)

    Vitellogenic ovaries of silkworm pupae were incubated in vitro in different media based on the Wyatt's medium to establish an adequate condition for culture of silkworm ovaries. Incorporation of 14C-leucine into protein fraction was determined to assess the biochemical activity of the ovary. When ovaries were incubated in vitro for a short time by 6 hr, a saturation kinetics of incorporation of the labelled leucine was shown. Sequential substitution of K+ ion to Na+ ion in the medium had no effect on the incorporation of 14C-leucine, but Mg2+ ion appeared to stimulate synthetic activity at more than 10 mM. The activity was not affected at pH range 5.0-7.2. Neither different sugars, nor vitellogenin nor lipoprotein prepared from silkworm haemolymph affected the incorporation of 14C-leucine, when added into the medium. The synthesis of protein depended upon the developmental stages of the cultured ovaries and was most active in 6-day-old ovary. Ovaries developing in pupal body showed comparable changes in synthetic activity. It is concluded that the chemical composition of the medium does not exert a strict effect on synthetic activity of protein in short-term cultures and the ovaries cultured in vitro maintain the activity comparable with those found in in situ condition. (author)

  18. Crystal structure of Bombyx mori arylphorins reveals a 3:3 heterohexamer with multiple papain cleavage sites

    OpenAIRE

    Hou, Yong; Li, Jianwei; Li, Yi; Dong, Zhaoming; Xia, Qingyou; Yuan, Y. Adam

    2014-01-01

    In holometabolous insects, the accumulation and utilization of storage proteins (SPs), including arylphorins and methionine-rich proteins, are critical for the insect metamorphosis. SPs function as amino acids reserves, which are synthesized in fat body, secreted into the larval hemolymph and taken up by fat body shortly before pupation. However, the detailed molecular mechanisms of digestion and utilization of SPs during development are largely unknown. Here, we report the crystal structure ...

  19. Storage protein-2 as a dependable biochemical index for screening germplasm stocks of the silkworm Bombyx mori (L.)

    OpenAIRE

    Jingade H. Anuradha; Perumal Somasundaram; Vishnupriya, S.; A. Manjula

    2012-01-01

    Storage protein (SP-2) variation was investigated among selected silkworm germplasm stocks representing two major potential sericulture areas of India. The expression levels of storage protein varied among them, as seen in Sodium Dodecylsulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE), which correlated with their geographical origin. The storage protein variation is an inter origin variability and this differential expression of the protein is helpful to tag the robustness of the breed/r...

  20. In vivo analysis of fibroin heavy chain signal peptide of silkworm Bombyx mori using recombinant baculovirus as vector

    International Nuclear Information System (INIS)

    In order to investigate the functional signal peptide of silkworm fibroin heavy chain (FibH) and the effect of N- and C-terminal parts of FibH on the secretion of FibH in vivo, N- and C-terminal segments of fibh gene were fused with enhanced green fluorescent protein (EGFP) gene. The fused gene was then introduced into silkworm larvae and expressed in silk gland using recombinant AcMNPV (Autographa californica multiple nuclear polyhedrosis virus) as vector. The fluorescence of EGFP was observed with fluorescence microscope. FibH-EGFP fusion proteins extracted from silk gland were analyzed by Western blot. Results showed that the two alpha helices within N-terminal 163 amino acid residues and the C-terminal 61 amino acid residues were not necessary for cleavage of signal peptide and secretion of the fusion protein into silk gland. Then the C-terminal 61 amino acid residues were substituted with a His-tag in the fusion protein to facilitate the purification. N-terminal sequencing of the purified protein showed that the signal cleavage site is between position 21 and 22 amino acid residues

  1. Functional study on the mutations in the silkworm (Bombyx mori) acetylcholinesterase type 1 gene (ace1) and its recombinant proteins.

    Science.gov (United States)

    Wang, Ju-mei; Wang, Bin-bin; Xie, Yi; Sun, Shan-shan; Gu, Zhi-ya; Ma, Lie; Li, Fan-chi; Zhao, Yi-fan; Yang, Bin; Shen, Wei-de; Li, Bing

    2014-01-01

    The acetylcholinesterase of Lepidoptera insects is encoded by two genes, ace1 and ace2. The expression of the ace1 gene is significantly higher than that of the ace2 gene, and mutations in ace1 are one of the major reasons for pesticide resistance in insects. In order to investigate the effects of the mutations in ace1's characteristic sites on pesticide resistance, we generated mutations for three amino acids using site-directed mutagenesis, which were Ala(GCG)303Ser(TCG), Gly(GGA)329Ala(GCA) and Leu (TCT)554Ser(TTC). The Baculovirus expression system was used for the eukaryotic expression of the wild type ace1 (wace1) and the mutant ace1 (mace1). SDS-PAGE and Western blotting were used to detect the targeting proteins with expected sizeof about 76 kDa. The expression products were purified for the determination of AChE activity and the inhibitory effects of physostigmine and phoxim. We observed no significant differences in the overall activity of the wild type and mutant AChEs. However, with 10 min of physostigmine (10 μM) inhibition, the remaining activity of the wild type AChE was significantly lower than that of the mutant AChE. Ten min inhibition with 33.4 μM phoxim also resulted in significantly lower remaining activity of the wild type AChE than that of the mutant AChE. These results indicated that mutations for the three amino acids reduced the sensitivity of AChE to physostigmine and phoxim, which laid the foundation for future in vivo studies on AChE's roles in pesticide resistance. PMID:24323194

  2. Formation and reparation of the AP-sites into DNA from the gamma-irradiated embryo of bombyx mori

    International Nuclear Information System (INIS)

    Full text: It is well known that radiosensitivity of an organism is in dependence on the DNA reparation systems functioning into cells. Sharp difference in the radioresistance of silkworm embryo at different stage of growth showed by us earlier (Agaev F.A. et al, 1991) can provide to suggest that DNA reparation system into cells of 3-daily embryo (more radiosensitive stage) and 7-daily embryo (more radioresistance stage) may be functioning with various efficiency. It was shown that quantity of the AP-sites (i.e. apurine and apirimidine sites) registered into DNA of 3-daily embryo is 1,2 - 1,4 time more, that into DNA of 7-daily embryo g-irradiated at the same dozes. The increasing of the difference between the registered AP-sites into DNA of 3- and 7-daily embryo has been observed also at the increasing of the radiation doze. At the postradiation incubation of the 3- and 7- daily embryo the lowering of AP-sites quantity into DNA was observed. This fact allowed to testify that the reparation system of damages, such as DNA-apurinization and apirimidinization are functioned into these embryo cells. At the same time the rate of the AP-sites reparation into embryo cells is varied. For example, the remanent quantity of AP-sites into DNA of 7-daily embryo after 45 min of postradiation period consists of 30% those registered immediately after embryo irradiation. The remanent quantity of AP-sites into DNA of 3-daily embryo is lowered on 50%. The difference in the rate of cells reparation is keeping at the constant level irrespective of g-irradiation doze. The binding reaction between the /14 centigrade/-methoxyamine and AP-sites in DNA in vitro has been showed that the reparation time of the 50% AP-sites for 3-daily embryo is 45 min and for 7-daily embryo is 30 min in respective to registered value at once after 100 Gy irradiation doze. In spite of essential difference in the both AP-sites formation into DNA of 3- and 7-daily embryo at once after irradiation and the kinetics its reparation during postradiation period, the results allowed to concluded that such differences cant reflect the 10-fold difference in these embryo sensitivity to g-radiation, determined by the embryo death

  3. Inhibition of BmNPV replication in Bombyx mori cell by dsRNA triggered RNA interference

    Institute of Scientific and Technical Information of China (English)

    XU Ying; ZHU Chenggang; JIN Yongfeng; ZHANG Yaozhou

    2004-01-01

    RNA interference (RNAi) causes degradation of targeted endogenous RNA in many diverse organisms, To investigate the effect of dsRNA on silkworm cells, we transfected three kinds of synthetic dsRNAs of 435 bp(Ap1), 300bp(Ape) and 399 bp(Au) in length against the various regions of BmNPV's DNA polymerase gene and DNA helicase gene,which are indispensable for viral replication in silkworm cells by TransMessengerTM transfection Reagent. Results indicated that in the experiment where silkworm cells were infected with wild-strain BmNPV of the three dsRNAs, Ap2 and AH can effectively suppress the replication of virus, but Ap1 had no effect on the inhibition of viral replication. Ap2 and Au can reduce the infective titer of BmNPV with a peak change of approximately 3-4 logs on day 4 post-infection.The results of reverse transcript polylnerase chain reaction (RT-PCR) and DNA dot blotting also indicated that the expression level of the two target genes and the quantity of viral DNA both distinctly decreased under the influence of Ap2 or Au. Furthermore, using fluorescence microscopy we analyzed the distribution patterns of dsRNA. Our studies revealed that a majority of dsRNA was localized in the nuclear periphery discontinuously after 24 h of transfection.

  4. 2A self-cleaving peptide-based multi-gene expression system in the silkworm Bombyx mori

    Science.gov (United States)

    Wang, Yuancheng; Wang, Feng; Wang, Riyuan; Zhao, Ping; Xia, Qingyou

    2015-01-01

    Fundamental and applied studies of silkworms have entered the functional genomics era. Here, we report a multi-gene expression system (MGES) based on 2A self-cleaving peptide (2A), which regulates the simultaneous expression and cleavage of multiple gene targets in the silk gland of transgenic silkworms. First, a glycine-serine-glycine spacer (GSG) was found to significantly improve the cleavage efficiency of 2A. Then, the cleavage efficiency of six types of 2As with GSG was analyzed. The shortest porcine teschovirus-1 2A (P2A-GSG) exhibited the highest cleavage efficiency in all insect cell lines that we tested. Next, P2A-GSG successfully cleaved the artificial human serum albumin (66 kDa) linked with human acidic fibroblast growth factor (20.2 kDa) fusion genes and vitellogenin receptor fragment (196 kD) of silkworm linked with EGFP fusion genes, importantly, vitellogenin receptor protein was secreted to the outside of cells. Furthermore, P2A-GSG successfully mediated the simultaneous expression and cleavage of a DsRed and EGFP fusion gene in silk glands and caused secretion into the cocoon of transgenic silkworms using our sericin1 expression system. We predicted that the MGES would be an efficient tool for gene function research and innovative research on various functional silk materials in medicine, cosmetics, and other biomedical areas. PMID:26537835

  5. 2A self-cleaving peptide-based multi-gene expression system in the silkworm Bombyx mori

    OpenAIRE

    Yuancheng Wang; Feng Wang; Riyuan Wang; Ping Zhao; Qingyou Xia

    2015-01-01

    Fundamental and applied studies of silkworms have entered the functional genomics era. Here, we report a multi-gene expression system (MGES) based on 2A self-cleaving peptide (2A), which regulates the simultaneous expression and cleavage of multiple gene targets in the silk gland of transgenic silkworms. First, a glycine-serine-glycine spacer (GSG) was found to significantly improve the cleavage efficiency of 2A. Then, the cleavage efficiency of six types of 2As with GSG was analyzed. The sho...

  6. Bombyx mori silk protein films microprocessing with a nanosecond ultraviolet laser and a femtosecond laser workstation: theory and experiments

    Science.gov (United States)

    Lazare, S.; Sionkowska, A.; Zaborowicz, M.; Planecka, A.; Lopez, J.; Dijoux, M.; Louména, C.; Hernandez, M.-C.

    2012-01-01

    Laser microprocessing of several biopolymers from renewable resources is studied. Three proteinic materials were either extracted from the extracellular matrix like Silk Fibroin/Sericin and collagen, or coming from a commercial source like gelatin. All can find future applications in biomedical experimentation, in particular for cell scaffolding. Films of ˜hundred of microns thick were made by aqueous solution drying and laser irradiation. Attention is paid to the properties making them processable with two laser sources: the ultraviolet and nanosecond (ns) KrF (248 nm) excimer and the infrared and femtosecond (fs) Yb:KGW laser. The UV radiation is absorbed in a one-photon resonant process to yield ablation and the surface foaming characteristics of a laser-induced pressure wave. To the contrary, resonant absorption of the IR photons of the fs laser is not possible and does not take place. However, the high field of the intense I>˜1012 W/cm2 femtosecond laser pulse ionizes the film by the multiphoton absorption followed by the electron impact mechanism, yielding a dense plasma capable to further absorb the incident radiation of the end of the pulse. The theoretical model of this absorption is described in detail, and used to discuss the presented experimental effects (cutting, ablation and foaming) of the fs laser. The ultraviolet laser was used to perform simultaneous multiple spots experiments in which energetic foaming yields melt ejection and filament spinning. Airborne nanosize filaments "horizontally suspended by both ends" (0.25 μm diameter and 10 μm length) of silk biopolymer were observed upon irradiation with large fluences.

  7. Analysis of the sericin1 promoter and assisted detection of exogenous gene expression efficiency in the silkworm Bombyx mori L.

    OpenAIRE

    Ye, Lupeng; Qian, Qiujie; Zhang, Yuyu; You, Zhengying; Che, Jiaqian; Song, Jia; Zhong, Boxiong

    2015-01-01

    In genetics, the promoter is one of the most important regulatory elements controlling the spatiotemporal expression of a target gene. However, most studies have focused on core or proximal promoter regions, and information on regions that are more distant from the 5′-flanking region of the proximal promoter is often lacking. Here, approximately 4-kb of the sericin1 (Ser1) promoter was predicted to contain many potential transcriptional factor binding sites (TFBSs). Transgenic experiments hav...

  8. Development of a Foot-and-Mouth Disease Virus Serotype A Empty Capsid Subunit Vaccine Using Silkworm (Bombyx mori) Pupae

    OpenAIRE

    Li, Zhiyong; Yi, Yongzhu; Yin, Xiangping; Zhang, Yun; Liu, Ming; Liu, Hang; Li, Xuerui; Li, Yinü; Zhang, Zhifang; Liu, Jixing

    2012-01-01

    Foot-and-mouth disease (FMD) is a highly contagious disease of cloven-hoofed animals that inflicts severe economic losses in the livestock industry. In 2009, FMDV serotype A caused outbreaks of FMD in cattle in China. Although an inactivated virus vaccine has proven effective to control FMD, its use may lead to new disease outbreaks due to a possible incomplete inactivation of the virus during the manufacturing process. Here, we expressed the P1-2A and the 3C coding regions of a serotype A FM...

  9. Preparation and Characterization of a Novel Hybrid Hydrogel Composed of Bombyx mori Fibroin and Poly(N-isopropylacrylamide)

    International Nuclear Information System (INIS)

    A novel hybrid hydrogel was prepared and investigated based on silkworm silk fibroin and poly(N-isopropylacrylamide) (PNIPAAm). PNIPAAm was introduced to silk fibroin, the resultant composite hydrogel was examined, and freeze-dried SF/PNIPAAm scaffold was analyzed using LB-550 dynamic light scattering particle-size analyzer, circular dichroism (CD), and scanning electron microscopy (SEM). Our results suggested that the hybrid hydrogels owned the porous sponge-like structures, and the gelation time of SF/PNIPAAm hybrids decreased with an increase in temperature and concentration of each polymer. Results of rheological analysis suggested that the rheological property of resultant SF/PNIPAAm gel depended on the concentration combinations as well as the aging time, which elapsed after mixing the two polymers. Results of CD spectra demonstrated that ph showed little influence on the secondary structure of silk fibroin, and significant changes of G', Gand G* as surrounding increase temperature above the lower critical solution temperature (LCST)

  10. Effect of exogenous hormones on transcription levels of pyridoxal 5'-phosphate biosynthetic enzymes in the silkworm (Bombyx mori).

    Science.gov (United States)

    Huang, ShuoHao; Yang, HuanHuan; Yao, LiLi; Zhang, JianYun; Huang, LongQuan

    2016-01-01

    Vitamin B6 includes 6 pyridine derivatives, among which pyridoxal 5'-phosphate is a coenzyme for over 140 enzymes. Animals acquire their vitamin B6 from food. Through a salvage pathway, pyridoxal 5'-phosphate is synthesized from pyridoxal, pyridoxine or pyridoxamine, in a series of reactions catalyzed by pyridoxal kinase and pyridoxine 5'-phosphate oxidase. The regulation of pyridoxal 5'-phospahte biosynthesis and pyridoxal 5'-phospahte homeostasis are at the center of study for vitamin B6 nutrition. How pyridoxal 5'-phosphate biosynthesis is regulated by hormones has not been reported so far. Our previous studies have shown that pyridoxal 5'-phosphate level in silkworm larva displays cyclic developmental changes. In the current study, effects of exogenous juvenile hormone and molting hormone on the transcription level of genes coding for the enzymes involved in the biosynthesis of pyridoxal 5'-phospahte were examined. Results show that pyridoxal kinase and pyridoxine 5'-phosphate oxidase are regulated at the transcription level by development and are responsive to hormones. Molting hormone stimulates the expression of genes coding for pyridoxal kinase and pyridoxine 5'-phosphate oxidase, and juvenile hormone appears to work against molting hormone. Whether pyridoxal 5'-phosphate biosynthesis is regulated by hormones in general is an important issue for further studies. PMID:26780217

  11. Deletion of a gene encoding an amino acid transporter in the midgut membrane causes resistance to a Bombyx parvo-like virus.

    Science.gov (United States)

    Ito, Katsuhiko; Kidokoro, Kurako; Sezutsu, Hideki; Nohata, Junko; Yamamoto, Kimiko; Kobayashi, Isao; Uchino, Keiro; Kalyebi, Andrew; Eguchi, Ryokitsu; Hara, Wajiro; Tamura, Toshiki; Katsuma, Susumu; Shimada, Toru; Mita, Kazuei; Kadono-Okuda, Keiko

    2008-05-27

    Bombyx mori densovirus type 2 (BmDNV-2), a parvo-like virus, replicates only in midgut columnar cells and causes fatal disease. The resistance expressed in some silkworm strains against the virus is determined by a single gene, nsd-2, which is characterized as nonsusceptibility irrespective of the viral dose. However, the responsible gene has been unknown. We isolated the nsd-2 gene by positional cloning. The virus resistance is caused by a 6-kb deletion in the ORF of a gene encoding a 12-pass transmembrane protein, a member of an amino acid transporter family, and expressed only in midgut. Germ-line transformation with a wild-type transgene expressed in the midgut restores susceptibility, showing that the defective membrane protein is responsible for resistance. Cumulatively, our data show that the membrane protein is a functional receptor for BmDNV-2. This is a previously undescribed report of positional cloning of a mutant gene in Bombyx and isolation of an absolute virus resistance gene in insects. PMID:18495929

  12. PANDANGAN MORI OGAI TERHADAP EUTHANASIA (ANRAKUSHI) DALAM TAKASE BUNE

    OpenAIRE

    Linda Unsriana

    2010-01-01

    Mori Ogai is a welknown Japanese author in the modern Japanese literature, that is literatures that reflect a modern society living. This modern society tries to disclose social status and raise freedom and righyt equality as the basic of modern life. One of his novels, Takase Bune clarifies Mori Ogai views on Euthanasia (Anrakushi). Article presents the views of a prominent writer as well as medical practitioner on the Japanese Army on Euthanasia practice.

  13. PANDANGAN MORI OGAI TERHADAP EUTHANASIA (ANRAKUSHI DALAM TAKASE BUNE

    Directory of Open Access Journals (Sweden)

    Linda Unsriana

    2010-03-01

    Full Text Available Mori Ogai is a welknown Japanese author in the modern Japanese literature, that is literatures that reflect a modern society living. This modern society tries to disclose social status and raise freedom and righyt equality as the basic of modern life. One of his novels, Takase Bune clarifies Mori Ogai views on Euthanasia (Anrakushi. Article presents the views of a prominent writer as well as medical practitioner on the Japanese Army on Euthanasia practice.

  14. On the general elephant conjecture for Mori conic bundles

    CERN Document Server

    Prokhorov, Yu G

    1996-01-01

    Let $f:X\\to S$ be an extremal contraction from a threefolds with terminal singularities onto a surface (so called Mori conic bundle). We study some particular cases of such contractions: quotients of usual conic bundles and index two contractions. Assuming Reid's general elephants conjecture we also obtain a rough classification. We present many examples.

  15. MicroRNA of the fifth-instar posterior silk gland of silkworm identified by Solexa sequencing

    Directory of Open Access Journals (Sweden)

    Jisheng Li

    2014-12-01

    Full Text Available No special studies have been focused on the microRNA (miRNA in the fifth-instar posterior silk gland of Bombyx mori. Here, using next-generation sequencing, we acquired 93.2 million processed reads from 10 small RNA libraries. In this paper, we tried to thoroughly describe how our dataset generated from deep sequencing which was recently published in BMC genomics. Results showed that our findings are largely enriched silkworm miRNA depository and may benefit us to reveal the miRNA functions in the process of silk production.

  16. Subcutaneous abscess due to the basidiomycete Phellinus mori in a patient with chronic granulomatous disease.

    Science.gov (United States)

    Shigemura, T; Nakazawa, Y; Amano, Y; Sudo, A; Watanabe, M; Kobayashi, M; Kobayashi, N; Koike, K; Agematsu, K; Nishimura, K

    2015-06-01

    Chronic granulomatous disease (CGD), a primary immunodeficiency caused by impaired phagocyte killing of intracellular pathogens, is characterized by recurrent, life-threatening, bacterial and fungal infections. As a result of improvements in microbiologic culture and identification techniques, a number of unique filamentous fungi have been reported as significant pathogens in patients with CGD. We report a case of subcutaneous basidiomycete Phellinus mori infection in a patient with CGD. To the best of our knowledge, this is the first reported case of human infection by this fungus. The causative fungus was identified on the basis of its morphological characteristics and nucleotide sequence on the internal transcribed spacer region of the ribosomal RNA gene. This is the fifth case report of filamentous basidiomycetes infecting a patient with CGD; all of these cases have been caused by Phellinus species. We highlight the importance of recognizing filamentous basidiomycetes Phellinus species as possible agents of non-Aspergillus fungal infections in patients with CGD. PMID:25600930

  17. The toxicity of NaF on BmN cells and a comparative proteomics approach to identify protein expression changes in cells under NaF-stress

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Liang; Chen, Huiqing [Institute of Life Sciences, Jiangsu University, Zhenjiang, Jiangsu 212013 (China); Yao, Chun [Department of Stomatology, Zhenjiang First People’s Hospital, Zhenjiang, Jiangsu 212013 (China); Chang, Cheng; Xia, Hengchuan; Zhang, Chunxia; Zhou, Yang; Yao, Qin [Institute of Life Sciences, Jiangsu University, Zhenjiang, Jiangsu 212013 (China); Chen, Keping, E-mail: kpchen@ujs.edu.cn [Institute of Life Sciences, Jiangsu University, Zhenjiang, Jiangsu 212013 (China)

    2015-04-09

    Highlights: • On the cellular level, we identified IC{sub 50} of NaF on BmN cell by flow cytometry. • High concentration of NaF gives effect on BmN cell morphology. • Five significantly differential proteins were identified by two-dimensional electrophoresis and mass spectrometry. • ALDH2 and WPH were up-regulated, while CRT and SCF were down-regulated, providing new information for metabolic pathway of fluoride. - Abstract: Fluorides negatively affect the development of organisms and are a threat to human health and environmental safety. In this study, Bombyx mori N cell line (BmN) were used to explore effects of NaF on insect cells. We found that 8 h (hrs) culture with high concentration of NaF (≥1 mM) induced significantly morphological changes. Dose-response curves of 72 h continuously cultured BmN treated with NaF showed that the half inhibitory concentration (IC{sub 50}) value was 56.60 μM. Treatment of BmN with 100 and 300 μM of NaF induced apoptosis and necrosis. 2-D electrophoresis of whole cell extracted from BmN showed that treatment with 300 μM NaF up-regulated 32 proteins and down-regulated 11 proteins when compared with controls. We identified 5 different proteins by MALDI-TOF MS, and 4 of them were identified for the first time, including 2 up-regulated proteins (mitochondrial aldehyde dehydrogenase ALDH2 and prohibitin protein WPH) and 2 down-regulated proteins (calreticulin precursor CRT and DNA supercoiling factor SCF). These observations were further confirmed by fluorescence quantitative PCR. Together, our data suggest that these target proteins could be regarded as targets influenced by NaF and also provide clues for studies on the response metabolism pathway under NaF stress.

  18. The toxicity of NaF on BmN cells and a comparative proteomics approach to identify protein expression changes in cells under NaF-stress

    International Nuclear Information System (INIS)

    Highlights: • On the cellular level, we identified IC50 of NaF on BmN cell by flow cytometry. • High concentration of NaF gives effect on BmN cell morphology. • Five significantly differential proteins were identified by two-dimensional electrophoresis and mass spectrometry. • ALDH2 and WPH were up-regulated, while CRT and SCF were down-regulated, providing new information for metabolic pathway of fluoride. - Abstract: Fluorides negatively affect the development of organisms and are a threat to human health and environmental safety. In this study, Bombyx mori N cell line (BmN) were used to explore effects of NaF on insect cells. We found that 8 h (hrs) culture with high concentration of NaF (≥1 mM) induced significantly morphological changes. Dose-response curves of 72 h continuously cultured BmN treated with NaF showed that the half inhibitory concentration (IC50) value was 56.60 μM. Treatment of BmN with 100 and 300 μM of NaF induced apoptosis and necrosis. 2-D electrophoresis of whole cell extracted from BmN showed that treatment with 300 μM NaF up-regulated 32 proteins and down-regulated 11 proteins when compared with controls. We identified 5 different proteins by MALDI-TOF MS, and 4 of them were identified for the first time, including 2 up-regulated proteins (mitochondrial aldehyde dehydrogenase ALDH2 and prohibitin protein WPH) and 2 down-regulated proteins (calreticulin precursor CRT and DNA supercoiling factor SCF). These observations were further confirmed by fluorescence quantitative PCR. Together, our data suggest that these target proteins could be regarded as targets influenced by NaF and also provide clues for studies on the response metabolism pathway under NaF stress

  19. Mori Folium and Mori Fructus Mixture Attenuates High-Fat Diet-Induced Cognitive Deficits in Mice

    Directory of Open Access Journals (Sweden)

    Hyo Geun Kim

    2015-01-01

    Full Text Available Obesity has become a global health problem, contributing to various diseases including diabetes, hypertension, cancer, and dementia. Increasing evidence suggests that obesity can also cause neuronal damage, long-term memory loss, and cognitive impairment. The leaves and the fruits of Morus alba L., containing active phytochemicals, have been shown to possess antiobesity and hypolipidemic properties. Thus, in the present study, we assessed their effects on cognitive functioning in mice fed a high-fat diet by performing immunohistochemistry, using antibodies against c-Fos, synaptophysin, and postsynaptic density protein 95 and a behavioral test. C57BL/6 mice fed a high-fat diet for 21 weeks exhibited increased body weight, but mice coadministered an optimized Mori Folium and Mori Fructus extract mixture (2 : 1; MFE for the final 12 weeks exhibited significant body weight loss. Additionally, obese mice exhibited not only reduced neural activity, but also decreased presynaptic and postsynaptic activities, while MFE-treated mice exhibited recovery of these activities. Finally, cognitive deficits induced by the high-fat diet were recovered by cotreatment with MFE in the novel object recognition test. Our findings suggest that the antiobesity effects of MFE resulted in recovery of the cognitive deficits induced by the high-fat diet by regulation of neural and synaptic activities.

  20. On the Breeding of Bivoltine Double Hybrid of Silkworm Bombyx mori (Lepidoptera: Bombycidae) Tolerant to High Temperature and Low Humidity Conditions of the Tropics

    OpenAIRE

    2010-01-01

    The hot climatic conditions of tropics prevailing particularly in summer are contributing to the poor performance of the bivoltine breeds and the most important aspect is that many quantitative characters such as viability and cocoon traits decline sharply when temperature is high. Hence, in a tropical country like India, it is very essential to develop bivoltine breeds/hybrids which can withstand the high temperature stress conditions. This has resulted in the development of CSR18 × CSR19, c...

  1. 家蚕人工授精关键技术的研究%Study on Key Technique of Artificial Insemination for Silkworm, Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    张业顺; 张国政; 韦亚东; 夏定国

    2009-01-01

    [Objective] The aim of this study was to investigate the efficient technique of artificial insemination for silkworm. [Method] Sperms were extracted from bursa copulatrix of female moths mated for 30 min through extruding and centrifugal method, and then the semen was injected into other virgin moths with trypsinase. [Result] A high-effective collection technology of spermatids from silkworm was established successfully, 50 μl semen could be collected by only one person in each hour. The survival rate of spermatids was over 80% in vito after collected from bursa copulatrix, while the obtained semen was quite pure and the average fertilization rate of silkworm was 76.5%. [Conclusion] The establishment of high-effective semen extraction technique of silkworm provides the technical basis for studies on other related techniques for silkworm sperm.

  2. Immune response in cattle inoculated with the recombinant complete polyprotein of foot-and-mouth disease virus from Bombyx mori larvae

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    The intact open reading frame (ORF) of foot-and-mouth disease virus (FMDV) Asia I/XJ strain was amplified by RT-PCR and inserted into the transfer vector pVL1393 to generate plasmid pVL-ORF. Bm-N cells were transfected with pVL-ORF and linearized Bm-BacPAK6 DNA, and the recombinant silkworm baculovirus Bm-ORF containing the full ORF of FMDV was obtained. The results of indirect immunofluorescence assay (IFA) showed that Bm-ORF could be expressed efficiently in Bm-N cell. After inoculating the early 5th instar larvae of silkworm, the polyprotein of FMDV could be detected by sandwich ELISA and empty capsid-like particles could be observed under the electron microscope. Expression products from silkworm were used as the antigen to immunize the cattle. The specific antibody was induced in all vaccinated animals. The immunized cattle were challenged with the virulent FMDV Asia I/XJ strain, two of the four cattle were completely protected and clinical symptoms were alleviated and delayed in the others. The results suggest that this strategy might be used to develop the new subunit FMDV vaccine.

  3. Studies on middle silkgland proteins of cocoon colour sex-limited silkworm (Bombyx mori L.) using two-dimensional polyacrylamide gel electrophoresis

    Indian Academy of Sciences (India)

    Yuan-Xiang Jin; Yu-Yin Chen; Meng-Kui Xu; Yong-Huang Jiang

    2004-03-01

    Qualitative and quantitative differences in proteins expressed in the middle silkglands of male and female silkworm larvae that differ in silk colour were investigated by high resolution two-dimensional polyacrylamide gel electrophoresis (2-D PAGE), followed by computer assisted image analysis. About 1000 protein spots were resolved in both the sexes and most proteins were shown to be distributed in the area from 15 kDa to 70 kDa and pH 4–8. It was found that some proteins displayed higher expression in yellow cocoon, while two proteins were only expressed in female silkworm silkgland tissue through the comparison and analysis by two-D software. These proteins especially existed in female silkworm middle silkgland tissue of yellow cocoon. Furthermore, these proteins might be involved in the expression of cocoon colour phenotype.

  4. Contribution to the ultrastructural study of silk-excretion cells and autoradiographic analysis of intracellular fibroin transport in Bombyx mori L

    International Nuclear Information System (INIS)

    It is much easier to study the mechanisms involved in the synthesis and exportation of extracellular proteins in the biological material chosen is highly differentiated. The silk-excretion gland of the silkworm is ideal in this respect because during the larva period, especially at the end of the 5th and last stage, the cells at the rear (excreting tube) synthesize and export massive quantities of a single protein: fibroin. These phenomena were explored by a cytological study carried out mainly by electron microscopy and autoradiography. The results obtained are given. They relate first of all to the morphological development of the secretion tube cells from the end of the 4th larva stage to the spinning of the cocoon, and contribute new information on the cell changes during the 4th slough and the end of the 5th age. They also concern intracellular fibroin transport which is proved to take place through the Golgi apparatus, and finally the possible role of the microtubules and microfilaments in fibroin transport and secretion. On this last point the results so far constitute only, a preliminary approach which justifie no final conclusions; they merely suggest that the microfilaments of the apical region are involved in the secretion process

  5. Development of Bivoltine Pure Strains of Silkworm, Bombyx mori L. to Rear Eexclusively on Artificial Diet During Young Instar

    OpenAIRE

    K.S. Nair; S.N. Kumar; J.S. Nair

    2011-01-01

    As component bivoltine pure strains of existing productive silkworm hybrids did not accept the artificial diet, the study was taken up to create a pool of such strains which would feed on the artificial diet. Six strains viz., 5HT, GEN4, 8HT, B71, CSR3 and JPN8 were short-listed based on the results of an initial screening which recorded feeding response percentage of more than 20. These six strains were further subjected to continuous inbreeding and directional selection for 12 generations f...

  6. Genetic Analysis of Izoenzymes Polymorphisms in Silkworm (Bombyx mori L.) Strains - doi: 10.4025/actascibiolsci.v35i2.13102

    OpenAIRE

    Maria Aparecida Fernandez; Ludimilla Ronqui; Maria Claudia Colla Ruvolo Takasusuki

    2013-01-01

    This work carried out to evaluates the polymorhism in the silkworm of different lineages using the isoenzymes electrophoresis to detect biochemical markers and to investigate the genetics of populations for those lineages. They were used as samples individual extracts of silk glands of second day old larvas of the fifth instar, originating from seven Japanese lineages and eight pure Chinese lineages maintained by the Cocamar-Cooperativa Agroindustrial de Maringá. The isozymes acid phosphatase...

  7. Peptidoglycan recognition protein-S5 functions as a negative regulator of the antimicrobial peptide pathway in the silkworm, Bombyx mori.

    Science.gov (United States)

    Chen, Kangkang; Zhou, Lin; Chen, Feng; Peng, Yachun; Lu, Zhiqiang

    2016-08-01

    Prophenoloxidase (proPO), immune deficiency (IMD), and Toll are the major signaling pathways leading to melanization and antimicrobial peptide production in insect hemolymph. Peptidoglycan recognition proteins (PGRPs) act as receptors and negative regulators in these pathways, and some PGRPs exhibit antimicrobial activity. Previously, we demonstrated that silkworm PGRP-S5 recognizes peptidoglycans (PGs) and triggers activation of the proPO pathway. It also acts as a bactericide, via its amidase activity (Chen et al., 2014). Here, we generated a C177S site-mutated silkworm PGRP-S5 protein that lacked amidase activity but retained its PG-binding capacity. Functional studies showed that the mutation caused loss of its receptor function for activation of the proPO pathway, suggesting that processing of PG by PGRP-S5 is necessary for formation of the pathway initiation complex. Further, we found that PGRP-S5 negatively regulates antimicrobial peptides generation in an amidase-dependent manner, likely through the IMD pathway. Thus, silkworm PGRP-S5 acts as a sensor, a modulator, and an effector in the silkworm humoral immune system. PMID:27012996

  8. On the Breeding of Bivoltine Breeds of the Silkworm, Bombyx mori L. (Lepidoptera: Bombycidae, Tolerant to High Temperature and High Humidity Conditions of the Tropics

    Directory of Open Access Journals (Sweden)

    Harjeet Singh

    2010-01-01

    Full Text Available The hot climatic conditions of tropics prevailing particularly in summer are contributing to the poor performance of the bivoltine breeds and the most important aspect is that many quantitative characters such as viability and cocoon traits decline sharply when temperature is high. Hence, in a tropical country like India, it is very essential to develop bivoltine breeds/hybrids which can withstand the high temperature stress conditions. This has resulted in the development of CSR18 × CSR19, compatible hybrid for rearing throughout the year by utilizing Japanese thermotolerant hybrids as breeding resource material. Though, the introduction of CSR18 × CSR19 in the field during summer months had considerable impact, the productivity level and returns realized do not match that of other productive CSR hybrids. Therefore, the acceptance level of this hybrid with the farmers was not up to the expected level. This has necessitated the development of a temperature tolerant hybrid with better productivity traits than CSR18 × CSR19. Though, it was a difficult task to break the negative correlation associated with survival and productivity traits, attempts on this line had resulted in the development of CSR46 × CSR47, a temperature tolerant bivoltine hybrid with better productivity traits than CSR18 × CSR19. However, though, these hybrids are tolerant to high temperature environments, they are not tolerant to many of the silkworm diseases. Keeping this in view, an attempt is made to develop silkworm hybrids tolerant to high temperature environments.

  9. Effect of ATG initiation codon context motifs on the efficiency of translation of mRNA derived from exogenous genes in the transgenic silkworm, Bombyx mori

    OpenAIRE

    Tatematsu, Ken-ichiro; Uchino, Keiro; Sezutsu, Hideki; Tamura, Toshiki

    2014-01-01

    The context sequence motif surrounding the ATG initiation codon influences mRNA translation efficiency and affects protein production; however, the optimal sequence differs among species. To determine the optimal sequence for production of recombinant proteins in a transgenic silkworm, we compared 14-nucleotide context motifs around the ATG (ATG-context) in 50 silkworm genes and found the following consensus: (A/T)AN(A/T)ATCAAAatgN. We were also able to define the least-common motif: CCN(C/G)...

  10. Investigation of the Mutant Character Flossy in the Silkworm, Bombyx mori%家蚕绵茧突变性状调查

    Institute of Scientific and Technical Information of China (English)

    刘春; 朱晓楠; 刘茹凤; 任静波; 石虎; 荀立杰; 夏庆友

    2012-01-01

    家蚕绵茧(Flossy)突变系二化性、单基因显性突变,表型为茧形浮松、多分层,是几个较为重要的蚕茧突变之一。本文选取夏芳、大造作为对照,对绵茧的茧层率、含胶率、茧丝显微镜观察、丝胶蛋白SDS-PAGE进行调查比较。实验结果显示,夏芳、大造、绵茧茧层率为23.476%、12.919%、15.551%;含胶率依次为24.980%、33.765%、28.399%;夏芳、大造、绵茧茧层显微镜观察结果未显示明显差异;丝胶蛋白成分分析结果显示,绵茧和对照之间没有明显不同。就目前结果来看,丝胶蛋白含量有可能是致使绵茧表型的原因,但与对照的差异不明显,因而绵茧突变性状形成的真正原因有待进一步研究。本研究为家蚕绵茧基因的克隆及功能研究提供参考信息。%Silkworm is one of the most important model insects. Further more, it is an important genetic material. Currently, there are more than 400 silkworm mutants preserved in Silk-worm Resource Library. Research of the underlying physiological and biochemical mecha- nisms for the generation of silkworm mutant phenotypes can provide effective clews for posi- tional cloning research. Flossy mutant is a digoneutism, single gene dominant mutant, whose phenotype is characterized by floating loose cocoons with much layering, and Flossy is one of the outbalance silkworm cocoon mutants. In this study, Xiafang and Dazao were cho- sen as the control, the cocoon shell rate and the sericin holding-rate of Flossy were observed microscopically and its sericin protein was analyzed with SDS-PAGE. The results showed that the cocoon shell rate of Xiafang, Dazao and Flossy was 23. 476%, 12. 919% and 15. 551%; and their the sericin holding-rate was 24. 980%, 33. 765% and 28. 399% respec- tively. Microscopic examination revealed no outstanding difference in cocoon shell among Xiafang, Dazao and Flossy. The analysis of sericin proteins also showed little difference be tween Flossy and the control. In conclusion, the level of sericins proteins might be the rea- son for the Flossy phenotype, but the difference between the mutant and the control in this respect was non-significant. Therefore, more research is needed to clarify the exact mecha- nism for the generation of the Flossy mutant character.

  11. Antioxidant activities of two sericin proteins extracted from cocoon of silkworm (Bombyx mori) measured by DPPH, chemiluminescence, ORAC and ESR methods

    OpenAIRE

    TAKECHI, TAYORI; WADA, RITSUKO; FUKUDA, TSUBASA; HARADA, KAZUKI; TAKAMURA, HITOSHI

    2014-01-01

    Recent efforts have focused on the use of sericin proteins extracted from cocoons of silkworm as a healthy food source for human consumption. In this study, we focused on the antioxidative properties of sericin proteins. The antioxidative properties were measured in sericin proteins extracted from the shell of the cocoon, designated hereafter as white sericin protein and yellow-green sericin protein, as well as bread without sericin protein and bread to which white sericin powder had been add...

  12. Binding of phylogenetically distant Bacillus thuringiensis cry toxins to a Bombyx mori aminopeptidase N suggests importance of Cry toxin's conserved structure in receptor binding.

    Science.gov (United States)

    Shinkawa, A; Yaoi, K; Kadotani, T; Imamura, M; Koizumi, N; Iwahana, H; Sato, R

    1999-07-01

    We investigated the binding proteins for three Cry toxins, Cry1Aa, Cry1Ac, and the phylogenetically distant Cry9Da, in the midgut cell membrane of the silkworm. In a ligand blot experiment, Cry1Ac and Cry9Da bound to the same 120-kDa aminopeptidase N (APN) as Cry1Aa. A competition experiment with the ligand blot indicated that the three toxins share the same binding site on several proteins. The values of the dissociation constants of the three Cry toxins and 120-kDa APN are as low as the case of other Cry toxins and receptors. These results suggest that distantly related Cry toxins bind to the same site on the same proteins, especially with APN. We propose that the conserved structure in these three toxins includes the receptor-binding site. PMID:10387111

  13. Intercultural Usage of Mori Folium: Comparison Review from a Korean Medical Perspective

    Directory of Open Access Journals (Sweden)

    Byungjin Joh

    2015-01-01

    Full Text Available Objectives. A review on studies related to the use of Mori folium, the leaves of Morus alba, was conducted with the goal of identifying new clinical applications in Korean medicine. Methods. Global literature search was conducted using three electronic databases up to January 2015 with the term Morus alba and its Korean terms. KM literatures including textbooks and standard pharmacopoeia were separately hand-searched and reviewed to provide comparison. Data were extracted according to predetermined criteria, and clinical uses were standardized with ICD-10 categories. Results. 159 potentially relevant studies were identified, and 18 articles including 12 ethnopharmacologic and 6 clinical studies were finally included in this analysis. Ethnopharmacologic studies from 8 countries provided 17 clinical uses. We found that five out of six clinical trials were related to diabetes and suggested a moderate short-term to mild long-term effect. And 43 Korean texts also provided 156 clinical uses in 35 categories including ocular and respiratory disorders. Discussion and Conclusions. Though majority of the clinical uses were also found in Korean medicine literature, treatment of infertility, jaundice, cognitive disorder, and hyperpigmentation was found to be effective and diabetes with Morus alba was recognized to have clinical importance.

  14. Approximate but accurate quantum dynamics from the Mori formalism: I. Nonequilibrium dynamics

    Science.gov (United States)

    Montoya-Castillo, Andrés; Reichman, David R.

    2016-05-01

    We present a formalism that explicitly unifies the commonly used Nakajima-Zwanzig approach for reduced density matrix dynamics with the more versatile Mori theory in the context of nonequilibrium dynamics. Employing a Dyson-type expansion to circumvent the difficulty of projected dynamics, we obtain a self-consistent equation for the memory kernel which requires only knowledge of normally evolved auxiliary kernels. To illustrate the properties of the current approach, we focus on the spin-boson model and limit our attention to the use of a simple and inexpensive quasi-classical dynamics, given by the Ehrenfest method, for the calculation of the auxiliary kernels. For the first time, we provide a detailed analysis of the dependence of the properties of the memory kernels obtained via different projection operators, namely, the thermal (Redfield-type) and population based (NIBA-type) projection operators. We further elucidate the conditions that lead to short-lived memory kernels and the regions of parameter space to which this program is best suited. Via a thorough analysis of the different closures available for the auxiliary kernels and the convergence properties of the self-consistently extracted memory kernel, we identify the mechanisms whereby the current approach leads to a significant improvement over the direct usage of standard semi- and quasi-classical dynamics.

  15. Approximate but accurate quantum dynamics from the Mori formalism: I. Nonequilibrium dynamics.

    Science.gov (United States)

    Montoya-Castillo, Andrés; Reichman, David R

    2016-05-14

    We present a formalism that explicitly unifies the commonly used Nakajima-Zwanzig approach for reduced density matrix dynamics with the more versatile Mori theory in the context of nonequilibrium dynamics. Employing a Dyson-type expansion to circumvent the difficulty of projected dynamics, we obtain a self-consistent equation for the memory kernel which requires only knowledge of normally evolved auxiliary kernels. To illustrate the properties of the current approach, we focus on the spin-boson model and limit our attention to the use of a simple and inexpensive quasi-classical dynamics, given by the Ehrenfest method, for the calculation of the auxiliary kernels. For the first time, we provide a detailed analysis of the dependence of the properties of the memory kernels obtained via different projection operators, namely, the thermal (Redfield-type) and population based (NIBA-type) projection operators. We further elucidate the conditions that lead to short-lived memory kernels and the regions of parameter space to which this program is best suited. Via a thorough analysis of the different closures available for the auxiliary kernels and the convergence properties of the self-consistently extracted memory kernel, we identify the mechanisms whereby the current approach leads to a significant improvement over the direct usage of standard semi- and quasi-classical dynamics. PMID:27179468

  16. EST Table: NM_001111334 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available NM_001111334 Br-c 10/09/29 81 %/420 aa ref|NP_001104804.1| broad-complex isoform Z1... [Bombyx mori] dbj|BAD23978.1| broad-complex Z1-isoform [Bombyx mori] dbj|BAD23983.1| broad-complex Z1-isofo...rm [Bombyx mori] dbj|BAD24045.1| Broad-Complex isoform Z1 [Bombyx mori] dbj|BAD24046.1| Broad-Complex isofor...m Z1 [Bombyx mori] dbj|BAD46732.1| broad-complex A-Z1 isoform [Bombyx mori] dbj|BAD46739.1| broad...-complex B-Z1 isoform [Bombyx mori] dbj|BAF43564.1| Broad-Complex isoform Z1 [Bombyx mori] 1

  17. Desenvolvimento biológico das raças puras e dos híbridos do bicho-da-seda (Bombyx mori L., após o 5º ínstar, em duas estações do ano Biological development after 5th instar among pure breed and hybrids of silkworm (Bombyx mori L.

    Directory of Open Access Journals (Sweden)

    Roque Takahashi

    2002-04-01

    Full Text Available O objetivo deste experimento foi o de verificar a duração da subida no bosque e formação do casulo; duração da formação do casulo até a emergência da mariposa e longevidade do adulto, assim como o peso médio do casulo verde e teor líquido de seda, entre duas raças puras - AJ1 (Japonesa e BC1 (Chinesa e dois híbridos, originários do cruzamento recíproco entre duas raças puras - BC1 X AJ1 (H1 e AJ1 X BC1 (H2, em duas épocas distintas: primavera (E1 e outono (E2. Os híbridos e BC1 foram superiores à raça pura AJ1 ao confeccionarem os casulos, levando em média 3,4 dias e 4,2 dias, respectivamente (E1. As lagartas criadas na E2, mostraram-se superiores em 77,88% em seus tempos de subida no bosque e confecção do casulo, em relação a E1. Quanto ao parâmetro emergência do adulto, sobressaiu-se o H2 (média de 8,9 dias, na E1, em relação às raças puras. As fêmeas dos híbridos, sobreviveram uma média de 3,8 dias a mais que as raças puras, para a E1. Na E2, o H1 destacou-se em relação às raças puras, cuja longevidade foi superior em 9,6 dias. Para os machos, na E2, a longevidade do H2 (5,0 dias foi superior à raça pura AJ1 (4,0 dias. Os híbridos e a AJ1 apresentaram casulos mais pesados em 15,2% em relação a BC1 (E2 e teor líquido de seda não foi significativo em ambas as épocas.This research was carried out to verify the duration of the rising up of caterpillar and the cocoon formation; duration of the cocoon formation to the emergency of the moth and the life span of the adult, as well as the medium weight of the green cocoon and liquid rate of silk, among two pure breeds - AJ1 (Japanese and BC1 (Chinese and two hybrids, from reciprocal crossing among two pure breeds - BC1 X AJ1 (H1 and AJ1 X BC1 (H2, in two different seasons: spring (S1 and autumn (S2. The hybrids and BC1 breed were superior to the pure race AJ1 making the cocoons, taking on average 3.4 days and 4.2 days, respectively (S1. The caterpillars grew in S2 were superior in 77.88% in their times of rising up and making the cocoon, in relation to S1. The hybrid H2 was better than the pure breeds in emergency of adult with average of 8.9 days, in S1. The females of the hybrid, survived an average of 3.8 more days than the pure breeds, in S1. In S2, H1 stood out in relation to the pure races, whose life span was superior in 9.6 days. For the males, in S2, the life span of H2 was 5.0 days. It was superior to the pure race AJ1 - 4.0 days. The hybrids and AJ1 presented heavier cocoons in 15.2% in relation to BC1 (S2 and liquid rate of silk was not significant in both seasons.

  18. Gene : CBRC-GGOR-01-1406 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available QATQPGKPAKPPSQTASQASQASQASQPPKPAKPPSQPASQARQATQPASQPASQASQATKPGKPAKPPSQPDSQPSQASQASKRSQRAKPAKPASQSPKPAKPASHPSQPSPPAT ... ...ive cuticle protein [Bombyx mori] 3e-82 34% MTCFWGEGQEKSDQRXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXFQPSQPSQPRKPASQASQASQASQQAKPA...QPPSQPSQPSHTSSQASQARRESLPAKPAKPAKPASKPSHPSQPANQASQASQPAKPAKPASQPIQPRQPSKPSQPSQPSQPASQASQATQPGKPAKPPSQTASQASQPPKPA...KPPSQPASQARQATQPAKPAQPPSQPSQPSHPSSQASQASRESLPAKPAKPTSKPSQPSHPARQASQATQPDSQPSQPSQPSQPA...TQASQATKPASQPSQTSHPASQPAKPASQPSQPSHPSSQASPATQPAKPAKPPFQPSQPSHPASQASQATLPAKPAKPAKPAREASQPASQPVKPASQASQASQAS

  19. Gene : CBRC-GGOR-01-0478 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available ive cuticle protein [Bombyx mori] 2e-77 32% MQRPAKPAQAFPTLGPSQPASQTSQASDPPKPAKPASHLSQPSQTASQASQLAIQPSQPSQAASHASQASRPAKPAKPAKPAKPA...QASQANQQAKPAKPPSQASQPSHPARQPAKPAKPVKPASHPSQPSHPASQPAKPDKPPSQPASQPAKPAKPPSQASQPSHPASSQTANPAKPAKPAKEASEPSQPSQQASH...PSQPNQPVTQVSQARQPPKPAKPASHPSQPSQPASKASQASEASQPAKPAKPASQASQASRASQRSQLAKPAQPAQPASQPSQASQPASQASPASQPASKASQAS...QATQPAKPAQSPSQPSQPASQASQASQARQASQATQPAKPANPAKPAKPAKEASEPSQPSQQASH...PSQPNQPVTQVSLARQPPKPAKPASHSSQPNHPASKASQASEASQPAKPAKPASQASQASRASQRSQLAKPAKPAQPAGQPSHKSQPPSQAKPAQPASQQSQPSQPSQPTSQPVKPASQQASQ ...

  20. Gene : CBRC-GGOR-01-1485 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available GQPRQPSHLAGQARQATRPASQPSQRSKPSQPAGRASQPAKTAKPPSRQSQPSHTASQARLATLPPKPAKLPSQSIQPSHPTSQPATPASHPSQPATQASQAAQSAS...QPSQPRDPASQANQATKPGKPASQPSQPRQPAKSASQVXXXXKKPASKPREPSQPTWPASEASQASKPAKPAKTASHPSKASRPA...LAGQARQATQPASQPSQRSKPSQPAGRASQPAKTAKPPGGPSQASHPASQPASQPSRPSQPAGQASQPAKTAKPPSRQSQPSHTASQARLATLPPKPAKPPSQSSQPSHPVIKAS...KPAGQPSQQSHSANQASQSTRPAQPGKTAKPPGQPCQPSQSARQASQASQASQARQPSQPSQPSQAAKPASQPSQTSQPASQASQAIQPDKPAGQPRQPSH...ive cuticle protein [Bombyx mori] 4e-64 31% MPAKPVSQAGQPSQPSQPTSQASQPANQPNQPSQPASLATLASQPAKPTKPGTQASQASQASPTSQTSQPARQASQAIQPDKPA

  1. Hierarchical Modeling of Mastic Asphalt in Layered Road Structures Based on the Mori-Tanaka Method

    Directory of Open Access Journals (Sweden)

    Richard Valenta

    2012-01-01

    Full Text Available We present an application of the Mori-Tanaka micromechanical model for a description of the highly nonlinear behavior of asphalt mixtures. This method is expected to replace an expensive finite element-based fully-coupled multi-scale analysis while still providing useful information about local fields on the meso-scale that are not predictable by strictly macroscopic simulations. Drawing on our recent results from extensive experimental and also numerical investigations this paper concentrates on principal limitations of the Mori-Tanaka method, typical of all two-point averaging schemes, when appliedto material systems prone to evolving highly localized deformation patterns such as a network of shear bands. The inability of the Mori-Tanaka method to properly capture the correct stress transfer between phases with increasing compliance of the matrix phase is remedied here by introducing a damage like parameter into the local constitutive equation of reinforcements (stones to control an amount of stress taken by this phase. A deficiency of the Mori-Tanaka method in the prediction of creep response is also mentioned particularly in the light of large scale simulations. A comparison with the application of macroscopic homogenized constitutive model for an asphalt mixture is also presented.

  2. Anti-inflammatory action of ethanolic extract of Ramulus mori on the BLT2-linked cascade.

    Science.gov (United States)

    Park, Geun-Soo; Kim, Jeong-Keun; Kim, Jae-Hong

    2016-04-01

    Mulberry tree twigs (Ramulus mori) contain large amounts of oxyresveratrols and have traditionally been used as herbal medicines because of their anti-inflammatory properties. However, the signaling mechanism by which R. mori exerts its anti-inflammatory action remains to be elucidated. In this study, we observed that R. mori ethanol extracts (RME) exerted an inhibitory effect on the lipopolysaccharide (LPS)-induced production of the pro-inflammatory cytokine interleukin-6 (IL-6) in Raw264.7 macrophage cells. Additionally, RME inhibited IL-6 production by blocking the leukotriene B4 receptor- 2 (BLT2)-dependent-NADPH oxidase 1 (NOX1)-reactive oxygen species (ROS) cascade, leading to anti-inflammatory activity. Finally, RME suppressed the production of the BLT2 ligands LTB4 and 12(S)-HETE by inhibiting the p38 kinase- cytosolic phospholipase A2-5-/12-lipoxygenase cascade in LPS-stimulated Raw264.7 cells. Overall, our results suggest that RME inhibits the 'BLT2 ligand-BLT2'-linked autocrine inflammatory axis, and that this BLT2-linked cascade is one of the targets of the anti-inflammatory action of R. mori. [BMB Reports 2016; 49(4): 232-237]. PMID:26879317

  3. Sensitivity of bombyx embryo during embryonic development to low dose of heavy ion

    International Nuclear Information System (INIS)

    Sensitivity of Bombyx eggs to a heavy ion beam was estimated from the frequency of somatic mutation which was shown by the white spot on the black integument of the fifth instar larvae, indicating the highest on day 2 during resumption of embryogenesis after the termination of diapause, dependently on the dose of Ne, C and Fe ion beams. Irradiation of Fe beams (0.02 Gy) to the eggs induced the somatic mutation and germ cell mutation. (author)

  4. Applied Mori theory of the moduli space of stable pointed rational curves

    OpenAIRE

    Larsen, Paul L.

    2011-01-01

    Diese Dissertation befasst sich mit Fragen über den Modulraum M_{0,n} der stabilen punktierten rationalen Kurven, die durch das Mori-Programm motiviert sind. Insbesondere studieren wir den nef-Kegel (Chapter 2), den Cox-Ring (Chapter 3), und den Kegel der beweglichen Kurven (Chapter 4). In Kapitel 2 beweisen wir Fultons Vermutung für M_{0,n}, n

  5. Two Intracellular Symbiotic Bacteria from the Mulberry Psyllid Anomoneura mori (Insecta, Homoptera)

    OpenAIRE

    Fukatsu, Takema; Nikoh, Naruo

    1998-01-01

    We characterized the intracellular symbiotic bacteria of the mulberry psyllid Anomoneura mori by performing a molecular phylogenetic analysis combined with in situ hybridization. In its abdomen, the psyllid has a large, yellow, bilobed mycetome (or bacteriome) which consists of many round uninucleated mycetocytes (or bacteriocytes) enclosing syncytial tissue. The mycetocytes and syncytium harbor specific intracellular bacteria, the X-symbionts and Y-symbionts, respectively. Almost the entire ...

  6. Hierarchical Modeling of Mastic Asphalt in Layered Road Structures Based on the Mori-Tanaka Method

    OpenAIRE

    Richard Valenta; Michal Šejnoha

    2012-01-01

    We present an application of the Mori-Tanaka micromechanical model for a description of the highly nonlinear behavior of asphalt mixtures. This method is expected to replace an expensive finite element-based fully-coupled multi-scale analysis while still providing useful information about local fields on the meso-scale that are not predictable by strictly macroscopic simulations. Drawing on our recent results from extensive experimental and also numerical investigations this paper concentrate...

  7. In Vivo Bioassay of Recombinant Human Growth Hormone Synthesized in B. mori Pupae

    Directory of Open Access Journals (Sweden)

    Hanglian Lan

    2010-01-01

    Full Text Available The human growth hormone (hGH has been expressed in prokaryotic expression system with low bioactivity previously. Then the effective B. mori baculovirus system was employed to express hGH identical to mature hGH successfully in larvae, but the expression level was still limited. In this work, the hGH was expressed in B. mori pupae by baculovirus system. Quantification of recombinant hGH protein (BmrhGH showed that the expression of BmrhGH reached the level of approximately 890 μg/mL pupae supernatant solution, which was five times more than the level using larvae. Furthermore, Animals were gavaged with BmrhGH at the dose of 4.5 mg/rat.day, and the body weight gain (BWG of treated group had a significant difference (P<.01 compared with the control group. The other two parameters of liver weight and epiphyseal width were also found to be different between the two groups (P<.05. The results suggested that BmrhGH might be used as a protein drug by oral administration.

  8. EST Table: CK507581 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS917189 swd ...

  9. EST Table: CK505540 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h CK504855 swc ...

  10. EST Table: CK505630 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available byx mori] dbj|BAE53373.1| antibacterial peptide [Bombyx mori] dbj|BAF63528.1| gloverin4 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS798027 swc ...

  11. EST Table: FS892293 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ombyx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/09/12 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS866683 ftes ...

  12. EST Table: CK502372 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available byx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h CK504855 swc ...

  13. EST Table: CK496351 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS917189 swb ...

  14. EST Table: CK506703 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS917189 swc ...

  15. EST Table: CK512575 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available byx mori] dbj|BAE53373.1| antibacterial peptide [Bombyx mori] dbj|BAF63528.1| gloverin4 [Bombyx mori] 10/08/31 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS798027 swd ...

  16. EST Table: CK505538 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available byx mori] dbj|BAE53373.1| antibacterial peptide [Bombyx mori] dbj|BAF63528.1| gloverin4 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS798027 swc ...

  17. EST Table: FS898270 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ombyx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/09/12 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS866683 ftes ...

  18. EST Table: DC538251 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ombyx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/09/02 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS917189 MFB- ...

  19. EST Table: CK498951 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS917189 swb ...

  20. EST Table: CK490309 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS917189 swa ...

  1. EST Table: CK506390 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS917189 swc ...

  2. EST Table: CK507467 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h CK504855 swc ...

  3. EST Table: FS798943 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ombyx mori] dbj|BAE53373.1| antibacterial peptide [Bombyx mori] dbj|BAF63528.1| gloverin4 [Bombyx mori] 10/09/09 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS798027 ffbm ...

  4. EST Table: CK542390 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/31 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS917189 swh ...

  5. EST Table: CK504138 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h CK504855 swc ...

  6. EST Table: CK506233 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available byx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS917189 swc ...

  7. EST Table: DC538827 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ombyx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/09/02 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS917189 MFB- ...

  8. EST Table: CK506268 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h CK504855 swc ...

  9. EST Table: CK510389 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h CK504855 swd ...

  10. EST Table: CK555622 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/31 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS917189 swl ...

  11. EST Table: CK504533 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h CK504855 swc ...

  12. EST Table: CK506998 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h CK504855 swc ...

  13. EST Table: CK511344 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53373.1| antibacterial peptide [Bombyx mori] dbj|BAF63528.1| gloverin4 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS798027 swd ...

  14. EST Table: CK552507 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available byx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/31 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h CK504855 swl ...

  15. EST Table: FS899154 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ombyx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/09/12 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS866683 ftes ...

  16. EST Table: CK535535 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available byx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/31 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS917189 swg ...

  17. EST Table: DC535828 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ombyx mori] dbj|BAE53373.1| antibacterial peptide [Bombyx mori] dbj|BAF63528.1| gloverin4 [Bombyx mori] 10/09/01 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS798027 MFB- ...

  18. EST Table: FS796949 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ombyx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/09/09 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS917189 ffbm ...

  19. EST Table: DC536318 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ombyx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/09/02 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS917189 MFB- ...

  20. EST Table: CK511373 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h CK504855 swd ...

  1. EST Table: CK504248 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h CK504855 swc ...

  2. EST Table: CK487803 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS917189 swa ...

  3. EST Table: CK512219 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available byx mori] dbj|BAE53373.1| antibacterial peptide [Bombyx mori] dbj|BAF63528.1| gloverin4 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS798027 swd ...

  4. EST Table: CK511026 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h CK504855 swd ...

  5. EST Table: CK507414 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h CK504855 swc ...

  6. EST Table: CK505412 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53373.1| antibacterial peptide [Bombyx mori] dbj|BAF63528.1| gloverin4 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS798027 swc ...

  7. EST Table: DC537287 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ombyx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/09/02 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS917189 MFB- ...

  8. EST Table: DC535782 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ombyx mori] dbj|BAE53373.1| antibacterial peptide [Bombyx mori] dbj|BAF63528.1| gloverin4 [Bombyx mori] 10/09/01 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS917189 MFB- ...

  9. EST Table: CK553501 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available byx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/31 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h CK504855 swl ...

  10. EST Table: FS797895 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ombyx mori] dbj|BAE53373.1| antibacterial peptide [Bombyx mori] dbj|BAF63528.1| gloverin4 [Bombyx mori] 10/09/09 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS798027 ffbm ...

  11. EST Table: CK505060 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS917189 swc ...

  12. EST Table: CK512787 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/31 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h CK504855 swd ...

  13. EST Table: CK504305 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53373.1| antibacterial peptide [Bombyx mori] dbj|BAF63528.1| gloverin4 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS798027 swc ...

  14. EST Table: CK512820 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53373.1| antibacterial peptide [Bombyx mori] dbj|BAF63528.1| gloverin4 [Bombyx mori] 10/08/31 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS798027 swd ...

  15. EST Table: CK488658 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS917189 swa ...

  16. EST Table: CK504502 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h CK504855 swc ...

  17. EST Table: CK505705 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available byx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h CK504855 swc ...

  18. EST Table: CK501811 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h CK504855 swc ...

  19. EST Table: CK506015 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available yx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h CK504855 swc ...

  20. The Evolutionary Trace Analysis of Biological Clock Protein TIME-EA4 in the Silkworm Bombyx mori%家蚕生物钟蛋白TIME-EA4的进化踪迹分析

    Institute of Scientific and Technical Information of China (English)

    王文栋; 徐世清

    2014-01-01

    家蚕是卵滞育性鳞翅目模式昆虫,其滞育卵的活化与生物钟蛋白TIME-EA4密切相关.TIME-EA4具备稳定的Cu/Zn SOD活性和瞬时的ATP酶活性.目前,国内外关于家蚕TIME-EA4的研究主要集中在结构和功能上,其分子进化机制研究尚未开展.本文利用生物信息学方法对TIME-EA4进行了进化踪迹分析,结果显示TIME-EA4的重要氨基酸残基(coverage< 25%)的91.2%都用来维持与Cu/Zn SOD的序列一致性,但TIME-EA4与Cu/Zn SOD的Cu/Zn离子结合位点在氨基酸残基组成、极性、绑定原子方面都存在差异.

  1. 家蚕浓核病毒中国株非结构蛋白1(NS1)的表达%Expression of non-structural protein 1 of Bombyx mori densovirus-3

    Institute of Scientific and Technical Information of China (English)

    余蔚; 姚勤; 郭忠建; 包方; 尹慧娟; 陈克平

    2008-01-01

    利用PCR技术扩增出BmDNV-3 NS1基因,将目的基因与原核表达载体pET-30a进行连接,转化BL21 star菌并在该菌中表达,经Western blot鉴定表达的产物为BmDNV-3 NS1蛋白,纯化NS1蛋白并制备兔多克隆抗体.同时BmDNV-3 NS1基因亚克隆到杆状病毒转移载体pFastBae-HTb-eGFP中,转化BmDH10BAC感受态细胞,提取的重组Bacmid通过脂质体包埋转染家蚕BmN细胞,再以收获的重组病毒感染家蚕幼虫.家蚕BmN细胞和幼虫感染重组病毒2d后均观察到绿色荧光,经SDS-PAGE分析真核表达的产物与预测的NS1-eGFP融合蛋白大小不一致,说明NS1-eGFP融合蛋白被昆虫内源性的蛋白酶降解.降解的产物用NS1蛋白抗体进行Western blot鉴定为BmDNV-3 NS1蛋白.

  2. CRISPR/Cas9 System and Its Application in Genome Editing of Bombyx mori%CRISPR/Cas9系统及其在家蚕基因组编辑中的应用

    Institute of Scientific and Technical Information of China (English)

    周洪英; 孙波; 吴洪丽; 胡兴明; 郝瑜; 叶建美

    2016-01-01

    CRISPR/Cas9系统源于细菌获得性免疫系统,是一项可对多个物种的基因组进行高效率编辑的技术,已被广泛用于RNA靶向的基因组定向编辑.本文简要介绍了CRISPR/Cas9系统的基本作用原理和技术改进的进展,重点介绍了CRISPR/Cas9系统在家蚕功能基因研究和定向遗传改造研究中的应用及成果,并展望了该项技术在家蚕基因组编辑中应用的前景和主要的发展方向.

  3. DECISION TOOLS FOR MULBERRY THRIPS PSEUDODENDROTHRIPS MORI (NIWA, 1908) MANAGEMENT IN SERICULTURAL REGIONS; AN OVERVIEW

    Institute of Scientific and Technical Information of China (English)

    Kayvan Etebari; L. Matindoost; R.N. Singh

    2004-01-01

    Mulberry thrips Pseudodendrothrips mori (Niwa, 1908) is a major pest of mul berry trees recorded from different sericultural regions of the world. The thrips infestation affects the qualitative and quantitative characters of mulberry leaf, by direct feeding damage to leaves and the ingestion of sap, which in turn affects the silkworm cocoon crop. This is most harmful in dry climates and seasons when heavily attacked plants lose moisture heavily. Under these conditions infestation can seriously deplete yields. The seasonal population fluctuation and the degree of damage caused to the host plant are influenced by various environmental factors including climate, host-plant variety, topography, soil type, and management regimes.This article attempts to review all available documents on mulberry thrips and to discuss the practical approaches for best control of this pest.

  4. Proliferation Potential of 18-Month-Old Callus of Ananas comosus L. cv. Moris

    Directory of Open Access Journals (Sweden)

    A.E. De Silva

    2006-01-01

    Full Text Available Differential effect of plant growth regulators and additives in proliferation of 18-month-old calli of Ananas comosus L. cv. Moris were assessed in vitro. The proliferation of callus relied on the growth regulators and additives. Of the different auxins supplemented in the Murashige and Skoog (MS media, 32.22 μM α-naphthaleneacetic acid (NAA gave the highest mean fresh weight of callus (46.817 g. Medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D was inferior to NAA, while b-naphthoxy acetic acid (BNOA and p-chlorophenoxy acetic acid (4-CPA were not effective in proliferating 18-months old callus. Addition of casein hydrolysate and coconut water to NAA supplemented medium showed better proliferation and production of callus. However, in terms of callus production, NAA at 32.22 μM was economically better.

  5. An Incremental Mori-Tanaka Homogenization Scheme for Finite Strain Thermoelastoplasticity of MMCs

    Directory of Open Access Journals (Sweden)

    Helmut J. Böhm

    2010-01-01

    Full Text Available The present paper aims at computational simulations of particle reinforced Metal Matrix Composites as well as parts and specimens made thereof. An incremental Mori-Tanaka approach with isotropization of the matrix tangent operator is adopted. It is extended to account for large strains by means of co-rotational Cauchy stresses and logarithmic strains and is implemented into Finite Element Method software as constitutive material law. Periodic unit cell predictions in the finite strain regime are used to verify the analytical approach with respect to non-proportional loading scenarios and assumptions concerning finite strain localization. The response of parts made of Metal Matrix Composites is predicted by a multiscale approach based on these two micromechanical methods. Results for the mesoscopic stress and strain fields as well as the microfields are presented to demonstrate to capabilities of the developed methods.

  6. An ethanol extract of Ramulus mori improves blood circulation by inhibiting platelet aggregation.

    Science.gov (United States)

    Lee, Jiyun; Kwon, Gayeung; Park, Jieun; Kim, Jeong-Keun; Choe, Soo Young; Seo, Yoonhee; Lim, Young-Hee

    2016-07-01

    Inappropriate platelet aggregation can cause blood coagulation and thrombosis. In this study, the effect of an ethanol extract of Ramulus mori (ERM) on blood circulation was investigated. The antithrombotic activity of ERM on rat carotid arterial thrombosis was evaluated in vivo, and the effect of ERM on platelet aggregation and blood coagulation time was evaluated ex vivo. To evaluate the safety of ERM, its cytotoxicity to platelets and its effect on tail bleeding time were assessed; ERM was not toxic to rat platelets and did not prolong bleeding time. Moreover, administering ERM to rats had a significant preventive effect on carotid arterial thrombosis in vivo, and significantly inhibited adenosine diphosphate- and collagen-induced platelet aggregation ex vivo, whereas it did not prolong coagulation periods, such as prothrombin time and activated partial thromboplastin time. The results suggest that ERM is effective in improving blood circulation via antiplatelet activity rather than anticoagulation activity. PMID:26967156

  7. EST Table: NM_001114987 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available NM_001114987 Dopr1 11/12/09 GO hit GO:0004952(dopamine ... receptor activity)|GO:0007186(G-protein c ... embrane) 10/09/29 100 %/408 aa ref|NP_001108459.1| dopamine ... receptor-1 [Bombyx mori] dbj|BAF98647.1| dopamine ... receptor-1 [Bombyx mori] dbj|BAG14260.1| dopamine ... receptor 1 [Bombyx mori] 10/09/13 72 %/368 aa FBpp ...

  8. EST Table: NM_001044219 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available NM_001044219 glv4 10/09/29 100 %/171 aa ref|NP_001037684.1| gloverin 4 [Bombyx mori] dbj|BAE53373.1| antibac...terial peptide [Bombyx mori] dbj|BAF63528.1| gloverin4 [Bombyx mori] 10/09/13 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS798027 ...

  9. EST Table: NM_001044218 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available NM_001044218 glv2 10/09/29 100 %/173 aa ref|NP_001037683.1| gloverin 2 [Bombyx mori] dbj|BAE53372.1| antibac...terial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/09/13 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS917189 ...

  10. EST Table: FS917049 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In A...u-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Sclerosis pdb|...3L9E|C Chain C, Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implication...s Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral

  11. EST Table: FS784229 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In A...u-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Sclerosis pdb|...3L9E|C Chain C, Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implication...s Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral

  12. EST Table: FS828896 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In A...u-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Sclerosis pdb|...3L9E|C Chain C, Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implication...s Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral

  13. EST Table: FY018191 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophi...nt CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Sclerosis pdb|3L9E|C Ch...ain C, Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implication... And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Sclerosi

  14. EST Table: FS824641 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In A...u-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Sclerosis pdb|...3L9E|C Chain C, Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implication...s Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral

  15. EST Table: FS756290 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In A...u-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Sclerosis pdb|...3L9E|C Chain C, Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implication...s Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral

  16. EST Table: FS765772 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In A...u-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Sclerosis pdb|...3L9E|C Chain C, Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implication...s Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral

  17. EST Table: FS917301 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Scl...OM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Sclerosis pdb|3L9E|D Chain D, Crysta...l Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrop...ications In Amyotrophic Lateral Sclerosis pdb|3L9E|B Chain B, Crystal Structures Of...|A Chain A, Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Impl

  18. EST Table: FS728648 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In A...u-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Sclerosis pdb|...3L9E|C Chain C, Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implication...s Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral

  19. EST Table: FS886767 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In A...u-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Sclerosis pdb|...3L9E|C Chain C, Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implication...s Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral

  20. EST Table: FS877359 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In A...u-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Sclerosis pdb|...3L9E|C Chain C, Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implication...s Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral