WorldWideScience

Sample records for bombyx mori revealed

  1. 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 ...

  2. 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.

  3. 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

  4. Nucleotide Diversity and Selection Signature in the Domesticated Silkworm, Bombyx mori, and Wild Silkworm, Bombyx mandarina

    OpenAIRE

    Guo, Yi; Shen, Yi-Hong; Sun, Wei; Kishino, Hirohisa; Xiang, Zhong-Huai; Zhang, Ze

    2011-01-01

    To investigate the patterns of nucleotide diversity in domesticated silkworm, Bombyx mori L. (Lepidoptera: Bombycidae) and its wild relative, Chinese wild silkworm, Bombyx mandarina Moore, we sequenced nine nuclear genes. Neutrality test and coalescent simulation for these genes were performed to look at bottleneck intensity and selection signature; linkage disequilibrium (LD) within and between loci was employed to investigate allele association. As a result, B. mori lost 33–49% of nucleotid...

  5. 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.

  6. 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.

  7. 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...

  8. 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 ...

  9. 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 ...

  10. Nucleotide diversity and selection signature in the domesticated silkworm, Bombyx mori, and wild silkworm, Bombyx mandarina.

    Science.gov (United States)

    Guo, Yi; Shen, Yi-Hong; Sun, Wei; Kishino, Hirohisa; Xiang, Zhong-Huai; Zhang, Ze

    2011-01-01

    To investigate the patterns of nucleotide diversity in domesticated silkworm, Bombyx mori L. (Lepidoptera: Bombycidae) and its wild relative, Chinese wild silkworm, Bombyx mandarina Moore, we sequenced nine nuclear genes. Neutrality test and coalescent simulation for these genes were performed to look at bottleneck intensity and selection signature; linkage disequilibrium (LD) within and between loci was employed to investigate allele association. As a result, B. mori lost 33-49% of nucleotide diversity relative to wild silkworm, which is similar to the loss levels found in major cultivated crops. Diversity of B. mori is significantly lower than that of B. mandarina measured as π(total) (0.01166 vs. 0.1741) or θ(W)(0.01124 vs. 0.02206). Bottleneck intensity of domesticated silkworm is 1.5 (in terms of k = N(b) /d, N(b) -bottleneck population size; d-bottleneck duration) with different durations. Gene DefA showed signature of artificial selection by all analysis methods and might experience strong artificial selection in B. mori during domestication. For nine loci, both curves of LD decay rapidly within 200 bp and drop slowly when distance is > 200 bp, although that of B. mori decays slower than B. mandarina at loci investigated. However, LD could not be estimated at DefA in B. mori and at ER in both silkworms. Elevated LD observed in B. mori may be indicator of selection and demographic events. PMID:22239062

  11. 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.

  12. Transposable-element associated small RNAs in Bombyx mori genome.

    Directory of Open Access Journals (Sweden)

    Yimei Cai

    Full Text Available Small RNAs are a group of regulatory RNA molecules that control gene expression at transcriptional or post-transcriptional levels among eukaryotes. The silkworm, Bombyx mori L., genome harbors abundant repetitive sequences derived from families of retrotransposons and transposons, which together constitute almost half of the genome space and provide ample resource for biogenesis of the three major small RNA families. We systematically discovered transposable-element (TE-associated small RNAs in B. mori genome based on a deep RNA-sequencing strategy and the effort yielded 182, 788 and 4,990 TE-associated small RNAs in the miRNA, siRNA and piRNA species, respectively. Our analysis suggested that the three small RNA species preferentially associate with different TEs to create sequence and functional diversity, and we also show evidence that a Bombyx non-LTR retrotransposon, bm1645, alone contributes to the generation of TE-associated small RNAs in a very significant way. The fact that bm1645-associated small RNAs partially overlap with each other implies a possibility that this element may be modulated by different mechanisms to generate different products with diverse functions. Taken together, these discoveries expand the small RNA pool in B. mori genome and lead to new knowledge on the diversity and functional significance of TE-associated small RNAs.

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

    Directory of Open Access Journals (Sweden)

    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.

  14. 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.

  15. 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

  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. Structural study of Bombyx mori silk fibroin during processing for regeneration

    Science.gov (United States)

    Ha, Sung-Won

    Bombyx mori silk fibroin has excellent mechanical properties combined with flexibility, tissue compatibility, and high oxygen permeability in the wet condition. This important material should be dissolved and regenerated to be utilized as useful forms such as gel, film, fiber, powder, or non-woven. However, it has long been a problem that the regenerated fibroin materials show poor mechanical properties and brittleness. These problems were technically solved by improving a fiber processing method reported here. The regenerated fibroin fibers showed much better mechanical properties compared to the original silk fibers. This improved technique for the fiber processing of Bombyx mori silk fibroin may be used as a model system for other semi-crystalline fiber forming proteins, becoming available through biotechnology. The physical and chemical properties of the regenerated fibers were characterized by SinTechRTM tensile testing, X-ray diffraction, solid state 13C NMR spectroscopy, and SEM. Unlike synthetic polymers, the molecular weight distribution of Bombyx mori silk fibroin is mono-disperse because silk fibroin is synthesized from DNA template. Genetic studies have revealed the entire amino acid sequence of Bombyx mori silk fibroin. It is known that the crystalline silk II structure is composed of hexa-amino acid sequences, GAGAGS. However, in the amino acid sequence of Bombyx mori silk fibroin heavy chain, there are present 11 chemically irregular but evolutionarily conserved sequences with about 31 amino acid residues (irregular GT˜GT sequences). The structure and role of these irregular sequences have remained unknown. One of the most frequently appearing irregular sequences was synthesized by a peptide synthesizer. The three-dimensional structure of this irregular silk peptide was studied by the high resolution two-dimensional NMR technique. The three-dimensional structure of this peptide shows that it makes a turn or loop structure (distorted O shape), which

  18. Molecular Systematic Studies on Chinese Mandarina Silkworm (Bombyx mandarina M. ) and Domestic Silkworm (Bombyx mori L.)

    Institute of Scientific and Technical Information of China (English)

    LU Cheng; YU Hong-shi; XIANG Zhong-huai

    2002-01-01

    Molecular systematic studies on mandarina silkworm (Bombyx mandarina M. ) in 11 regions in China and 25 representative strains of domestic silkworm (Bombyx mori L. ) were conducted using molecular biology techniques. Results obtained from the analysis of DNA polymorphism and clustering of all the silkworm samples provide new evidence for the view that the domestic silkworm originated from the Chinese mandarina silkworm. On the basis of literature reviewing, a new hypothesis on the origin of the domestic silkworm was put forward. It was thought that the domestic silkworm was most probably domesticated from the Chinese mandarina silkworm of different ecotypes including monovoltinism, bivoitinism and multivoitinism; and that the domestic silkworm had the genetic background of monovoltinism, bivoltinism and multivoltinism at the very beginning of the domestication. The current strains of the domestic silkworm of different voltinism are the evolutionary results of thousands of years of rearing and artificial selections.

  19. 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

  20. Investigation of Natural Bombyx mori Silk Fibroin Proteins Using INS

    Science.gov (United States)

    Crain, Christopher; Strange, Nicholas; Larese, J. Z.

    The mechanical properties of many protein comprised biomaterials are a direct reflection of non-covalent (i.e. weak) interacting ions such as F-actin in muscles, tubulin in the cytoskeleton of cells, viral capsids, and silk. Porter and Vollrath underscored the two main factors that are critical for understanding the high mechanical strength of silks: the nanoscale semi-crystalline folding structure, which gives it exceptional toughness and strength, and the degree of hydration of the disordered fraction, which acts to modify these properties. Understanding and controlling these two principal factors are the key to the functionality of protein elastomers, and render silk an ideal model protein for (bio)material design. We will describe our investigation of electrospun silk of the Bombyx mori (silk worm), using Inelastic Neutron Scattering (INS). These techniques were used to investigate the microscopic dynamics of the dry and hydrated protein.

  1. 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.

  2. 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

  3. 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

  4. 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.

  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. 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.

  7. Identification of C-type lectin-domain proteins (CTLDPs) in silkworm Bombyx mori.

    Science.gov (United States)

    Rao, Xiang-Jun; Shahzad, Toufeeq; Liu, Su; Wu, Peng; He, Yan-Ting; Sun, Wei-Jia; Fan, Xiang-Yun; Yang, Yun-Fan; Shi, Qiao; Yu, Xiao-Qiang

    2015-12-01

    C-type lectins (CTLs) represent a large family of proteins that can bind carbohydrate moieties normally in a calcium-dependent manner. CTLs play important roles in mediating cell adhesion and the recognition of pathogens in the immune system. In the present study, we have identified 23 CTL genes in domestic silkworm Bombyx mori. CTL-domain proteins (CTLDPs) are classified into three groups based on the number of carbohydrate-recognition domains (CRDs) and the domain architectures. These include twelve CTL-S (Single-CRD), six immulectins (Dual-CRD) and five CTL-X (CRD with other domains). We studied their phylogenetic features, analyzed the conserved residues, predicted tertiary structures, and examined the tissue expression profile and immune inducibility. Through bioinformatics analysis, we have putatively identified ten secretory and two cytoplasmic CTL-S; four secretory and two cytoplasmic immulectins; one secretory, one cytoplasmic and three transmembrane forms of CTL-X. Most B. mori CTLDPs form monophyletic groups with orthologs from Lepidoptera, Diptera, Coleoptera and Hymenoptera species. Immulectins of B. mori and Manduca sexta evolved from common ancestor genes perhaps due to gene duplication events of CTL-S ancestor genes. Homology modeling revealed that the overall structures of B. mori CTL domains are analogous to those of humans with a variable loop region. We examined the expression profile of CTLDP genes in naïve and immune-stimulated tissues. The expression and induction of CTLDP genes were related to the tissues and microorganisms. Together, our gene identification, sequence comparison, phylogenetic analysis, homology modeling and expression analysis laid a good foundation for the further studies of B. mori CTLDPs and comparative genomics. PMID:26187302

  8. Genome-wide identification, characterization of sugar transporter genes in the silkworm Bombyx mori and role in Bombyx mori nucleopolyhedrovirus (BmNPV) infection.

    Science.gov (United States)

    Govindaraj, Lekha; Gupta, Tania; Esvaran, Vijaya Gowri; Awasthi, Arvind Kumar; Ponnuvel, Kangayam M

    2016-04-01

    Sugar transporters play an essential role in controlling carbohydrate transport and are responsible for mediating the movement of sugars into cells. These genes exist as large multigene families within the insect genome. In insects, sugar transporters not only have a role in sugar transport, but may also act as receptors for virus entry. Genome-wide annotation of silkworm Bombyx mori (B. mori) revealed 100 putative sugar transporter (BmST) genes exists as a large multigene family and were classified into 11 sub families, through phylogenetic analysis. Chromosomes 27, 26 and 20 were found to possess the highest number of BmST paralogous genes, harboring 22, 7 and 6 genes, respectively. These genes occurred in clusters exhibiting the phenomenon of tandem gene duplication. The ovary, silk gland, hemocytes, midgut and malphigian tubules were the different tissues/cells enriched with BmST gene expression. The BmST gene BGIBMGA001498 had maximum EST transcripts of 134 and expressed exclusively in the malphigian tubule. The expression of EST transcripts of the BmST clustered genes on chromosome 27 was distributed in various tissues like testis, ovary, silk gland, malphigian tubule, maxillary galea, prothoracic gland, epidermis, fat body and midgut. Three sugar transporter genes (BmST) were constitutively expressed in the susceptible race and were down regulated upon BmNPV infection at 12h post infection (hpi). The expression pattern of these three genes was validated through real-time PCR in the midgut tissues at different time intervals from 0 to 30hpi. In the susceptible B. mori race, expression of sugar transporter genes was constitutively expressed making the host succumb to viral infection.

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

    Directory of Open Access Journals (Sweden)

    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

  10. 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...

  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. Characterization of Cryptopygus antarcticus endo-β-1,4-glucanase from Bombyx mori expression systems.

    Science.gov (United States)

    Hong, Sun Mee; Sung, Ho Sun; Kang, Mee Hye; Kim, Choong-Gon; Lee, Youn-Ho; Kim, Dae-Jung; Lee, Jae Man; Kusakabe, Takahiro

    2014-10-01

    Endo-β-1,4-glucanase (CaCel) from Antarctic springtail, Cryptopygus antarcticus, a cellulase with high activity at low temperature, shows potential industrial use. To obtain sufficient active cellulase for characterization, CaCel gene was expressed in Bombyx mori-baculovirus expression systems. Recombinant CaCel (rCaCel) has been expressed in Escherichia coli (Ec-CaCel) at temperatures below 10°C, but the expression yield was low. Here, rCaCel with a silkworm secretion signal (Bm-CaCel) was successfully expressed and secreted into pupal hemolymph and purified to near 90% purity by Ni-affinity chromatography. The yield and specific activity of rCaCel purified from B. mori were estimated at 31 mg/l and 43.2 U/mg, respectively, which is significantly higher than the CaCel yield obtained from E. coli (0.46 mg/l and 35.8 U/mg). The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. Thermostability of Bm-CaCel from B. mori at 60°C was higher than that from E. coli, probably due to glycosylation. PMID:24848382

  13. Three-dimensional structure of a Bombyx mori Omega-class glutathione transferase.

    Science.gov (United States)

    Yamamoto, Kohji; Suzuki, Mamoru; Higashiura, Akifumi; Nakagawa, Atsushi

    2013-09-01

    Glutathione transferases (GSTs) are major phase II detoxification enzymes that play central roles in the defense against various environmental toxicants as well as oxidative stress. Here we report the crystal structure of an Omega-class glutathione transferase of Bombyx mori, bmGSTO, to gain insight into its catalytic mechanism. The structure of bmGSTO complexed with glutathione determined at a resolution of 2.5Å reveals that it exists as a dimer and is structurally similar to Omega-class GSTs with respect to its secondary and tertiary structures. Analysis of a complex between bmGSTO and glutathione showed that bound glutathione was localized to the glutathione-binding site (G-site). Site-directed mutagenesis of bmGSTO mutants indicated that amino acid residues Leu62, Lys65, Lys77, Val78, Glu91 and Ser92 in the G-site contribute to catalytic activity.

  14. 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.

  15. 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.

  16. 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

  17. 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.

  18. Large scale full-length cDNA sequencing reveals a unique genomic landscape in a lepidopteran model insect, Bombyx mori.

    Science.gov (United States)

    Suetsugu, Yoshitaka; Futahashi, Ryo; Kanamori, Hiroyuki; Kadono-Okuda, Keiko; Sasanuma, Shun-ichi; Narukawa, Junko; Ajimura, Masahiro; Jouraku, Akiya; Namiki, Nobukazu; Shimomura, Michihiko; Sezutsu, Hideki; Osanai-Futahashi, Mizuko; Suzuki, Masataka G; Daimon, Takaaki; Shinoda, Tetsuro; Taniai, Kiyoko; Asaoka, Kiyoshi; Niwa, Ryusuke; Kawaoka, Shinpei; Katsuma, Susumu; Tamura, Toshiki; Noda, Hiroaki; Kasahara, Masahiro; Sugano, Sumio; Suzuki, Yutaka; Fujiwara, Haruhiko; Kataoka, Hiroshi; Arunkumar, Kallare P; Tomar, Archana; Nagaraju, Javaregowda; Goldsmith, Marian R; Feng, Qili; Xia, Qingyou; Yamamoto, Kimiko; Shimada, Toru; Mita, Kazuei

    2013-09-01

    The establishment of a complete genomic sequence of silkworm, the model species of Lepidoptera, laid a foundation for its functional genomics. A more complete annotation of the genome will benefit functional and comparative studies and accelerate extensive industrial applications for this insect. To realize these goals, we embarked upon a large-scale full-length cDNA collection from 21 full-length cDNA libraries derived from 14 tissues of the domesticated silkworm and performed full sequencing by primer walking for 11,104 full-length cDNAs. The large average intron size was 1904 bp, resulting from a high accumulation of transposons. Using gene models predicted by GLEAN and published mRNAs, we identified 16,823 gene loci on the silkworm genome assembly. Orthology analysis of 153 species, including 11 insects, revealed that among three Lepidoptera including Monarch and Heliconius butterflies, the 403 largest silkworm-specific genes were composed mainly of protective immunity, hormone-related, and characteristic structural proteins. Analysis of testis-/ovary-specific genes revealed distinctive features of sexual dimorphism, including depletion of ovary-specific genes on the Z chromosome in contrast to an enrichment of testis-specific genes. More than 40% of genes expressed in specific tissues mapped in tissue-specific chromosomal clusters. The newly obtained FL-cDNA sequences enabled us to annotate the genome of this lepidopteran model insect more accurately, enhancing genomic and functional studies of Lepidoptera and comparative analyses with other insect orders, and yielding new insights into the evolution and organization of lepidopteran-specific genes.

  19. Phosphatase activity of Bombyx mori nucleopolyhedrovirus PTP is dispensable for enhanced locomotory activity in B. mori larvae.

    Science.gov (United States)

    Katsuma, Susumu

    2015-11-01

    Baculovirus-induced enhanced locomotory activity (ELA) is not induced in caterpillars infected with a mutant Bombyx mori nucleopolyhedrovirus (BmNPV) or Autographa californica multiple nucleopolyhedrovirus (AcMNPV) lacking a functional protein tyrosine phosphatase gene (ptp). Previous studies suggest that the PTP proteins from BmNPV and AcMNPV act in different ways to induce ELA, i.e., BmNPV PTP is utilized as a virion structural component, whereas AcMNPV PTP requires its phosphatase activity. Here, I generated and characterized two new BmNPV mutants expressing enzymatically inactive PTP proteins and confirmed that the phosphatase activity of PTP is not required for ELA induction in BmNPV-infected B. mori larvae.

  20. Effect of Wheat Flour Noodles with Bombyx mori Powder on Glycemic Response in Healthy Subjects

    Science.gov (United States)

    Suk, Wanhee; Kim, JiEun; Kim, Do-Yeon; Lim, Hyunjung; Choue, Ryowon

    2016-01-01

    Recent trial results suggest that the consumption of a low glycemic index (GI) diet is beneficial in the prevention of high blood glucose levels. Identifying active hypoglycemic substances in ordinary foods could be a significant benefit to the management of blood glucose. It has been hypothesized that noodles with Bombyx mori powder are a low GI food. We evaluated GI and changes in postprandial glucose levels following consumption of those noodles and compared them with those following consumption of plain wheat flour noodles (control) and glucose (reference) in healthy subjects. Thirteen males (age: 34.2±4.5 years, body mass index: 23.2±1.1 kg/m2) consumed 75 g carbohydrate portions of glucose and the 2 kinds of noodle after an overnight fast. Capillary blood was measured at time 0 (fasting), 15, 30, 45, 60, 90, 120, and 180 min from the start of each food intake. The GI values were calculated by taking the ratio of the incremental area under the blood glucose response curve (IAUC) for the noodles and glucose. There was a significant difference in postprandial glucose concentrations at 30 and 45 min between the control noodles and the noodles with Bombyx mori powder: the IAUC and GI for the noodles with Bombyx mori powder were significantly lower than those for glucose and plain wheat flour noodles. The wheat flour noodles with Bombyx mori powder could help prevent an increase in postprandial glucose response and possibly provide an alternative to other carbohydrate staple foods for glycemic management. PMID:27752491

  1. Structural insight into the active site of a Bombyx mori unclassified glutathione transferase.

    Science.gov (United States)

    Hossain, Md Tofazzal; Yamamoto, Kohji

    2015-01-01

    Glutathione transferases (GSTs) are major detoxification enzymes that play central roles in the defense against various environmental toxicants as well as oxidative stress. Here, we identify amino acid residues of an unclassified GST from Bombyx mori, bmGSTu-interacting glutathione (GSH). Site-directed mutagenesis of bmGSTu mutants indicated that amino acid residues Asp103, Ser162, and Ser166 contribute to catalytic activity.

  2. 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 ...

  3. Cloning of TRP Gene of Bombyx mori%家蚕TPR基因的克隆

    Institute of Scientific and Technical Information of China (English)

    牛伟涛

    2011-01-01

    [ Objective ] The aim was to clone Bombyx mori TRP gene. [ Method ] The total RNA of Bombyx mori was extracted during its pupal period with Trizol method, and then the TRP gene was cloned by RT-PCR. [ Result] The full-length cDNA of TRP gene was successfully obtained, and the ORF fragment (858 bp) of TRP gene was cloned by PCR. [Conclusion] TRP gene of Bombyx mori was cloned for the first time, which could provide solid basis for the study on its function.%[目的]克隆家蚕TPR基因,为研究该基因功能提供依据.[方法]利用Trizol法提取家蚕蛹期总RNA,再用RT-PCR方法对家蚕TPR基因进行克隆.[结果]成功得到家蚕TPR基因全cDNA序列,并用PCR扩增得到了TPR基因的ORF,全长858 bp.[结论]首次对家蚕TPR家族基因进行了克隆,为今后对该基因功能研究奠定了坚实的基础.

  4. 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

  5. Anatomical and functional analysis of domestication effects on the olfactory system of the silkmoth Bombyx mori.

    Science.gov (United States)

    Bisch-Knaden, Sonja; Daimon, Takaaki; Shimada, Toru; Hansson, Bill S; Sachse, Silke

    2014-01-01

    The silkmoth Bombyx mori is the main producer of silk worldwide and has furthermore become a model organism in biological research, especially concerning chemical communication. However, the impact domestication might have had on the silkmoth's olfactory sense has not yet been investigated. Here, we show that the pheromone detection system in B. mori males when compared with their wild ancestors Bombyx mandarina seems to have been preserved, while the perception of environmental odorants in both sexes of domesticated silkmoths has been degraded. In females, this physiological impairment was mirrored by a clear reduction in olfactory sensillum numbers. Neurophysiological experiments with hybrids between wild and domesticated silkmoths suggest that the female W sex chromosome, so far known to have the sole function of determining femaleness, might be involved in the detection of environmental odorants. Moreover, the coding of odorants in the brain, which is usually similar among closely related moths, differs strikingly between B. mori and B. mandarina females. These results indicate that domestication has had a strong impact on odour detection and processing in the olfactory model species B. mori.

  6. 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

  7. 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.

  8. 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

  9. 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.

  10. Paralogous gene conversion, allelic divergence of attacin genes and its expression profile in response to BmNPV infection in silkworm Bombyx mori

    Directory of Open Access Journals (Sweden)

    G Lekha

    2015-08-01

    Full Text Available The genomic organization, structure and polymorphism of attacin gene within the mulberry silkworm Bombyx mori strains have been analyzed. Genomic contig (AADK01007556 of B. mori attacin gene contains locus with two transcribed basic attacin genes, which were designated as attacin I and attacin II. Survey of the naturally occurring genetic variation in different strains of silkworm B. mori at the promoter and coding regions of two attacin genes revealed high levels of silent nucleotide variations (1- 4 % per nucleotide heterozygosity without polymorphism at the amino acid level (nonSynonymous substitution. We also investigated variations in gene expression of attacin I and attacin II in silkworm B. mori infected with nucleopolyhedrovirus (BmNPV. Two B. mori strains, Sarupat, CSR-2 which were resistant and susceptible to BmNPV infection respectively were used in this study. Expression profiles of B. mori genes were analyzed using microarray technique and results revealed that the immune response genes including attacin were selectively up regulated in virus invaded midguts of both races. Microarray data and real-time qPCR results revealed that attacin I gene was significantly up-regulated in the midgut of Sarupat following BmNPV infection, indicating its specific role in the anti-viral response. Our results imply that these up-regulated attacin genes were not only involved in anti-bacterial mechanism, but are also involved in B. mori immune response against BmNPV infection.

  11. Molecular Markers for Biomass Traits: Association, Interaction and Genetic Divergence in Silkworm Bombyx mori

    Directory of Open Access Journals (Sweden)

    Appukuttannair R Pradeep

    2007-01-01

    Full Text Available Improvement of high yielding, disease resistant silkworm strains became imminent to increase production of silk, which is a major revenue earner for sericulturists. Since environment interacts with phenotype, conventional breeding did not result in commendable yield improvement in synthetic strains of silkworm, Bombyx mori. Identification of DNA markers associated with different economically important biomass traits and its introgression could assist molecular breeding and expression of stabilized high yielding characters, but genetic basis of most quantitative traits in silkworm is poorly understood due to its polygenic control. Correlation analysis (R = 0.9 revealed significant interrelation among biomass traits viz., larval duration (TLD, larval weight (LWT, cocoon weight (CWT, shell weight (SWT, shell ratio (SR and floss content. PCR using inter simple sequence repeat (ISSR primers revealed 92% polymorphism among 14 tropical and temperate strains of B. mori, with average diversity index of 0.747. Stepwise multiple regression analysis (MRA selected 35 ISSR markers positively or negatively correlated with different biomass traits, illustrated polygenic control. ISSR marker 830.81050bp was significantly associated with LWT, CWT, SWT, SR and fl oss content, indicated its pleiotropic role. Two ISSR markers, 835.51950bp and 825.9710bp showed significant association with floss content and TLD. These markers were segregated in F2 generation and Chi-square test confirmed (χ2 = ~45; P < 0.05 its genetic contribution to the associated biomass traits. Strains, with both positively and negatively correlated markers, had intermediate mean value for biomass traits (eg. SWT = 0.17 ± 0.014 g in GNM and Moria indicated interaction of loci in natural populations. Low yielding Indian strains grouped together by Hierarchical clustering. Chinese and Japanese strains were distributed in the periphery of ALSCAL matrix indicated convergence of genetic characters

  12. Molecular characterization, tissue distribution, and immune reaction expression of karyopherins in the domestic silkworm (Bombyx mori).

    Science.gov (United States)

    Li, J; Wang, L; Qian, C; Zhang, C F; Dai, L S; Liu, Q N; Wei, G Q; Sun, Y; Liu, D R; Zhu, B J; Liu, C L

    2015-01-01

    Karyopherins, including alpha and beta types, are transport proteins in the eukaryotic cell that carry cargoes across nuclear pore complexes into or out of the nucleus. In this study, full open reading frames of one beta and three alpha types of karyopherin were cloned from cDNA of the domestic silkworm (Bombyx mori). The one beta and three alpha types' open reading frames were 2661, 1563, 1515, and 1551 base pairs long, respectively, and coded 886, 520, 504, and 516 amino acids, respectively. The alphas all had one importin-beta-binding (IBB) domain, and eight, four, or seven armadillo/beta-catenin-like repeats. The beta had 19 HEAT repeat domains, which constructed one importin-beta-N-terminal domain and one IBB domain. The recombinant proteins were expressed in Escherichia coli cells. The molecular weight of the beta type was approximately 100 kDa, and the alphas weighed approximately 60 kDa. Phylogenic tree construction revealed that the alphas could be classified into three known karyopherin-alpha subfamilies. We detected mRNA of the four karyopherins in normal 3rd day of 5th instar larvae, and in larvae injected with Gram-positive bacteria, Gram-negative bacteria, viruses, and fungi using real-time fluorescence quantitative reverse transcriptase-polymerase chain reaction, and found that the four karyopherins were widely distributed, but their expression levels were related to tissues type, the microbe injected, and the time point. PMID:26535618

  13. 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.

  14. 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

  15. 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

  16. 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.

  17. Drosophila intersex orthologue in the silkworm, Bombyx mori and related species.

    Science.gov (United States)

    Arunkumar, K P; Nagaraju, J

    2011-01-01

    Intersex (ix), a gene required for female sexual development in Drosophila, acts in concert with doublesex (dsx) at the end of the sex determination pathway. In the present study a homologue of ix was identified in Bombyx mori. Expression analysis of this gene by RT-PCR and RNase protection assay revealed a diagnostic alternative splice form present only in testis, whereas the most common splice form was found to express in all other tissues from early embryonic developmental stages. The present study provides evidence for the presence of an alternative splice form of ix in three species of silkmoths examined. Taken together with the results of an earlier study on ix in piralid moth, Maruca vitrata (Cavaliere et al. 2009), the present study suggests that the testis-specific splice form may be a characteristic feature of lepidopterans. Though ix lacks a conserved splicing pattern it appears to have retained its functional conservation in terminal sexual differentiation. We speculate that the presence of an additional splice form, perhaps encoding non-functional protein only in testis, may prevent the feminizing effects exerted by the functional IX protein.

  18. Bombyx mori E26 transformation-specific 2 (BmEts2), an Ets family protein, represses Bombyx mori Rels (BmRels)-mediated promoter activation of antimicrobial peptide genes in the silkworm Bombyx mori.

    Science.gov (United States)

    Tanaka, H; Sagisaka, A; Suzuki, N; Yamakawa, M

    2016-10-01

    E26 transformation-specific (Ets) family transcription factors are known to play roles in various biological phenomena, including immunity, in vertebrates. However, the mechanisms by which Ets proteins contribute to immunity in invertebrates remain poorly understood. In this study, we identified a cDNA encoding BmEts2, which is a putative orthologue of Drosophila Yan and human translocation-ets-leukemia/Ets-variant gene 6, from the silkworm Bombyx mori. Expression of the BmEts2 gene was significantly increased in the fat bodies of silkworm larvae in response to injection with Escherichia coli and Staphylococcus aureus. BmEts2 overexpression dramatically repressed B. mori Rels (BmRels)-mediated promoter activation of antimicrobial peptide genes in silkworm cells. Conversely, gene knockdown of BmEts2 significantly enhanced BmRels activity. In addition, two κB sites located on the 5' upstream region of cecropin B1 were found to be involved in the repression of BmRels-mediated promoter activation. Protein-competition analysis further demonstrated that BmEts2 competitively inhibited binding of BmRels to κB sites. Overall, BmEts2 acts as a repressor of BmRels-mediated transactivation of antimicrobial protein genes by inhibiting the binding of BmRels to κB sites.

  19. 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.

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

    Science.gov (United States)

    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

  1. 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.

  2. Conformational Transformation Exhibited by the Peptide Extracted from Crystalline Region of Bombyx mori Silk Fibroin in Solid and Solution States

    Institute of Scientific and Technical Information of China (English)

    YAO Ju-Ming; ZHANG Guo-Qing; LEI Cai-Hong

    2006-01-01

    The conformational transformation of a 30-residue peptide H(Ala-Gly-Ser-Gly-Ala-Gly)5OH, i.e., (AGSGAG)5,extracted from highly crystalline region of Bombyx mori (B. mori) silk fibroin was described by using the high resolution solid state 13C NMR, and CD spectroscopies. Based on the conformation-dependent 13C NMR chemical shifts of the Ala, Gly and Ser residues and the line-shape analysis of the conformation sensitive Ala Cβ resonance,the peptide revealed a strong preference for silk Ⅱ structural form, i.e., an antiparallel β-sheet structure (φ=-140±20° and ψ= 135±20°) in solid state. On the contrary, the CD spectra of this peptide in the two non-native hexafluorinated fibre spinning solvents, hexafluoroisopropanol (HFIP) and hexafluoroacetone (HFA), exhibited the existence of an unusual tightly-folded conformation resembling 310-helix (φ=-60±20° and ψ=- 30 ± 20°), as judged from the R ratio of [θ]222/[θ]203 in HFIP solution, whereas a dynamically averaged unordered structure in HFA. Taken together, the information inclined to hypothesis that the primary structure of the highly crystalline regions of B. mori silk fibroin may be easily accessible to the large conformational changes, which in turn may be critical for facilitating the structural transformation from unprocessed silk fibroin (silk Ⅰ form) to processed silk fiber (silk Ⅱ form).

  3. 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.

  4. 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.

  5. Targeting of human aFGF gene into silkworm, Bombyx mori L.Through homologous recombination

    Institute of Scientific and Technical Information of China (English)

    吴小锋; 曹翠平

    2004-01-01

    The long-arm and short-arm genes of fibroin light chain (L-chain) of silkworm, Bombyx Mori L., and the gene of human acidic fibroblast growth factor were cloned respectively and subsequently inserted into a transfer vector pVL1392 used as a tool to target the L-chain region of the silkworm genome. Genomic DNA from their offsprings was extracted and the expected targeting was detected using polymerase chain reaction and DNA sequencing, as well as protein analysis. The results showed that positive events occurred and that the FGF gene was integrated into the L-chain locus through homologous recombination.

  6. 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...

  7. Structural and thermal properties of γ – irradiated Bombyx mori silk fibroin films

    Energy Technology Data Exchange (ETDEWEB)

    Madhukumar, R.; Asha, S.; Rao, B. Lakshmeesha; Shivananda, C. S.; Harish, K. V.; Sangappa, E-mail: syhalabhavi@yahoo.co.in [Department of Studies in Physics, Mangalore University, Mangalagangotri, Mangalore - 574199 (India); Sarojini, B. K. [Department of Studies in Chemistry, Mangalore University, Mangalagangotri, Mangalore - 574199 (India); Somashekar, R. [Department of Studies in Physics, University of Mysore, Manasagangotri, Mysore - 570006 (India)

    2015-06-24

    The gamma radiation-induced change in structural and thermal properties of Bombyx mori silk fibroin films were investigated and have been correlated with the applied radiation doses. Irradiation of samples were carried out in dry air at room temperature using Co-60 source, and radiation doses are in the range of 0 - 300 kGy. Structural and thermal properties of the irradiated silk films were studied using X-ray diffraction (XRD), Differential Scanning Calorimetry (DSC) and Thermogravimetric analysis (TGA) and compared with unirradiated sample. Interesting results are discussed in this report.

  8. 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.

  9. 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.

  10. 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

  11. 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

  12. Characterization of anti-CD20 monoclonal antibody produced by transgenic silkworms (Bombyx mori)

    Science.gov (United States)

    Tada, Minoru; Tatematsu, Ken-Ichiro; Ishii-Watabe, Akiko; Harazono, Akira; Takakura, Daisuke; Hashii, Noritaka; Sezutsu, Hideki; Kawasaki, Nana

    2015-01-01

    In response to the successful use of monoclonal antibodies (mAbs) in the treatment of various diseases, systems for expressing recombinant mAbs using transgenic animals or plants have been widely developed. The silkworm (Bombyx mori) is a highly domesticated insect that has recently been used for the production of recombinant proteins. Because of their cost-effective breeding and relatively easy production scale-up, transgenic silkworms show great promise as a novel production system for mAbs. In this study, we established a transgenic silkworm stably expressing a human-mouse chimeric anti-CD20 mAb having the same amino acid sequence as rituximab, and compared its characteristics with rituximab produced by Chinese hamster ovary (CHO) cells (MabThera®). The anti-CD20 mAb produced in the transgenic silkworm showed a similar antigen-binding property, but stronger antibody-dependent cell-mediated cytotoxicity (ADCC) and weaker complement-dependent cytotoxicity (CDC) compared to MabThera. Post-translational modification analysis was performed by peptide mapping using liquid chromatography/mass spectrometry. There was a significant difference in the N-glycosylation profile between the CHO− and the silkworm-derived mAbs, but not in other post-translational modifications including oxidation and deamidation. The mass spectra of the N-glycosylated peptide revealed that the observed biological properties were attributable to the characteristic N-glycan structures of the anti-CD20 mAbs produced in the transgenic silkworms, i.e., the lack of the core-fucose and galactose at the non-reducing terminal. These results suggest that the transgenic silkworm may be a promising expression system for the tumor-targeting mAbs with higher ADCC activity. PMID:26261057

  13. Cloning of TRP Gene of Bombyx mori%家蚕TPR基因的克隆

    Institute of Scientific and Technical Information of China (English)

    牛伟涛

    2011-01-01

    [ Objective ] The aim was to clone Bornbyx rnori TRP gene. [ Method ] The total RNA of Bombyx mori was extracted during its pupal pedod with Trizol method, and then the TRP gene was cloned by RT-PCR. [ Result] The full-length cDNA of TRP gene was successfully obtained, and the ORF fragment (858 bp) of TRP gene was cloned by PCR. [ Conclusion] TRP gene of Bombyx mori was cloned for the first time,which could provide solid basis for the study on its function.%[目的]克隆家蚕TPR基因,为研究该基因功能提供依据.[方法]利用Trizol法提取家蚕蛹期总RNA,再用RT-PCR的方法对家蚕TPR基因进行克隆.[结果]成功得到家蚕TPR基因全cDNA序列,并用PCR扩增得到了TPR基因的ORF,全长858 bp.[结论]首次对家蚕TPR家族基因进行了克隆,为今后对该基因功能研究奠定了坚实的基础.

  14. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  16. 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.

  17. 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.

  18. 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.

  19. 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

  20. 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

  1. 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.  

  2. 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.

  3. Microstructural, thermal and antibacterial properties of electron beam irradiated Bombyx mori silk fibroin films

    Energy Technology Data Exchange (ETDEWEB)

    Asha, S.; Sanjeev, Ganesh, E-mail: ganeshsanjeev@rediffmail.com [Microtron Center, Department of Studies in Physics, Mangalore University, Mangalagangotri - 574199 (India); Sangappa [Department of Studies in Physics, Mangalore University, Mangalagangotri - 574199 (India); Naik, Prashantha; Chandra, K. Sharat [Department of Biosciences, Mangalore University, Mangalagangotri - 574199 (India)

    2014-04-24

    The Bombyx mori silk fibroin (SF) films were prepared by solution casting method and the effects of electron beam on structural, thermal and antibacterial responses of the prepared films were studied. The electron irradiation for different doses was carried out using 8 MeV Microtron facility at Mangalore University. The changes in microstructural parameters and thermal stability of the films were investigated using Wide Angle X-ray Scattering (WAXS) and thermogravimetric analysis (TGA) respectively. Both microstructuralline parameters (crystallite size and lattice strain (g in %)) and thermal stability of the irradiated films have increased with radiation dosage. Agar diffusion method demonstrated the antibacterial activity of SF film which was increased after irradiation on both Gram-positive and Gram-negative species.

  4. 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

  5. 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

  6. Transgene-based, female-specific lethality system for genetic sexing of the silkworm, Bombyx mori.

    Science.gov (United States)

    Tan, Anjiang; Fu, Guoliang; Jin, Li; Guo, Qiuhong; Li, Zhiqian; Niu, Baolong; Meng, Zhiqi; Morrison, Neil I; Alphey, Luke; Huang, Yongping

    2013-04-23

    Transgene-based genetic sexing methods are being developed for insects of agricultural and public health importance. Male-only rearing has long been sought in sericulture because males show superior economic characteristics, such as better fitness, lower food consumption, and higher silk yield. Here we report the establishment of a transgene-based genetic sexing system for the silkworm, Bombyx mori. We developed a construct in which a positive feedback loop regulated by sex-specific alternative splicing leads to high-level expression of the tetracycline-repressible transactivator in females only. Transgenic animals show female-specific lethality during embryonic and early larval stages, leading to male-only cocoons. This transgene-based female-specific lethal system not only has wide application in sericulture, but also has great potential in lepidopteran pest control. PMID:23569267

  7. 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.

  8. 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

  9. 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.

  10. 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

  11. 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.

  12. 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.

  13. Studies on the Mutant Systems of the Bombyx mori Gene Bank

    Institute of Scientific and Technical Information of China (English)

    LU Cheng; DAI Fang-yin; XIANG Zhong-huai

    2002-01-01

    Through over ten years of study, more than 1 000 genetic materials including mutant genes,chromosomal variation strains and special genetic materials of Bombyx mori, Linnaeus, collected, introduced or created since 1940s especially late 1980s, have been sorted out and put in order. After identifications and genetic analyses of their morphological, physiological and biochemical characters, the silkworm gene bank was constructed and the preservation system was perfected, and more than 600 silkworm strains were kept in this gene bank. The preserved silkworm mutant genes have covered more than 90% of existent ones across the world, in which, more than 100 are rare and precious mutant genes, and over 60 mutant genes were found and studied for the first time. Through hybrid analyses, linkage tests and three-point gene location tests, a perfect linkage retrieval labeling system of silkworm was established, which included 230 marker genes covering all the 28 linkage groups of Bombyx mori. The gene location system (composite system of recessive genes) of different linkage groups was set up. The intergenic complementation of mutant egg color and third type of maternal heredity egg color have been found, and indicated that the epistatic effect of mutant gene of white egg is universal. Twenty eight independent near isogenic lines murked with morphological mutation gene have been created and a series of novel breeding materials possessing great potential for application such as high feeding efficiency, special sex markers, natural colored silk, resistance to disease, wider feeding range and adjustable parthenogenesis, etc., have been developed. The sustainable maintenance and management technique system of silkworm gene resources were well established.

  14. Molecular cloning of pheromone biosynthesis activating neuropeptide in silkworm, Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    徐卫华; Yukihiro Stao; Okitsugu Yamashita

    1996-01-01

    Pheromone biosynthesis activating neuropeptide (PBAN) is a suboesophageal ganglion secretory polypeptide of insect, which activates the pheromone gland to produce sex pheromone biosynthesis in female silkworm, Bombyx mori. A Bombyx genomic library was screened by the method of plaque hybridization using the 32P-labeled BomDH cDNA as a probe. The genomic sequence encoding PBAN has been cloned and its structure is analyzed. The PBAN gene comprises two exons interspersed by a single intron 697 bp in length. Preceding the PBAN amino acid sequence is a 32-amino acid sequence containing two FXPRL amide peptides, which are α-SGNP (Ile-Ile-Phe-Thr-Pro-Lys-Leu) and β-SGNP (Ser-Val-Ala-Asn-Pro-Arg-Thr-His-Glu-Ser-Leu-Glu-Phe-Ile-Pro-Arg-Leu), which is followed by a Gly-Arg processing site. Immediately, after the PBAN amino acid sequence is a Gly-Arg processing site and a FXPRL amide peptide γ-SGNP (Thr-Met-Ser-Phe-Ser-Pro-Arg-Leu). It is suggested that besides PBAN, 7-, 8-, and 17-residue amidated peptides wer

  15. 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.

  16. Identification and Molecular Characterization of a Chitin Deacetylase from Bombyx mori Peritrophic Membrane

    Directory of Open Access Journals (Sweden)

    Xiao-Wu Zhong

    2014-01-01

    Full Text Available The insect midgut epithelium is generally lined with a unique chitin and protein structure, the peritrophic membrane (PM, which facilitates food digestion and protects the gut epithelium. PM proteins are important determinants for PM structure and formation. In this study, the silkworm Bombyx mori midgut PM protein BmCDA7 was identified by proteomic tools. The full-length BmCDA7 cDNA is 1357 bp; the deduced protein is composed of 379 amino acid residues and includes a 16 amino acid residue signal peptide, a putative polysaccharide deacetylase-like domain and 15 cysteine residues present in three clusters. The heterologously expressed proteins of the BmCDA7 gene in yeast displayed chitin deacetylase activity. Expression of B. mori BmCDA7 was detected in the midgut at both the transcriptional and translational levels. The BmCDA7 gene was expressed by the newly hatched silkworm larvae until day seven of the fifth instar and was expressed at a high level in the newly exuviated larvae of different instars. The functions and regulatory mechanism of BmCDA7, however, need further investigation.

  17. 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.

  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. 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...

  20. 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.

  1. 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.

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

    Directory of Open Access Journals (Sweden)

    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

  3. A hypothetical model of crossing Bombyx mori nucleopolyhedrovirus through its host midgut physical barrier.

    Directory of Open Access Journals (Sweden)

    Yang Cheng

    Full Text Available Bombyx mori nucleopolyhedrovirus (BmNPV is a primary pathogen of silkworm (B. mori that causes severe economic losses each year. However, the molecular mechanisms of silkworm-BmNPV interactions, especially the silkworm proteins that can interact with the virus, are still largely unknown. In this study, the total and membrane proteins of silkworm midguts were displayed using one- and two-dimensional electrophoresis. A virus overlay assay was used to detect B. mori proteins that specifically bind to BmNPV particles. Twelve proteins were located and identified using mass spectrometry, and the different expression of the corresponding genes in BmNPV susceptible and resistant silkworm strains also indicated their involvement in BmNPV infection. The 12 proteins are grouped based on their potential roles in viral infection, for example, endocytosis, intracellular transportation, and host responses. Based on these results, we hypothesize the following: I vacuolar ATP synthase catalytic subunit A and subunit B may be implicated in the process of the membrane fusion of virus and the release of the nucleocapsid into cytoplasm; II actin, enolase and phosphoglycerate kinase are cytoskeleton associated proteins and may play an important role in BmNPV intracellular transportation; III mitochondrial prohibitin complex protein 2, ganglioside-induced differentiation-associated protein, calreticulin, regucalcin-like isoform X1 and 60 kDa heat shock protein are involved in cell apoptosis regulation during BmNPV infection in larvae midguts; IV ribosomal P0 may be associated with BmNPV infection by regulating gene expression of BmNPV; V arginine kinase has a role in the antiviral activities against BmNPV. Our work should prove informative by providing multiple protein targets and a novel direction to investigate the molecular mechanisms of the interactions between silkworms and BmNPV.

  4. Transgenic characterization of two testis-specific promoters in the silkworm, Bombyx mori.

    Science.gov (United States)

    Xu, J; Bi, H; Chen, R; Aslam, A F M; Li, Z; Ling, L; Zeng, B; Huang, Y; Tan, A

    2015-04-01

    Sex-specific regulatory elements are key components for developing insect genetic sexing systems. The current insect genetic sexing system mainly uses a female-specific modification system whereas little success was reported on male-specific genetic modification. In the silkworm Bombyx mori, a lepidopteran model insect with economic importance, a transgene-based, female-specific lethality system has been established based on sex-specific alternative splicing factors and a female-specific promoter BmVgp (vitellogenin promoter) has been identified. However, no male-specific regulatory elements have yet been identified. Here we report the transgenic identification of two promoters that drive reporter gene expression in a testis-specific manner in B. mori. Putative promoter sequences from the B. mori Radial spoke head 1 gene (BmR1) and beta-tubulin 4 gene (Bmβ4) were introduced using piggybac-based germline transformation. In transgenic silkworms, expression of the reporter gene enhanced green fluorescent protein (EGFP) directed by either BmR1 promoter (BmR1p) or Bmβ4p showed precisely testis-specific manners from the larval to adult stage. Furthermore, EGFP expression of these two transgenic lines showed different localization in the testis, indicating that BmR1p or Bmβ4p might be used as distinct regulatory elements in directing testis-specific gene expression. Identification of these testis-specific promoters not only contributes to a better understanding of testis-specific gene function in insects, but also has potential applications in sterile insect techniques for pest management. PMID:25387604

  5. 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.

  6. 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.

  7. 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)

    Bharathi,D.; 缪云根

    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.

  8. 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...

  9. 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...

  10. 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...

  11. 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...

  12. 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...

  13. 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.

  14. 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

  15. 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.

  16. 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.

  17. 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

  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. Preparation and characterization of regenerated Bombyx mori silk fibroin fiber with high strength

    Directory of Open Access Journals (Sweden)

    2008-12-01

    Full Text Available Regenerated Bombyx mori silk fibers were spun from hexafluoro-iso- propanol solution of silk fibroin sponge in methanol used as a coagulant solvent and then elongated in water. The stress-strain curves of the regenerated fibers changed dramatically depending on the draw ratio and the structure was studied by 13C CP/MAS NMR and X-ray diffraction methods. The patterns of 13C CP/MAS NMR spectra of two regenerated fibers with different draw ratios (1× and 3× and native silk fiber are all β-sheet structure although the fraction of random coil/distorted β-turn decreases in the order of 1×, 3× and native fiber gradually. On the other hand, azimuthal scans of their X-ray fiber patterns changed remarkably with increasing the draw ratio. This indicates that long-range orientation of the fibroin chain changes remarkably during the drawing process, but the short-range local structure does not change significantly. Regenerated silk fiber with a draw ratio of 3× is a fiber with high strength which is comparable with that of natural silk fiber. The regenerated fiber is also more degradable than natural silk fiber in enzyme solution in vitro.

  1. 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.

  2. Functional Loss of Bmsei Causes Thermosensitive Epilepsy in Contractile Mutant Silkworm, Bombyx mori

    Science.gov (United States)

    Nie, Hongyi; Cheng, Tingcai; Huang, Xiaofeng; Zhou, Mengting; Zhang, Yinxia; Dai, Fangyin; Mita, Kazuei; Xia, Qingyou; Liu, Chun

    2015-07-01

    The thermoprotective mechanisms of insects remain largely unknown. We reported the Bombyx mori contractile (cot) behavioral mutant with thermo-sensitive seizures phenotype. At elevated temperatures, the cot mutant exhibit seizures associated with strong contractions, rolling, vomiting, and a temporary lack of movement. We narrowed a region containing cot to ~268 kb by positional cloning and identified the mutant gene as Bmsei which encoded a potassium channel protein. Bmsei was present in both the cell membrane and cytoplasm in wild-type ganglia but faint in cot. Furthermore, Bmsei was markedly decreased upon high temperature treatment in cot mutant. With the RNAi method and injecting potassium channel blockers, the wild type silkworm was induced the cot phenotype. These results demonstrated that Bmsei was responsible for the cot mutant phenotype and played an important role in thermoprotection in silkworm. Meanwhile, comparative proteomic approach was used to investigate the proteomic differences. The results showed that the protein of Hsp-1 and Tn1 were significantly decreased and increased on protein level in cot mutant after thermo-stimulus, respectively. Our data provide insights into the mechanism of thermoprotection in insect. As cot phenotype closely resembles human epilepsy, cot might be a potential model for the mechanism of epilepsy in future.

  3. 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

  4. 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

  5. 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

  6. 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.

  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. 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

  9. 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

  10. 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.

  11. 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.

  12. 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.

  13. 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.

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

    Science.gov (United States)

    George, Karina A; Shadforth, Audra M A; Chirila, Traian V; Laurent, Matthieu J; Stephenson, Sally-Anne; Edwards, Grant A; Madden, Peter W; Hutmacher, Dietmar W; Harkin, Damien G

    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.

  15. 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

  16. Sorbitol as an arrester of embryonic development in diapausing eggs of the silkworm, Bombyx mori.

    Science.gov (United States)

    Horie; Kanda; Mochida

    2000-06-01

    Recently, it was confirmed that embryos derived from diapausing eggs of the silkworm, Bombyx mori, begin their development and reach larval maturity on mulberry leaves, when the naked eggs are cultured in vitro. In this study, we found that the method of embryo culture is useful for determining the physiological regulation of diapause. We show that the development of embryos derived from diapausing eggs was strongly inhibited by the addition of either sorbitol or trehalose to the culture medium. Furthermore, this inhibitory effect disappeared when the embryos were cultured in a control medium which did not contain either sorbitol or trehalose, indicating that the inhibitory reactions caused by both substances are reversible. The minimal effective dose of either sorbitol or trehalose was approximately 0.2 M, a value similar to the in vivo concentration of sorbitol in diapausing eggs (0.2 M). Glycerol, mannitol or glucose were moderately effective for inhibition. Sorbitol present in diapausing silkworm eggs does not appear to serve as an antifreeze, but as an strong arresting factor of embryonic development. Furthermore, these results show that a decrease in sorbitol releases the embryos from diapause at the termination of diapause.

  17. 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.

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

    Energy Technology Data Exchange (ETDEWEB)

    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.

  19. Identification of a Serratia marcescens virulence factor that promotes hemolymph bleeding in the silkworm, Bombyx mori.

    Science.gov (United States)

    Ishii, Kenichi; Adachi, Tatsuo; Hara, Takashi; Hamamoto, Hiroshi; Sekimizu, Kazuhisa

    2014-03-01

    Injection of culture supernatant of Serratia marcescens, a Gram-negative bacterium pathogenic to a wide range of host animals including insects and mammals, into the hemolymph of silkworm (Bombyx mori) larvae led to continuous flow of the hemolymph (blood of insects) from the injection site. The amount of hemolymph lost within 60 min reached 15-20% of the total larval weight. Using a bioassay with live silkworms, we purified Serralysin, a metalloprotease that requires divalent cations for its activity, as the factor responsible for the promotion of hemolymph bleeding from the culture supernatant of S. marcescens. Recombinant protein also induced hemolymph bleeding in silkworms. Moreover, the culture supernatant of an S. marcescens disruption mutant of the ser gene showed attenuated ability to promote hemolymph bleeding. In addition, this bleeding-promoting activity of the S. marcescens culture supernatant was attenuated by disruption of the wecA gene, which is involved in the biosynthesis of the lipopolysaccharide O-antigen. These findings suggest that Serralysin metalloprotease contributes to the pathogenesis of S. marcescens by inhibiting wound healing, which leads to a massive loss of hemolymph from silkworm larvae.

  20. 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.

  1. 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

  2. A novel sucrose hydrolase from the bombycoid silkworms Bombyx mori, Trilocha varians, and Samia cynthia ricini with a substrate specificity for sucrose.

    Science.gov (United States)

    Wang, Huabing; Kiuchi, Takashi; Katsuma, Susumu; Shimada, Toru

    2015-06-01

    Although membrane-associated sucrase activity has been detected in the midgut of various lepidopteran species, it has not yet been identified and characterized at the molecular level. In the present study, we identified a novel sucrose hydrolase (SUH) gene from the following three bombycoid silkworms: Bombyx mori, Trilocha varians, and Samia cynthia ricini and named them BmSuh, TvSuh, and ScSuh, respectively. The EST dataset showed that BmSuh is one of the major glycoside hydrolase genes in the larval midgut of B. mori. These genes were almost exclusively expressed in the larval midgut in all three species, mainly at the feeding stage. SUHs are classified into the glycoside hydrolase family 13 and show significant homology to insect maltases. Enzymatic assays revealed that recombinant SUHs were distinct from conventional maltases and exhibited substrate specificity for sucrose. The recombinant BmSUH was less sensitive to sugar-mimic alkaloids than TvSUH and ScSUH, which may explain the reason why the sucrase activity in the B. mori midgut was less affected by the sugar-mimic alkaloids derived from mulberry.

  3. 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 < 100nm). SWNT-silk multifunctional nanocomposite fibers were fabricated for the first time with anticipated properties (mechanical, thermal and electrically conductive) that may have scientific applications (nerve regeneration, stimulation of cell-scaffold interaction). In order to realize these applications, the following areas need to be addressed: a systematic investigation of the dispersion of the nanotubes in the silk matrix, a determination of new methodologies for characterizing the nanofiber properties and establishing the

  4. 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...

  5. 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...

  6. 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

  7. 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

  8. 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.

  9. 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.

  10. Transcriptional profiling of midgut immunity response and degeneration in the wandering silkworm, Bombyx mori.

    Directory of Open Access Journals (Sweden)

    Qiuyun Xu

    Full Text Available BACKGROUND: Lepidoptera insects have a novel development process comprising several metamorphic stages during their life cycle compared with vertebrate animals. Unlike most Lepidoptera insects that live on nectar during the adult stage, the Bombyx mori silkworm adults do not eat anything and die after egg-laying. In addition, the midguts of Lepidoptera insects produce antimicrobial proteins during the wandering stage when the larval tissues undergo numerous changes. The exact mechanisms responsible for these phenomena remain unclear. PRINCIPAL FINDINGS: We used the silkworm as a model and performed genome-wide transcriptional profiling of the midgut between the feeding stage and the wandering stage. Many genes concerned with metabolism, digestion, and ion and small molecule transportation were down-regulated during the wandering stage, indicating that the wandering stage midgut loses its normal functions. Microarray profiling, qRT-PCR and western blot proved the production of antimicrobial proteins (peptides in the midgut during the wandering stage. Different genes of the immune deficiency (Imd pathway were up-regulated during the wandering stage. However, some key genes belonging to the Toll pathway showed no change in their transcription levels. Unlike butterfly (Pachliopta aristolochiae, the midgut of silkworm moth has a layer of cells, indicating that the development of midgut since the wandering stage is not usual. Cell division in the midgut was observed only for a short time during the wandering stage. However, there was extensive cell apoptosis before pupation. The imbalance of cell division and apoptosis probably drives the continuous degeneration of the midgut in the silkworm since the wandering stage. CONCLUSIONS: This study provided an insight into the mechanism of the degeneration of the silkworm midgut and the production of innate immunity-related proteins during the wandering stage. The imbalance of cell division and apoptosis

  11. 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...

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    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. 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.

  15. 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

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

    Directory of Open Access Journals (Sweden)

    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.

  17. 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

  18. 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

  19. 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

  20. 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.

  1. 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

  2. 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.

  3. Shotgun proteomic analysis of the Bombyx mori anterior silk gland: An insight into the biosynthetic fiber spinning process.

    Science.gov (United States)

    Yi, Qiying; Zhao, Ping; Wang, Xin; Zou, Yong; Zhong, Xiaowu; Wang, Chen; Xiang, Zhonghuai; Xia, Qing-You

    2013-09-01

    The Bombyx mori anterior silk gland (ASG) is a natural fiber manipulator for the material provided by the middle and posterior silk glands. In view of the significant role of the ASG in the liquid-crystal spinning process, a shotgun proteomics approach was taken to study the relationship between the function of proteins in the silkworm ASG and the spinning mechanism. A total of 1132 proteins with 7647 unique peptides were identified in the ASG dataset including some involved in the cuticle, ion transportation, energy metabolism, and apoptosis. Two putative cuticle-specific proteins were highly and specifically expressed in the ASG; therefore, the ASG dataset could provide clues for comprehensive understanding of the natural silk spinning mechanism in the silkworm. All MS data have been deposited in the ProteomeXchange with identifier PXD000090.

  4. 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.

  5. 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

  6. 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

  7. 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...

  8. 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.

  9. 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

  10. 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...

  11. Comparative Study of Biological Characteristics of Larvae, Crude and Dried Cocoon in 7 Races of Silkworm Bombyx mori L., Raised in Transylvania Area

    Directory of Open Access Journals (Sweden)

    Emilia Maria Furdui

    2010-05-01

    Full Text Available 7 monovoltine races of Romanian silkworm Bombyx mori L. were breeded in specific Transylvania conditions, assuring the same microclimate conditions, being fed with the same mulberry leaves (Ukraina 107. The results obtained showed a high homogeneity, the differences were due to the variability and genotype characteristic for each individual of every race. Results and standard deviations are within the race standard. The study permitted a ierarchy of the races, following the biological characteristics, microclimate conditions, quantity, quality and breeding technology.

  12. 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

  13. 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

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

    Directory of Open Access Journals (Sweden)

    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.

  15. 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

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

    Directory of Open Access Journals (Sweden)

    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.

  17. 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

  18. 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

  19. Genome-Wide Identification, Characterization and Expression Analysis of the Solute Carrier 6 Gene Family in Silkworm (Bombyx mori

    Directory of Open Access Journals (Sweden)

    Xin Tang

    2016-10-01

    Full Text Available The solute carrier 6 (SLC6 gene family, initially known as the neurotransmitter transporters, plays vital roles in the regulation of neurotransmitter signaling, nutrient absorption and motor behavior. In this study, a total of 16 candidate genes were identified as SLC6 family gene homologs in the silkworm (Bombyx mori genome. Spatio-temporal expression patterns of silkworm SLC6 gene transcripts indicated that these genes were highly and specifically expressed in midgut, brain and gonads; moreover, these genes were expressed primarily at the feeding stage or adult stage. Levels of expression for most midgut-specific and midgut-enriched gene transcripts were down-regulated after starvation but up-regulated after re-feeding. In addition, we observed that expression levels of these genes except for BmSLC6-15 and BmGT1 were markedly up-regulated by a juvenile hormone analog. Moreover, brain-enriched genes showed differential expression patterns during wandering and mating processes, suggesting that these genes may be involved in modulating wandering and mating behaviors. Our results improve our understanding of the expression patterns and potential physiological functions of the SLC6 gene family, and provide valuable information for the comprehensive functional analysis of the SLC6 gene family.

  20. 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.

  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. 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.

  3. Genome-Wide Identification, Characterization and Expression Analysis of the Solute Carrier 6 Gene Family in Silkworm (Bombyx mori)

    Science.gov (United States)

    Tang, Xin; Liu, Huawei; Chen, Quanmei; Wang, Xin; Xiong, Ying; Zhao, Ping

    2016-01-01

    The solute carrier 6 (SLC6) gene family, initially known as the neurotransmitter transporters, plays vital roles in the regulation of neurotransmitter signaling, nutrient absorption and motor behavior. In this study, a total of 16 candidate genes were identified as SLC6 family gene homologs in the silkworm (Bombyx mori) genome. Spatio-temporal expression patterns of silkworm SLC6 gene transcripts indicated that these genes were highly and specifically expressed in midgut, brain and gonads; moreover, these genes were expressed primarily at the feeding stage or adult stage. Levels of expression for most midgut-specific and midgut-enriched gene transcripts were down-regulated after starvation but up-regulated after re-feeding. In addition, we observed that expression levels of these genes except for BmSLC6-15 and BmGT1 were markedly up-regulated by a juvenile hormone analog. Moreover, brain-enriched genes showed differential expression patterns during wandering and mating processes, suggesting that these genes may be involved in modulating wandering and mating behaviors. Our results improve our understanding of the expression patterns and potential physiological functions of the SLC6 gene family, and provide valuable information for the comprehensive functional analysis of the SLC6 gene family. PMID:27706106

  4. 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

  5. Production Efficiency of Cocoon Shell of Silkworm, Bombyx mori L. (Bombycidae: Lepidoptera, as an Index for Evaluating the Nutritive Value of Mulberry, Morus sp. (Moraceae, Varieties

    Directory of Open Access Journals (Sweden)

    Jalaja Suresh Kumar

    2011-01-01

    Full Text Available The nutritional efficiency of mulberry leaves consumed by silkworms, Bombyx mori L., is usually evaluated in terms of the proportion of cocoon shell weight to the amount of food ingested. The production efficiency of cocoon shell is generally used to identify the superiority of a mulberry variety for silkworm rearing. In this study the production efficiency of cocoon shell was used as an index for evaluating the nutritive value of different mulberry varieties of India. Among the varieties, V-1, having highest production efficiency of cocoon shell with less amount of food ingested and highest digestibility, is regarded as the best suitable variety with nutritive values ideal for silkworm rearing.

  6. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

  9. 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

  10. 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

  11. 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.

  12. 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.

  13. 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

  14. 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...

  15. Glutathione-binding site of a bombyx mori theta-class glutathione transferase.

    Science.gov (United States)

    Hossain, M D Tofazzal; Yamada, Naotaka; Yamamoto, Kohji

    2014-01-01

    The glutathione transferase (GST) superfamily plays key roles in the detoxification of various xenobiotics. Here, we report the isolation and characterization of a silkworm protein belonging to a previously reported theta-class GST family. The enzyme (bmGSTT) catalyzes the reaction of glutathione with 1-chloro-2,4-dinitrobenzene, 1,2-epoxy-3-(4-nitrophenoxy)-propane, and 4-nitrophenethyl bromide. Mutagenesis of highly conserved residues in the catalytic site revealed that Glu66 and Ser67 are important for enzymatic function. These results provide insights into the catalysis of glutathione conjugation in silkworm by bmGSTT and into the metabolism of exogenous chemical agents.

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

    Directory of Open Access Journals (Sweden)

    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.

  17. Metabolomics Analysis of the Larval Head of the Silkworm, Bombyx mori

    Science.gov (United States)

    Li, Yi; Wang, Xin; Chen, Quanmei; Hou, Yong; Xia, Qingyou; Zhao, Ping

    2016-01-01

    The head, which performs many biological functions, is the most complicated structure of an insect. Development, locomotor behavior, food intake, environmental sensing, and signal transduction are all controlled by the insect’s head. As a well-studied insect in Lepidoptera, the silkworm head has an additional function of spinning silk fibers. To understand which molecules are involved in these physiological activities, we performed a metabolomics analysis of silkworm heads. By integrating GC-MS and LC-MS/MS, 90 metabolites were identified in the larval heads of silkworms. These were classified into 13 categories, including amino acids, sugars, organic acids, nucleotides, alcohols, and fatty acids. Informatics analysis revealed that these metabolites are involved in cellular processes, environmental information processing, genetic information processing, human diseases, metabolism, organismal systems, and other pathways. The identified metabolites and pathways are involved in biological processes such as signal transduction, carbohydrate metabolism, endocrine activities, and sensory activities; reflecting the functions of various organs in silkworm heads. Thus, our findings provide references which elucidate the potential functions of the silkworm head and will be of great value for the metabolomics research of silkworms and other insects. PMID:27657048

  18. 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

  19. Transcriptional analysis of sex pheromone biosynthesis signal genes in Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    Shi-Heng An; Meng-Fang Du; Li-Juan Su; Xin-Ming Yin

    2012-01-01

    Six sex pheromone synthesis signal genes,including acyl coenzyme A (acylCoA) desaturase (desatl),fatty acyl reductase (FAR),pheromone biosynthesis activating neuropeptide receptor (PBANR),fatty acid transport protein (FATP),acyl-CoA binding protein (ACBP) and store-operated channel protein (OrailA),were studied for their transcriptional regulations.The expression profiles of these transcripts at different developmental stages (from-96 to 48 h) revealed that the genes are expressed in an age-dependent manner.The transcripts of these genes continued to increase despite decapitation,and compared with normally developmental females,decapitation significantly inhibited their expression.Further experiments with a methoprene,a juvenile hormone (JH) analogue,challenge showed that JH was not a key inhibiting factor in the expression of these genes,and mating was found to significantly inhibit the expression of these marker genes.Altogether,the results provide a reference for understanding the mechanism of sex pheromone synthesis.

  20. The Odorant Binding Protein Gene Family from the Genome of Silkworm, Bombyx mori

    Directory of Open Access Journals (Sweden)

    Zhao Ping

    2009-07-01

    Full Text Available Abstract Background Chemosensory systems play key roles in the survival and reproductive success of insects. Insect chemoreception is mediated by two large and diverse gene superfamilies, chemoreceptors and odorant binding proteins (OBPs. OBPs are believed to transport hydrophobic odorants from the environment to the olfactory receptors. Results We identified a family of OBP-like genes in the silkworm genome and characterized their expression using oligonucleotide microarrays. A total of forty-four OBP genes were annotated, a number comparable to the 57 OBPs known from Anopheles gambiae and 51 from Drosophila melanogaster. As seen in other fully sequenced insect genomes, most silkworm OBP genes are present in large clusters. We defined six subfamilies of OBPs, each of which shows lineage-specific expansion and diversification. EST data and OBP expression profiles from multiple larvae tissues of day three fifth instars demonstrated that many OBPs are expressed in chemosensory-specific tissues although some OBPs are expressed ubiquitously and others exclusively in non-chemosensory tissues. Some atypical OBPs are expressed throughout development. These results reveal that, although many OBPs are chemosensory-specific, others may have more general physiological roles. Conclusion Silkworms possess a number of OBPs genes similar to other insects. Their expression profiles suggest that many OBPs may be involved in olfaction and gustation as well as general carriers of hydrophobic molecules. The expansion of OBP gene subfamilies and sequence divergence indicate that the silkworm OBP family acquired functional diversity concurrently with functional constraints. Further investigation of the OBPs of the silkworm could give insights in the roles of OBPs in chemoreception.

  1. 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

    Directory of Open Access Journals (Sweden)

    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%.

  2. 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

  3. 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

  4. 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)

  5. 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.

  6. 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

  7. Research advances in cytochrome P450 genes in the silkworm, Bombyx mori%家蚕细胞色素P450基因的研究进展

    Institute of Scientific and Technical Information of China (English)

    艾均文; 薛宏; 何行健; 孟繁利; 朱勇; 向仲怀

    2011-01-01

    The cytochrome P450 monooxygenases ( P450s, C Yps) constitute a large and complex superfamily of heme-thiolate proteins, which are responsible for the oxidative metabolism of structurally diverse endogenous and exogenous compounds. In this review, recent progress in Bombyx mori P450 diversity, multiple functions, genomic distribution, intron-exon organization and the evolutionary relationships to P450s from Drosophila melanogaster is summarized, and the proposals of B. Mori P450 study are also put forward. In the silkworm, the paralog count for P450s is lower than those found in other scavengers and omnivorous phytophagous insects, but substantially higher than that observed in Apis mellifera. The distribution of B. Mori P450s in the genome indicates that most of them are tandem arranged on chromosomes. There is a relatively good correlation between intron-exon organization and phylogenetic relationship among these multiple P450s. Comparison of the P450s from B. Mori to the P450s from D. Melanogaster reveals that there are 10 pairs of recognizable orthologs and the P450s in CYP3 and CYP4 clans are present with species-specific expansion. These diverse P450s have been demonstrated to be associated with growth and development, tolerance to fluoride and resistance to insecticides. The silkworm is a good representative insect of the order Lepidoptera, and it is so expected that it might found theoretical basis and model system for developmental regulation and resistance management of other insects, especially for lepidopteran insects, with the further study of B. Mori P450s.%细胞色素P450(P450s,cYPs)超基因家族是由数量众多、功能复杂的一类血红蛋白酶基因所组成,对许多结构多样的外源与内源化合物起着氧化代谢的作用.本文系统地综述了家蚕Bombyx mori基因组中P450s的数量与种类,其数量比食腐昆虫和杂食性的植食昆虫少,但比意大利蜜蜂Apis mellifera多,在基因组中大多呈串联重复

  8. 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

  9. 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%.

  10. 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

  11. 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

  12. 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.

  13. Change in kidney damage biomarkers after 13 weeks of exposing rats to the complex of Paecilomyces sinclairii and its host Bombyx mori larvae.

    Science.gov (United States)

    Jeong, Mihye; Kim, Young-Won; Min, Jeong-Ran; Kwon, Min; Han, Beom-Suk; Kim, Jeong-Gyu; Jeong, Sang-Hee

    2013-09-01

    Complex of Paecilomyces sinclairii and host larvae, Bombyx mori, is a well known health food; however, concerns about nephrotoxicity have been raised. Kidney toxicity was investigated after 13 weeks of administering the complex orally to rats with parameters including blood urea nitrogen (BUN), creatinine, and kidney damage biomarkers, beta-2-microglobulin (β2m), glutathione S-transferase alpha (GST-α), kidney injury molecule 1 (KIM-1), tissue inhibitor of matrix metalloproteinase 1 (TIMP-1), vascular endothelial growth factor (VEGF), calbindin, clusterin, cystatin C, neutrophil gelatinase-associated lipocalin (NGAL), and osteopontin. Dose-dependent kidney cell karyomegaly and tubular hypertrophy were observed, with higher severity in males. There was a dose-dependent increase in KIM-1 and TIMP-1 levels in kidney and urinary KIM-1, cystatin C, β2m, and osteopontin levels. KIM-1 and TIMP-1 increased in male kidneys had not recovered by 2 weeks after stopping exposure. Cystatin C in kidney was significantly lowered in all treatment groups at 13 weeks of administration. All the changes were more noticeable in males. These data indicate that the complex damage renal tubule cells with histopathological lesions and changes in biomarker levels. Kidney and urinary KIM-1 and cystatin C were the most markedly affected and early increased indicators among biomarkers tested, whereas BUN and creatinine were not affected.

  14. 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

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

    Directory of Open Access Journals (Sweden)

    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.

  16. 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

  17. 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)

  18. 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,

  19. 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...

  20. 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...

  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. Bombyx mori nucleopolyhedrovirus nucleic acid binding proteins BRO-B and BRO-E associate with host T-cell intracellular antigen 1 homologue BmTRN-1 to influence protein synthesis during infection.

    Science.gov (United States)

    Kotani, Eiji; Muto, Sayaka; Ijiri, Hiroshi; Mori, Hajime

    2015-07-01

    Previous reports have indicated that the Bombyx mori nucleopolyhedrovirus (BmNPV) nucleic acid binding proteins BRO-B and BRO-E are expressed during the early stage of infection and that the BRO family likely supports the regulation of mRNA; however, no study has directly examined the function of BRO family proteins in virus-permissive cells. Here, we show that BRO-B and BRO-E associate with cellular T-cell intracellular antigen 1 homologue (BmTRN-1), a translational regulator, and other cellular translation-related proteins in silkworm cells during viral infection. We created BM-N cells that expressed BRO-B/E to study molecular interactions between BmTRN-1 and BRO-B/E and how they influenced protein synthesis. Fluorescent microscopy revealed that BmTRN-1 was localized in cytoplasmic foci during BmNPV infection. Immunofluorescence studies confirmed that BmTRN-1 and BRO-B/E were colocalized in the amorphous conspicuous cytoplasmic foci. Reporter gene studies revealed that co-expression of BRO-B/E synergistically led to a significant decrease in protein synthesis from a designed transcript carrying the 5'untranslated region of a cellular mRNA with no significant change of transcript abundance. Additionally, RNA interference-mediated knockdown of BmTRN-1 resulted in a marked inhibition of the ability of BRO-B/E to regulate the transcript. These results suggested that the association of BmTRN-1 with BRO-B/E is responsible for the inhibitory regulation of certain mRNAs at the post-transcriptional level and add an additional mechanism for how baculoviruses control protein synthesis during infection.

  3. Identification and comparative analysis of immune-related genes and signaling pathways in the silkworm, Bombyx mori%家蚕免疫相关基因和信号途径的鉴定和比较分析

    Institute of Scientific and Technical Information of China (English)

    程廷才; 夏庆友; 许平震; 谭祥; 方婷; 向仲怀

    2009-01-01

    The silkworm,Bombyx mori,has been a domesticated,economically important insect for 5 000 years.Recent accomplishments in molecular immunology have revealed just a preliminary outline for silkworm innate immunity.The acquisition of the updated silkworm genome has enabled a comparative analysis of the silkworm immune-related genes and signaling pathways.In this study,through comparing with the sequenced Drosophila melanogaster,Anopheles gambiae,Apis mellifera and Tribolium castaneum genomes,we identified over 218 genes in the silkworm that fall into 21 families involved in immune defense,including pattern recognition receptors,signaling transducers,effectors and oxidative defense enzymes.Phylogenetic analysis showed that the signal transducers have remarkable orthologous relationships between different insect species in spite of the divergent sequences.In contrast,gene families associated with recognition,modulation and effectors exhibit more significant sequence conservation.However,the orthologs of these families are remarkably absent,presumably attributable to the lineage-specific gene duplication.Our results suggest that common mechanisms may be responsible for innate-immunity responses to pathogens via signaling pathways in the silkworm.Furthermore,hosts may adjust their defensive strategies by gene duplication and sequence divergence to kill pathogens.%家蚕 Bombyx mori 是一种重要的经济昆虫,在中国约有5 000年的驯化历史.家蚕分子免疫学方面的最新研究已经初步勾勒出其先天免疫的轮廓.本研究基于更新的家蚕基因组数据,通过与黑腹果蝇 Drosophila melanogaster、冈比亚按蚊 Anopheles gambiae、意大利蜜蜂 Apis mellifera 和赤拟谷盗 Tribolium castaneum 基因组的比较分析,鉴定了家蚕21个免疫相关基因家族的218个基因,其编码产物包括模式识别受体、信号传导因子、效应分子和氧化防御相关的酶类.尽管信号传导因子的序列

  4. 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.

  5. 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.

  6. 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.

  7. 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

  8. 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

  9. An Analysis on clip Serine Protease Family Genes Involved in Innate Immunity of Bombyx mori%参与家蚕免疫反应的clip丝氨酸蛋白酶家族基因分析

    Institute of Scientific and Technical Information of China (English)

    陈建平; 赵萍; 董照明; 张艳; 石虎; 夏庆友

    2012-01-01

    丝氨酸蛋白酶在昆虫的新陈代谢和生长发育等多种生理过程中起重要作用,特别是含clip结构域的丝氨酸蛋白酶与昆虫的免疫级联激活途径密切相关.为探索家蚕clip丝氨酸蛋白酶在家蚕先天免疫反应信号通路中的作用,对家蚕(Bombyx mori)与烟草天蛾(Manduca sexta)中鉴定获得的含有clip结构域的丝氨酸蛋白酶进行序列比对和系统进化分析,发现BmSP78与MsPAP-1、BmSP127与MsHP8、BmSP124与MsHP1、BmSP125与MsHP17位于同一进化支上,4对clip丝氨酸蛋白酶的氨基酸序列相似度分别为70%、68%、81%和57%,另外的8个蛋白酶BmSP23、BmSP111、BmSP95、BmSP135、BmSP129、Bm-SP137、BmSP91和BmSP99构成了家蚕特有的一群clip丝氨酸蛋白酶.用革兰阴性细菌沙雷氏菌和革兰阳性细菌黑胸败血菌分别注射感染家蚕5龄第4天幼虫后,通过半定量RT-PCR检测分析clip丝氨酸蛋白酶基因在不同感染时间点的表达特征:病原菌诱导后蚕体中有11个clip丝氨酸蛋白酶基因mRNA转录水平有明显变化,其中BmSP102和BmSP96于黑胸败血菌诱导12h后在血细胞中明显上调表达,而BmSP125在沙雷氏菌和黑胸败血菌感染12h后的脂肪体中均明显上调表达.研究结果为建立家蚕clip丝氨酸蛋白酶可能参与的免疫级联反应路径提供了重要线索.%Serine proteases are involved in various physiological processes of insect,such as metabolism,growth and development.Especially,serine proteases with clip domain are closely related with the activation of immune cascade pathways of insect.In order to understand the roles of serine proteases in the signal pathways of innate immune reaction in silkworm (Bombyx mori),we performed sequence alignment and phylogenetic analysis to clip serine proteases identified in B.mori and Manduca sexta.The result showed that BmSP78 and MsPAP-1,BmSP127 and MsHP8,BmSP124 and MsHP1,BmSP125 and MsHP17 were located on the same phyletic clade

  10. 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.

  11. Evolutionary Pattern of Three Bombyx mori Antimicrobial Peptide Genes Under Influence of Domestication%驯化影响下的家蚕3种抗菌肽基因的进化模式

    Institute of Scientific and Technical Information of China (English)

    郭意; 孙伟; 程静; 沈以红; 张泽; 向仲怀

    2011-01-01

    As innate immune effectors, antimicrobial peptides play crucial roles in the evolution of insect species. In this study, three different antimicrobial peptide genes,DefA,CecE and MorB3, were sequenced from both Bombyx mori and its wild relative, Bombyx mandarina. Polymorphism analyses, neutrality tests, coalescent simulation analyses and linkage disequilibrium analysis to the obtained nucleotide sequences indicated that the three genes were subject to different evolutionary patterns: DefA was a target gene of artificial selection during domestication, CecE was a neutral gene that had undergone domestication bottleneck effect, and the major driven force of MorB3 was genetic drift. Despite of these varied evolutionary patterns, the three genes had higher levels of linkage disequilibrium in domesticated silkworm than those in wild silkworm, indicating that domesticated silkworm had experienced a recent bottleneck effect, and decrease of population size had led to the reduction in gene recombinant rate. Our results provide important information for understanding the evolution and function of different antimicrobial peptides in domesticated silkworm.%抗菌肽作为昆虫的先天性免疫效应因子在昆虫物种进化过程中起着至关重要的作用.测定了家蚕(Bombyx mori)和野桑蚕(Bombyx mandarina)群体的3种不同类型的抗茵肽基因DefA、CecE和MorB3的序列,通过序列核苷酸多态性分析、中性检验、溯祖模拟分析和连锁不平衡分析,发现这3种抗茵肽基因呈现不同的进化模式:DefA属于驯化过程中人工选择的靶基因;CecE是经历了驯化瓶颈效应的中性基因;MorB3的进化受遗传漂变影响.尽管进化模式不同,但家蚕的3种抗菌肽基因连锁不平衡程度均高于野桑蚕,反映家蚕经历了瓶颈效应,群体数量减小导致基因重组率降低.这些结果为理解家蚕不同抗茵肽的进化和功能提供了重要信息.

  12. 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.

  13. 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

  14. 家蚕保幼激素甲基转移酶基因的克隆与表达分析%Cloning and Characterization of Juvenile Hormone Methyltransgerase Gene,JHAMT2,in Silkworm, Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    甘丽萍; 龙菲菲; 张进; 司马杨虎; 徐世清

    2012-01-01

    A full length cDNA, 1007 bp, was cloned according to the expression sequence tags (ESTs) of Bombyx mori. This gene contains four exons, located on the nscaf 2993 of the 12th chromosome of B. mori. The deduced protein sequence is 266aa with a functional locus of methyltransgerase super family. It owns 38 % homology with JHAMT in Manduca sexta and therefore the cloning gene was named as juvenile hormone methyltransgerase gene of Bombyx mori, JHAMT2. The JHAMT2 specially expressed in the silkgland, especially in the middle silkgland. During different developmental stages, it had a relatively higher expression level in the 4th instar larvas, while it only expressed on the third day during the whole fifth larvae. The sexual differences were reflected by a longer continuous expression time in males than that in females. The expression level of JHAMT2 was negatively regulated by exogenous MH and positively regulated by JHA during the middle stage of the 5 th instar. Males were more sensitive to exogenous insect hormones than females.%根据家蚕表达序列标签(FEST8),克隆了一个家蚕保幼激素甲基转移酶基因(JHAMT2)的全长cDNA序列.序列分析表明,该基因全长1007 bp,序列分析有4个外显子,定位于家蚕第12号染色体的nscaf2 993上,推导的编码蛋白序列长度为266AA,有一个甲基转移酶超家族功能位点,与烟草天娥(Manduca sexta)JHAMT同源性达到38%.JHAMT2基因在家蚕丝腺、特别是中部丝腺组织特异表达,发育时期表现为4龄期有比较高的表达量,5龄期只在5龄第3日有表达,性别差异表现为雄性体内持续表达时间比雌性中长.JHAMT2基因表达受到外源激素MH的负调控和JHA的正调控,雌性对外源昆虫激素的感受更敏感.

  15. Positional cloning of a Bombyx pink-eyed white egg locus reveals the major role of cardinal in ommochrome synthesis.

    Science.gov (United States)

    Osanai-Futahashi, M; Tatematsu, K-I; Futahashi, R; Narukawa, J; Takasu, Y; Kayukawa, T; Shinoda, T; Ishige, T; Yajima, S; Tamura, T; Yamamoto, K; Sezutsu, H

    2016-02-01

    Ommochromes are major insect pigments involved in coloration of compound eyes, eggs, epidermis and wings. In the silkworm Bombyx mori, adult compound eyes and eggs contain a mixture of the ommochrome pigments such as ommin and xanthommatin. Here, we identified the gene involved in ommochrome biosynthesis by positional cloning of B. mori egg and eye color mutant pink-eyed white egg (pe). The recessive homozygote of pe has bright red eyes and white or pale pink eggs instead of a normal dark coloration due to the decrease of dark ommochrome pigments. By genetic linkage analysis, we narrowed down the pe-linked region to ~258 kb, containing 17 predicted genes. RNA sequencing analyses showed that the expression of one candidate gene, the ortholog of Drosophila haem peroxidase cardinal, coincided with egg pigmentation timing, similar to other ommochrome-related genes such as Bm-scarlet and Bm-re. In two pe strains, a common missense mutation was found within a conserved motif of B. mori cardinal homolog (Bm-cardinal). RNA interference-mediated knockdown and transcription activator-like effector nuclease (TALEN)-mediated knockout of the Bm-cardinal gene produced the same phenotype as pe in terms of egg, adult eye and larval epidermis coloration. A complementation test of the pe mutant with the TALEN-mediated Bm-cardinal-deficient strain showed that the mutant phenotype could not be rescued, indicating that Bm-cardinal is responsible for pe. Moreover, knockdown of the cardinal homolog in Tribolium castaneum also induced red compound eyes. Our results indicate that cardinal plays a major role in ommochrome synthesis of holometabolous insects.

  16. Green cocoons in silkworm Bombyx mori resulting from the quercetin 5-O-glucosyltransferase of UGT86, is an evolved response to dietary toxins.

    Science.gov (United States)

    Xu, Xu; Wang, Meng; Wang, Ying; Sima, Yanghu; Zhang, Dayan; Li, Juan; Yin, Weiming; Xu, Shiqing

    2013-05-01

    The glycosylation of UDP-glucosyltransferases (UGTs) is of great importance in the control and elimination of both endogenous and exogenous toxins. Bm-UGT10286 (UGT86) is the sole provider of UGT activity against the 5-O position of quercetin and directly influences the formation of green pigment in the Bombyx cocoon. To evaluate whether cocoon coloration evolved for mimetic purposes, we concentrated on the expression pattern of Ugt86 and the activities of the enzyme substrates. The expression of Ugt86 was not only detected in the cocoon absorbing and accumulating tissues such as the digestive tube and silk glands, but also in quantity in the detoxification tissues of the malpighian tubes and fat body, as well as in the gonads. As in the green cocoon strains, Ugt86 was clearly expressed in the yellow and white cocoon strains. In vitro, the fusion protein of UGT86 showed quercetin metabolic activity. Nevertheless, Ugt86 expression of 5th instar larvae was not up-regulated in the silk gland by exogenous quercetin. However, it was significantly up-regulated in the digestive tube and gonads (P rutin, an insect resistance inducer and growth inhibitor typically found in plants, and to 20-hydroxylecdysone (20E), an insect endocrine and plant source hormone. On the contrary, up-regulated Ugt86 expression was almost nil in larvae exposed to juvenile hormone III (P > 0.05). The results of HPLC revealed that a new substance was formed by mixing 20E with the recombinant UGT86 protein in vitro, indicating that the effect of Ugt86 on 20E was similar to that on exogenous quercetin derived from plant food, and that the effect probably initiated the detoxification reaction against rutin. The conclusion is that the reaction of Ugt86 on the silkworm cocoon pigment quercetin is not the result of active mimetic ecogenesis, but derives from the detoxification of UGTs.

  17. 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

  18. 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.

  19. 家蚕的消化管与桑叶的协同进化研究%The Coevolution Study of Digestive Tract and Mulberry Leaf in Bombyx Mori

    Institute of Scientific and Technical Information of China (English)

    杨兰英; 刘再群; 王子茹; 杨汾芬; 吴文青

    2011-01-01

    With silver staining to stain the Bombyx mori, the results show that mulberry leaves in Bombyx mori's digestive tract include epidermis, mesophyll and vein. Epidermis includes upper epidermis and lower epidermis. Mesophyll includes palisade tissue and spongy parenchyma. Upper epidermis cells can be divided into three types: eystolith cells, green epidermal cells and yellow epidermal cells. There are stomas in lower epidermis. Mesophyll tissue includes crystals, most of them are in the spongy tissue. The digestive tract of Bombyx mori, can be divided into foregut, midgut and hindgut from fl'ont to back; Also, it can be divided into muscularis, basement membrane, epithelium and intima from outside to inside. One of the most developments is midgut. The developed epithelial cells of midgut go inside surface to make many large finger-shaped folds of prominency. Epithelial cells can be divided into two types: cylindrical cell, gobeh cell and regenerative cell. There are some differences in their shapes, functions and argyrophilics. Bombyx mori's digestive tract have different absorption efficiencies on the different organizations of mulberry leaves: the upper epidermis's is the first, the lower epidermis and palisade tissue's are the second, the spongy tissue's is the third. We used a method of staining animal cells to apply on plant cells and compared it with routine botany staining methods. Cells in upper epidermis of mulberry leaves can be divided into two subtypes depend on ditference of cells" argyrophilic.%通过银染法对家蚕整体染色,结果表明:家蚕消化管内的桑叶由叶表皮、叶肉和叶脉组成。叶表皮包括上表皮和下表皮。上表皮细胞可分为三种:钟乳体细胞、绿色表皮细胞和黄色表皮细胞;下表皮内含有气孔;叶肉组织内含有晶体,其中海绵组织内的最多。家蚕消化管由前向后可分为前肠、中肠和后肠,由外向内依次为肌层、底膜、

  20. Prokaryotic Expression and Antibody Preparation of Troponin C Ⅲ (TnC Ⅲ)from Bombyx mori%家蚕肌钙蛋白CⅢ基因的原核表达及其抗体的制备

    Institute of Scientific and Technical Information of China (English)

    严巧灵; 陈剑清; 王丹; 叶曼; 吴祥甫; 张耀洲

    2010-01-01

    家蚕肌钙蛋白CⅢ(TnCⅢ)是肌钙蛋白C(TnC)的一种亚型结构,本研究利用大肠杆菌表达TnCⅢ融合蛋白,并制备其多克隆抗体.从本实验室的家蚕(Bombyx mori)蛹cDNA文库中搜索到1条编码家蚕肌钙蛋白CⅢ亚型的cDNA序列,将其命名为BmTnCⅢ(Bombyx mori TnCⅢ).该cDNA序列含有完整的ORF,可编码153个氨基酸残基组成的TnCⅢ蛋白.根据该cDNA序列,设计带有BamH Ⅰ和HindⅢ酶切位点的引物,通过RT-PCR的方法从家蚕蛹总RNA中扩增到BmTnCⅢ亚型基因的完整的ORF序列(GenBank登录号为DQ889211),并将其重组到原核表达载体pET-28a相应的酶切位点上.将获得的重组质粒pET-28a-BmTnCⅢ转化大肠杆菌(Escherichia coli)Rosetta(DE3),经1 mmol/L IPTG诱导表达重组蛋白后,在分子量约21 kD处出现融合蛋白条带.用亲和层析的方法纯化目的蛋白His-Tag BmTnCⅢ,并以此为抗原免疫雄性新西兰白兔(Oryctolagus cuniculus),制各了该融合蛋白的多克隆抗体.Western blot结果表明,该多克隆抗体具有较高的特异性,可用于后续的组织分布与定位研究.

  1. 家蚕丝氨酸蛋白酶基因BmHP21的克隆及表达分析%Molecular Cloning and Expression Analysis of Serine Protease Gene BmHP21 in Silkworm, Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    刘碧朗; 李玉欣; 亓希武; 向仲怀; 何宁佳

    2011-01-01

    丝氨酸蛋白酶参与昆虫对抗入侵病原体及保护受伤组织的黑化反应.为了探讨家蚕黑化反应过程中丝氨酸蛋白酶在酚氧化酶原前体级联体系的作用,克隆了家蚕的一个新的丝氨酸蛋白酶基因,命名为BmHP21(CenBank登录号:JF431073).该基因由1 233个核苷酸组成,编码410个氨基酸.通过SMART网站预测蛋白结构显示BmHP21包括一个clip 结构域和一个胰蛋白酶结构域.BmHP21蛋白和烟草天蛾Ms-HP21的氨基酸序列有55%的相似性,推测BmHP21在家蚕黑化反应中起着激活酚氧化酶原前体激酶(PPAE)的作用.RT-PCR检测结果表明,BmHP21在家蚕大部分组织包括脂肪体中都有表达,其mRNA转录几乎贯穿于整个家蚕的幼虫、蛹和成虫发育阶段,推测BmHP21除了参与黑化反应外,还有其它的生理功能.%Serine proteases are involved in melanization which is used to combat with the intrusive pathogens and protect the damaged tissue in insect. To explore the role of serine proteases in the cascade activating system of prophenoloxidase during melanization in silkworm ( Bombyx mori), we cloned a novel serine protease gene, designated as BmHP21 (GenBank accession No. JF431073). The full-length cDNA of BmHP21 is 1 233 bp, encoding a putative protein with 410 amino acids. Protein structure prediction on SMART website displayed that BmHP21 contains a clip domain and a trypsin domain. The amino acid sequence of BmHP21 shares 55% identity with that of Ms-HP21 in Manduca sexta, suggesting that BmHP21 might function in the activation of prophenoloxidase-activating enzyme during melanization in silkworm. RTPCR detection revealed that BmHP21 was expressed in almost all tissues including fat body. The developmental expression profile of BmHP21 indicated that BmHP21 mRNA was expressed constitutively in larval, pupal and adult stages,suggesting that, besides of involvement in melanization, BmHP21 has other physiological functions as well.

  2. Analysis of Two-Dimensional Gel Electrophoresis Images of Protein from Posterior Silk Gland of Silkworm (Bombyx mori) on Day 1 and Day 4 in the 5th Instar Stage

    Institute of Scientific and Technical Information of China (English)

    WU Wei-cheng; ZHONG Bo-xiong; GAO Qi-kang; CHEN Jin-e; YE Jian; QIAN Yang-wen; LI Jian-ying; LU Hua-yun; MENG Zhi-qi; NI Chun-xiao

    2007-01-01

    The posterior silk gland (PSG) of silkworm is an important organ where fibroin is synthesized and secreted exclusively.Because fibroin constitutes 75-80% of the silk filament, the mechanism governing fibroin secretion, quality and yield of cocoon can be elucidated by the study on the PSG. Using two-dimensional gel electrophoresis (2-DE) and image analysis system, the changes in the protein composition in the PSG cell were investigated on the day 1 (D1) and day 4 (D4) in the 5th instar stage from five different strains of silkworm (Bombyx mori). While differences at protein level between days and strains were far less than those observed at the gene level using EST analysis. The change trends in protein composition from D1 to D4 were diverse among the different strains. The results suggest that the secretion of fibroin is regulated by multiple proteins. The site of regulation and the proteins responsible for the regulation vary with the strain, which leads to differences between strains in the capacity of fibroin secretion in the PSG cell.

  3. 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

  4. 家蚕Aly/REF的基因克隆、序列分析及其细胞定位%Cloning, sequence analysis and cellular localization of Aly/REF from the silkworm, Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    钟金凤; 曹广力; 薛仁宇; 贡成良

    2011-01-01

    RNA和输出因子结合蛋白(RNA and exportfactor binding proteins,REF/Aly)参与RNA的稳定、加工和输出.本研究参照已公开的家蚕Bombyx mori aly序列(GenBank登录号:DQ497195.1),通过RT-PCR克隆了家蚕aly/ref基因(Bmaly/ref).测序结果显示,该基因的开放读码框为765 bp,编码254个氨基酸残基,BmAly/REF与黑腹果蝇Drosophila melanogaster、小鼠Mus musculus的同源体的氨基酸序列一致性分别为49.7%和52.7%.结构预测结果显示,BmAly/REF具有REF家族的与RNA结合的结构域RRM,N和C端分别具有REF-N和REF-C基序.进化分析结果显示,昆虫的ALY/REF聚为一类,BmAly/REF与赤拟谷盗tribolium castaneum、意大利蜜蜂Apis mellifera的Aly/REF较为接近.将Bmaly/ref基因克隆进pGS21a(+)载体进行原核表达,重组蛋白免疫小鼠,获得鼠抗BmAly/REF多抗.免疫荧光实验结果显示,BmAly/REF在细胞质和细胞核中均有分布,但主要分布在细胞核中.芯片数据分析显示Bmaly/ref基因在家蚕幼虫5龄第3天各组织中均有较高水平的表达.研究结果显示BmAly/REF可能在RNA的核输出方面发挥作用,为进一步探讨BmAly/REF的功能奠定了基础.%RNA and export factor binding proteins (Aly/REF) play important roles in RNA stability, processing and nuclear export. Referring to the sequence of Bombyx mori aly gene ( GenBank accession no. DQ497195.1 ), the aly/refgene (Bmaly/ref) from the silkworm, B. mori, was cloned via RT-PCR. The sequencing result showed that the open reading frame (ORF) of Bmaly/ref is 765 bp in length, encoding 254 amino acid residues, and the amino acid sequence of BmAIy/REF showed 49.7% and 52.7% identities with the homologs of Drosophila melanogaster and Mus musculus, respectively. The structure prediction displayed that BmAIy/REF contains RNA binding domain (RRM), REF-N and REF-C motif of REF subfamily. Phylogenetic analysis showed that the insect Aly/REF proteins clustered to a group and the BmAly/REF was close

  5. 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.

  6. 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

  7. Preliminary evaluation of some biparental progenies of two bivoltine silkworm (Bombyx mori L. ) hybrids%家蚕二化性杂交品种部分数量性状遗传参数的初步评估

    Institute of Scientific and Technical Information of China (English)

    Gulam Nabi MALIK; Syed Zia Ul Haque RUFAIE; Tejender Paul SINGH; Mohammed Farooq BAQUAL; Habib Ullah DAR

    2005-01-01

    Twenty two BIPs (biparental progenies) generated from two bivoltine silkworm ( Bombyx mori L. ) hybrids, viz., SH6 × NB4D2 and CSR2 × CSR4, were evaluated for their performance in seven metric traits (weight of 10 mature larvae, single cocoon weight, single shell weight, shell ratio (%), effective rate of rearing, yield/10 000 larvae (wt.) and filament length.The experiment was laid out in a completely randomized design (CRD) with three replications for each treatment. The aim was to short list promising progenies and to work out estimates of direct selection parameters like heritability and genetic advance so that the generated information is utilized for the formulation of effective breeding and selection procedures aimed at extraction of new and more productive genotypes. BIPs 2, 4, 5, 6, 7, 10, 14, 16, 19 and 20 displayed a significantly superior performance in several traits. Single cocoon weight, yield/10 000 larvae (wt.) and filament length exhibited moderate estimates of heritability and reasonable genetic gains indicating genotypic variability to be a major component of phenotypic variability; hence, scope exists to bring about improvement in these traits through simple phenotypic selection. The rest traits (weight of 10 mature larvae,single shell weight, shell ratio (%) and ERR) recorded moderate estimates of heritability and low genetic gains environmental variability to be a major component of phenotypic variability; hence, selection for these traits would be less effective.%采用完全随机设计法根据10头老熟幼虫体重、全茧重、茧层量、茧层率(%)、存活率、万蚕茧层量和茧丝长等指标,对两对二化性家蚕Bombyx mori L.杂交品系(SH6×NB4D2和CSR2×CSR4)杂交一代的22个子代个体进行了遗传参数估算,以缩小优质蚕品种的候选范围,并且计算出直接筛选的参数,如遗传力和遗传进度等,使这些信息可用于以筛选高产新品种为目的的育种

  8. 家蚕核糖体18S RNA基因的序列分析及分子系统学研究%Sequence of the 18S Ribosomal RNA Gene of Ofsilkworm (Bombyx mori) and Molecular Systematics Analysis

    Institute of Scientific and Technical Information of China (English)

    魏国清; 代君君; 刘朝良; 张和禹

    2006-01-01

    为研究家蚕18S DNA基因的特点及分子进化,以家蚕丝腺为材料提取家蚕基因组DNA,通过PCR扩增、测序鳞翅目家蚕(Bombyx mori)核糖体小亚基18S RNA基因(18S rDNA)的全序列,将该序列与等翅目、直翅目、(衤责)翅目、鞘翅目、膜翅目、双翅目、捻翅目、弹尾目、蜉蝣目各一种昆虫的18S rDNA进行了比较.用DNAstav软件分析并进行序列比对,结果表明,昆虫18S rDNA有4段序列较为保守,以18S rDNA比对的第2保守区段构建的分子系统发育树表明鳞翅目和双翅目、膜翅目、鞘翅目进化关系最近,等翅目、直翅目、(衤责)翅目进化上较为接近,捻翅目昆虫与弹尾目昆虫的亲缘关系较为接近,捻翅目在分类上应是一种古老的昆虫.

  9. Cloning of Adenine Nucleotide Translocase Gene From Bombyx mori Based on EST Database%利用EST库资源克隆家蚕腺苷酸转移酶基因

    Institute of Scientific and Technical Information of China (English)

    陈大福; 牛宝龙; 翁宏飚; 孟智启; 吕顺霖

    2004-01-01

    根据物种间同源基因相对保守的特点,利用生物信息学方法以果蝇(Drosophila melanogaster)腺苷酸转移酶基因(adenine nucleotide translocase, ant) cDNA序列作为模板,对家蚕(Bombyx mori) EST数据库进行同源检索筛选,克隆了家蚕腺苷酸转移酶基因的cDNA序列(GenBank登录号为AY227000),全长为1 936 bp,并经RT-PCR克隆、序列分析验证,结果与电子克隆序列完全一致.该cDNA序列具有完整的开放阅读框架(ORF, 207~1 109 bp),推测编码蛋白为300个氨基酸,通过与烟草天蛾(Manduca sexta)、蜜蜂(Apis mellifera)、绿蝇(Lucilia cuprina)、果蝇、蚊子(Anopheles gambiae)等昆虫的腺苷酸转移酶蛋白序列比较,发现该基因具有高度的保守性.表明根据物种间同源基因序列,对跨物种间EST数据库进行同源检索筛选、拼接,是基因克隆的一条有效途径.

  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. 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.

  12. 家蚕BmE(spl)-like基因的克隆与表达谱分析%Cloning and Expression Profile Analysis of BmE(spl)-like Gene in Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    刘颖硕; 陆旋; 聂作明; 王丹; 陈健; 吕正兵; 张耀洲

    2012-01-01

    从本实验室构建的家蚕蛹cDNA文库中筛选到一条新的cDNA序列,生物信息学分析发现其编码的蛋白质序列与果蝇Enhancer of split基因[E(spl)]编码的DNA结合转录调控因子bHLH(helix-loop-helix)有较高的同源性,将该基因命名为BmE(spl)-like(Bombyx mori Enhancer of split like).该基因的ORF长606 bp,编码201个氨基酸残基,预测蛋白质分子质量22.3 kD,等电点9.7 .构建重组表达质粒pET-28a-BmE(spl)-like并在E.coli BL21(DE3)菌株中表达BmE(spl)-like蛋白,经亲和层析和FPLC反向柱纯化融合蛋白His-BmE(spl)-like并制备了抗体效价1∶25600以上的多克隆抗体.用qRT-PCR方法检测BmE(spl)-like在家蚕成虫发育期的转录水平最高,结合果蝇同源基因的功能,推测该基因与蚕蛾的翅发育相关;该基因在家蚕5龄幼虫各组织中的转录水平以表皮最高,马氏管中最低.Western blotting检测BmE(spl)-like在家蚕5龄幼虫性腺中的表达水平最高,其次是丝腺、表皮、脂肪体、肠组织,在头部、马氏管和气管中的表达水平极低,与qRT-PCR检测的BmE(spl)-like在各组织中的转录水平有些差异.%A novel cDNA sequence was screened from the silkworm pupa cDNA library constructed in our laboratory. Bioinformatics analysis showed that the protein sequence encoded by this gene shares a high homology with the inhibitor of DNA binding bHLH ( basic helix-loop-helix) encoded by Enhancer of split gene [ E( spl) ] of Drosophila melanogaster, and was thus designated as BmE{ spl) -like ( Bombyx mori Enhancer of split like). It contains an ORF of 606 bp and encodes a protein of 201 amino acids with a predicted molecular mass of 22. 3 kD and an isoelectric point of 9. 7. We con-structed the recombinant expression plasmid pET-28a-SmE( spl) -like and expressed BmE (spl) -like protein in E. coli BL21(DE3). After purification using affinity chromatography and FPLC reversed-phase column, the expressed fusion protein His

  13. 家蚕磷酸吡哆醛激酶cDNA的克隆和酶活表征%Bombyx mori Pyridoxal Kinase cDNA Cloning and Enzymatic Characterization

    Institute of Scientific and Technical Information of China (English)

    石瑞君; 张剑韵; 江昌俊; 黄龙全

    2007-01-01

    Pyridoxal kinase (PLK) (EC 2.7.1.35) catalyzes the ATP-dependent phosphorylation of pyridoxal, generating pyridoxal-5'-phosphate (PLP), an important cofactor for many enzymatic reactions. Bombyx mori, similar to mammals, relies on a nutritional source of vitamin B6 to synthesize PLP. This article describes how a cDNA encoding PLK was cloned from Bombyx mori using the PCR method (GenBank accession number: DQ452397). The cDNA has an 894 bp open reading frame and encodes a protein of 298 amino acid residues with a molecular mass of 33.1 kDa. The amino acid sequence shares 48.6% identity with that of human PLK, and it also contains signature conserved motifs of the PLK family. However, the protein is 10 or more amino acids shorter than the PLK from mammals and plants, and several amino acid residues conserved in the PLK from mammals and plants are changed in the protein. The cDNA cloned was expressed successfully in Escherichia coli using the T7 promoter/T7 RNA polymerase expression system, and the crude extracts containing the expressed product were found to have strong PLK enzymatic activity with a value of 30 nmol/min/mg, confirming that the cDNA encodes the functional PLK of Bombyx mori. This is the first identification of a gene encoding PLK in insects.%吡哆醛激酶(EC 2.7.1.35)在ATP和Zn2+的存在下,催化吡哆醛的磷酸化反应生成磷酸吡哆醛(PLP).在生物体内许多酶促反应中,PLP是一种重要的辅酶因子.家蚕和哺乳动物一样,需依赖食物中的维生素B6前体来合成PLP.文章描述了利用家蚕基因组数据库序列信息及使用PCR方法,克隆出编码家蚕吡哆醛激酶的cDNA(GenBank登录号:DQ452397).克隆到的cDNA含有一个894 bp的完整可读框,编码一条分子量为33.1 kDa,含298个氨基酸残基的蛋白质.序列比对显示此蛋白质序列与人类吡哆醛激酶蛋白序列具有48.6%的同一性,包含吡哆醛激酶家族共有的特征保守序列,但其氨基酸残基数比哺乳

  14. 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...

  15. 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...

  16. 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

  17. 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

  18. 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

  19. Molecular Cloning and Sequence Analysis of Glutamate Transporter Genes in Bombyx mori%家蚕谷氨酸转运蛋白基因克隆及序列分析

    Institute of Scientific and Technical Information of China (English)

    王晓莹; 袁刚祥; 齐登伟; 马艳; 沈以红

    2011-01-01

    Glutamate transporters are mainly distributed in the nerve, neuron and glial cells on the serosal. They regulate extracellular glutamate concentrations and play an important role in maintaining the normal physiological state of cells and in avoiding over-excitation of neurons. Blasting the silkworm genome with the glutamate transporter protein sequences in Trichoplusia ni and Drosophila melanogaster as querying probes, we obtained two homologous sequences (BmEAAT1 and BmEAAT2) in Bombyx mori, which were then cloned and sequenced. Both BmEAAT genes were shown to contain 8 exons and the deduced proteins were glycoproteins with 8 transmembrane domains each. Phylogenetic analysis showed that the two copies of BmEAAT genes in B. mori duplicated at least before divergence between Lepidoptera and Diptera. The information from microarray indicated that the expression profiles considerably differ between BmEAATl and BmEAAT2 in 10 tissues on the third day of the 5th instar silkworm, suggesting that function differentiation may have occurred between the two gene copies.%谷氨酸转运蛋白( EAAT)主要存在于神经、神经原及神经胶质细胞浆膜上,对胞外谷氨酸浓度进行调节,在维持细胞正常的生理状态、避免神经元过度兴奋方面起着重要的作用.利用鳞翅目粉纹夜蛾(Trichoplusia ni)和双翅目拟暗果蝇(Drosophila pseudoobscura)的谷氨酸转运蛋白序列为问询序列,在家蚕基因组数据库中进行同源检索,找到2个EAAT基因的同源基因(Bm EAAT),并进行了克隆测序.两个BmEAAT基因均含8个外显子,编码蛋白均为膜蛋白,各有8个跨膜结构域.多物种的同源基因序列的系统发生分析结果表明:家蚕的两个EAAT拷贝至少在鳞翅目与双翅目分化前就已存在.基因芯片数据显示Bm EAAT的两个拷贝在家蚕5龄3天的组织表达模式有明显差异,表明这两个基因的功能可能已经分化.

  20. 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.

  1. 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

  2. 荧光茧色判性家蚕黄酮合酶I(BmFNS I)基因在家蚕组织内的表达差异%Expression Differences of Flavonoid Synthase I ( BmFNS I) in the Tissues of Bombyx Mori with Sex-identified Fluorescent Cocoon Color

    Institute of Scientific and Technical Information of China (English)

    许乃霞; 张雨青; 沈卫德

    2011-01-01

    采用荧光定量PCR分别检测不同品种荧光判性家蚕雌、雄家蚕的中肠、丝腺和脂肪体中的黄酮合酶I(BmFNS I)基因的表达.结果表明,家蚕BmFNS I基因在各品种荧光判性家蚕中肠中的相对表达量最高,其次是脂肪体,在丝腺中的表达量较低,说明BmFNSI基因在对家蚕食物桑叶内的黄酮类化合物的合成代谢中起重要作用;但是在各品种荧光判性家蚕的雌、雄中肠中的表达并无明显差异,说明荧光判性家蚕雌、雄蚕在对桑叶内的黄酮类化合物的合成代谢途径上不存在差异,至少在FNS I基因所调节的黄酮类化合物的合成代谢上不存在差异.%Expression differences of Flavonoid synthase I ( BmFNS I) gene in the midgut, silk gland and fat body of the male and female Bombyx mori with sex-identified fluorescent cocoon color was detected by fluorescence quantitative PCR. And the result showed that the relative expression level BmFNS I gene in the midgut of the silkwormn was the highest, followed by that in fat body, and the expression level in silk gland was the lowest, indicating that BmFNS I gene played an important role in flavonoids anabolism in leaves of mulberry, which was the food of silkworm. However, there was no significant difference in the BmFNS I gene expression level in the midgut between male and female Bombyx mori varieties with sex-identified fluorescent cocoon color, suggesting that there was no difference in anabolism of flavonoids in leaves between the male and female Bombyx mori with sex-identified fluorescent cocoon color, at least no difference in anabolism of flavonoids regula ted by flavonoid synthase I gene.

  3. 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

  4. Activation of BmGSTd1 promoter and regulation by transcription factor Krüppel (Kr) in silkworm, Bombyx mori.

    Science.gov (United States)

    Zhao, Guodong; Wang, Binbin; Liu, Yunlei; Du, Jie; Li, Bing; Chen, Yuhua; Xu, Yaxiang; Shen, Weide; Xia, Qingyou; Wei, Zhengguo

    2014-11-10

    The Glutathione S-transferases (GSTs) are a large family of multifunctional enzymes, many of which play an important role in the detoxification of endogenous and exogenous toxic substances. In this research, firstly, we measured the rutin-induced transcriptional level of BmGSTd1 gene by using real-time quantitative RT-PCR method and dual spike-in strategy. The activities of the BmGSTd1 promoter in various tissues of silkworm were measured by firefly luciferase activity and normalized by the Renilla luciferase activity. Results showed that the activity of the BmGSTd1 promoter were highest in Malpighian tubule, followed by fat body, silk gland, hemocyte, epidermis, and midgut. The essential region for basal and rutin-induced transcriptional activity was -1573 to -931bp in Malpighian tubule and fat body of silkworm. Promoter truncation analysis using a dual-luciferase reporter assay in BmN cells showed that the region -1288 to -1202bp for BmGSTd1 gene was essential for basal and rutin-induced transcriptional activity. Sequence analysis of this region revealed several potential transcriptional regulatory elements such as Bcd and Kr. The mutation of core base of Kr site demonstrated that Kr functioned positively in rutin-mediated BmGSTd1 transcription.

  5. Analysis of reference gene expression for real-time PCR based on relative quantitation and dual spike-in strategy in the silkworm Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    Ran Peng; Yuanfen Zhai; Hua Ding; Tianyuan Di; Ting Zhang; Bing Li; Weide Shen; Zhengguo Wei

    2012-01-01

    In general,for real-time quantitative polymerase chain reaction (qPCR),normalization strategies use a reference gene as a control and to avoid the introduction of experi-mental errors expression of this gene should not vary in response to changing conditions.However,the expression of many reference genes has been reported to vary consid-erably and,without appropriate normalization,the expression profile of a target gene can be misinterpreted.In this study,the expression levels of seven commonly used reference genes (ACT,GAPDH,28srRNA,RPL3,α-tubulin,UBC,and TBP) were detected at different development time points and in response to treatment with 20-hydroxyecdysone (20E) and with rntin.The expression stability was analyzed using geNorm and NormFinder software.Significant variations were found among normal tissues and between experimentally treated tissues.The dual spike-in strategy also revealed significant variations of the expression levels of the reference genes among normal tissues and between experimentally treated tissues.Glutathione-S-transferase sigma 1 (GSTs1),which has a high expression level in fat body and is related to the mechanism of resistance,was used as a target gene to validate the feasibility and difference of these two approaches.

  6. 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

  7. 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

  8. 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

  9. 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

  10. 环境激素2,4-二氯苯酚对家蚕生殖发育的影响%Effects of 2,4-Dichlorophenoi on the Genital Development of Silkworm(Bombyx mori)

    Institute of Scientific and Technical Information of China (English)

    袁红霞; 裔洪根; 陈息林; 徐世清

    2009-01-01

    The aim of this study was to evaluate the estrogen effects of 2,4-dichlorophenol(2,4-DCP) on lepidopteron insect. The gonad growth and development of silkworm(Bombyx mori) were investigated by feeding with 2,4-DCP-added artificial feedstuffs. Silkworms were placed in culture boxes feeding with 2,4-DCP-added(0,0.2,0.4,0.8,1.6 and 3.2 mmol·kg~(-1)) artificial feedstuffs in a culture container kept at constant temperature and humidity in each instar, in which each treatment involved three boxes and each boxes received 50 silkworms. The gonad index, oocyte, spermatid and sperm numbers in fifth instars, pupas and moths were investigated. The egg production, egg deposition numbers and unfertilized egg rates were calculated. The results showed that, with treatment increasing, at 0.2~0.8 mmol·kg~(-1) 2,4-DCP, the o-vary growth and development of fifth instars and pupas were significantly promoted(P<0.05),11.6 mmol·kg~(-1) 2,4-DCP inhibited the ovary development of fifth instars in 168 h. However, ovary development in pupas were increased obviously. The oocyte development of fifth instars and pupas was elevated at high level. 2,4-DCP decreased the spermary development, especially at late development stages of fifth instars and pupas. The testicles of male moth was retrogressed at 1.6 mmol·kg~(-1) 2,4-DCP.2,4-DCP decreased the spermatid numbers of pupas and sperm numbers of moths significantly(P<0.01). At 1.6 mmol·kg~(-1) 2,4-DCP, the egg deposition decreased to 20 percent of control, however the egg production numbers were 1.3 times as much as control. At 0.8 mmol·kg~(-1) or higher, the unfertilized egg rate increased remarkably (P<0.05). The 2,4-DCP functioned as estrogen effects through inhibiting the growth and development of spermary and germ cells.%为了探讨环境激素对鳞翅目昆虫的影响,用添加2,4-二氯苯酚(2,4-DCP)的人工饲料饲养家蚕,调查2,4-DCP对家蚕生殖发育的影响.结果显示,1.60mmol·kg~(-1)以下浓度的2,4-DCP,对

  11. Using Different Staining Agents to Identify the Living Status of Circulating Hemocytes in the Silkworm, Bombyx mori%利用不同染料复合染色判定家蚕血细胞的存活状况

    Institute of Scientific and Technical Information of China (English)

    李旭全; 杨兵; 路岸瑞; 刘朝良; 凌尔军

    2011-01-01

    Bombyx mori has 5 types of circulating hemocytes that can be clearly identified after being stained by acridine orange (AO) and propidium iodide (PI). 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) is another type of staining agent that has been extensively used to study cell proliferation and to identify living cells. Bombyx mori hemocytes undergone multiple staining, I. E. Staining with MTT first and then with AO and PI, showed two patterns. One was that a majority of granulocytes and plasmatocytes and all prohemocytes could not be stained by MTT but were shown to be living cells from AO staining. The other was that the MTT stained hemocytes had a lot of purple fiber-like structure extending from cell surface. All MTT-stained hemocytes but some granulocytes with pseudopods could also be stained by PI according to their nuclei with red fluorescence. Therefore, careful analysis and discriminative treatment should be conducted when multiple staining with these three staining agents are used to tell whether a cell is living or dead.%家蚕有5种血细胞,可以通过吖啶橙(acridine orange,AO)和碘化丙啶(propidium iodide,PI)染色来区分.溴化噻唑蓝四氮唑[3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2 H-tetrazolium bromide,MTT]是一种广泛应用于细胞增殖和活细胞鉴定的染色剂.通过对家蚕血细胞进行复染,即先用MTT染色,再用AO和PI染色,发现以下2种情况:很多颗粒细胞、浆细胞和所有的原血球细胞不能被MTT染色,但AO染色显示为活的细胞;被MTT染色的细胞表面有紫色纤维状物伸出,除了一部分具有伪足的颗粒细胞外,所有被MTT染色的血细胞也被PI染色,细胞核是红色.因此,利用这3种染料对家蚕血细胞复合染色来判断细胞的存活与死亡,需要仔细分析并分别对待.

  12. Functional analysis of Bombyx Wnt1 during embryogenesis using the CRISPR/Cas9 system.

    Science.gov (United States)

    Zhang, Zhongjie; Aslam, Abu F M; Liu, Xiaojing; Li, Muwang; Huang, Yongping; Tan, Anjiang

    2015-08-01

    Recently established, custom-designed nuclease technologies such as the clustered regularly interspaced short palindromic repeat (CRISPR)-associated system provide attractive genome editing tools. Targeted gene mutagenesis using the CRISPR/Cas9 system has been achieved in several orders of insects. However, outside of studies on Drosophila melanogaster and the lepidopteron model insect Bombyx mori, little success has been reported, which is largely due to a lack of effective genetic manipulation tools that can be used in other insect orders. To create a simple and effective method of gene knockout analysis, especially for dissecting gene functioning during insect embryogenesis, we performed a functional analysis of the Bombyx Wnt1 (BmWnt1) gene using Cas9/sgRNA-mediated gene mutagenesis. The Wnt1 gene is required for embryonic patterning in various organisms, and its crucial roles during embryogenesis have been demonstrated in several insect orders. Direct injection of Cas9 mRNA and BmWnt1-specific sgRNA into Bombyx embryos induced a typical Wnt-deficient phenotype: injected embryos could not hatch and exhibited severe defects in body segmentation and pigmentation in a dose-dependent manner. Quantitative real-time PCR (qRT-PCR) analysis revealed that Hox genes were down-regulated after BmWnt1 depletion. Furthermore, large deletion, up to 18Kb, ware generated. The current study demonstrates that using the CRISPR/Cas9 system is a promising approach to achieve targeted gene mutagenesis during insect embryogenesis.

  13. 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...

  14. 家蚕肠道产蛋白酶菌株的分离与鉴定及其发酵条件%Isolation and Identification of the Protease-producing Bacteria from Gut of Silkworm (Bombyx mori) and Its Suitable Fermentation Condition

    Institute of Scientific and Technical Information of China (English)

    王在贵; 杨文静; 刘朝良; 朱保建; 魏国清; 邹昌瑞

    2011-01-01

    To explore the species distribution and probiotics function of enzyme-producing bacteria in gut of the silkworm, protease-producing bacteria strains were isolated from the inclusion in 4th larval guts of the silkworm Bombyx mori by NA mediums and casein-NA mediums and 16 Sr DNA sequences were used to identify the species.Meanwhile, the enzyme producing activities of the three strains and optimum fermentation conditions of Haoyue NA1 strain were investigated.The results showed that three strains Haoyue NA1,951 NA3 and 951 NA6 belonged to Acinetobacter were isolated and confirmed.The maximum protease activity with 29.5 U/mL was found in Haoyue NA1 and the optimum fermentation conditions were com powder 10 000 mg/L, soybean powder 10 000 mg/L, MgSO4 400 mg/L, NaC1 15 000 mg/L and K2HPO4 1 000 mg/L with a liquid volume of 80 mL in 150 mL bottol and cultured at 35 ℃, pH 9.0 under 180 r/min for 48 h.In this condition, the maximum protease activity produced by Haoyue NA1 reached 50 U/mL.This research can play an important role on the research of distribution ofmicrorganisms in gut of the silkworm and on the control of the gut probiotics function.%为探明家蚕肠道中产蛋白酶细菌的种类分布及其作用效果,本研究以家蚕(Bombyx mori)4龄幼虫肠道内容物为材料,采用牛肉膏蛋白胨培养基和酪蛋白培养基分离筛选产蛋白酶细菌菌株,利用16SrDNA序列分析鉴定其种属,并研究其产酶能力和产酶活力较高菌株的最适发酵条件.结果获得3株产蛋白酶菌株皓月NA1、951 NA3和951 NA6,均归属于不动杆菌属(Acinetobacter),其中皓月NA1号细菌产酶能力最强,最佳产酶量为29.5 U/mL.以玉米粉10 000 mg/L、黄豆粉10 000 mg/L、MgSO4 400 mg/L、NaCl 15 000 mg/L和K2HPO4 1 000mg/L作为发酵培养基成分,在35℃、起始 pH9.0、装液量为80mL/150mL、180 r/min振荡培养48 h的优化发酵条件下,皓月NA1号细菌最大产酶量可达50 U/m L.研究结果对家蚕肠道微生

  15. 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

  16. 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

  17. 不同球孢白僵菌菌株生物学特性与其对家蚕致病力的关系%Relationship between Biological Characteristics of Beauveria bassiana ( Bals.) Vuill and Pathogenicity to Bombyx mori L.

    Institute of Scientific and Technical Information of China (English)

    骆海玉; 邓业成; 廖永梅; 李瑞钰

    2012-01-01

    [目的]探讨球孢白僵菌菌株的生物学特性与其对家蚕致病力的关系,为家蚕白僵病的防治提供科学依据.[方法]从我国各地收集6个白僵菌杀虫剂,并从广西养蚕区采集3个僵蚕样品,对从中分离纯化的菌株进行形态观察和分子鉴定,同时对家蚕进行致病力测定,比较各菌株菌落直径、产孢量、胞外蛋白酶产生水平等生物学特征指标的差异.[结果]获得的9个分离菌株均为球孢白僵菌(Beauveria bassiana Vuillemin);各菌株对家蚕都有较强致病力,且致病力有一定差异;各菌株菌落生长直径、产孢量及胞外蛋白酶活性均存在较大差异,并与其对家蚕的致病力之间存在相关关系.[结论]各菌株产孢量和胞外蛋白酶活性与其对家蚕的致病力之间存在显著正相关.%[Objective] This study was to investigate the relationship between biological characteristics of Beauveria bassiana ( Bals. ) Yuill and pathogenicity of Bombyx mori 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 silkworms to B. bassaina was determined, and the biological characteristics such as growth diameter, spore production amount and the extracellular protease activity of the different B. bassiana strains were compared. [ Result] The isolated 9 strains were al] B. bassaina (Bals. ) Vuillemin, and all strains had high pathogenicity to silkworm, but with different pathogenicities. The growth diameter, spore production amount and extracellular protease activity of different B. bassiana strains were also different, and showed correlation with the patheogenicity to silkworms

  18. 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.

  19. 家蚕和野桑蚕脂肪酸脱氢酶desat4全长cDNA和启动子的克隆及其原核表达%Cloning of full-length cDNA and promoter sequences of fatty acid desaturase gene desat4 from silkworms, Bombyx mori and B.mandarina,and its prokaryotic expression

    Institute of Scientific and Technical Information of China (English)

    陈全梅; 程道军; 马振刚; 胡晓明; 查幸福; 赵萍

    2012-01-01

    [目的]获得家蚕Bombyx mori和野桑蚕Bombyx mandarina脂肪酸脱氢酶基因desat4的cDNA及其启动子序列,并应用原核表达系统获得目的蛋白质,为进一步研究该基因的功能提供基础.[方法]利用RACE技术克隆家蚕和野桑蚕desat4基因全长cDNA序列,基于家蚕全基因组序列设计引物克隆它们的启动子,并采用原核表达技术对该基因进行异源表达.[结果]克隆得到家蚕与野桑蚕desat4基因全长cDNA,长度分别为1 717 bp和1 718bp,ORF长度为1 059 bp,编码352个氨基酸残基,结构上有3个组氨酸保守区和4次跨膜结构域,说明该蛋白质是膜蛋白.同源性分析发现Desat4氨基酸序列与烟草天蛾Manduca sexta MsexKPSE脱氢酶拥有88.9%相似性.克隆获得的家蚕和野桑蚕desat4基因启动子序列长度分别为1 808 bp和870 bp,该区域包含有热休克因子HSF(heat shock factor)结合位点、NIT2结合位点和CdxA(caudal type homeobox transcription factor A)结合位点等等,并未发现有典型的TATA-box,但在靠近5'-UTR区域发现了转录起始因子特征序列.时期表达谱分析显示desat4基因从刚产下卵到成虫(蛾)的36个不同发育时间点都有持续稳定的表达.应用原核表达系统成功地表达了Desat4蛋白质,并用温和变性剂十二烷基-β-D-麦芽吡喃糖苷( dodecyl-β-D-maltoside,DDM)就能很好地促进该蛋白质的溶解.[结论]本研究成功地克隆了家蚕和野桑蚕desat4基因全长cDNA及其启动子序列并进行了序列比对分析,构建了desat4基因的原核表达载体并成功表达,为进一步研究该基因的功能提供参考.%[ Aim ] The main aim of this research was to clone and align the full-length cDNA and promoter sequences of desatA gene from silkworms, Bombyx mori and B. mandarina, and construct prokaryotic expression vector to obtain a membrane protein Desat4 for further functional study. [ Methods ] Full-length cDNA of the desatA gene from silkworm was

  20. 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...

  1. 家蚕乳糖酶-根皮苷水解酶基因BmLPH014192的表达和序列选择性剪接分析%Expression and Sequence Alternative Splicing Analysis of the Lactase-phlorizin Hydrolase Gene BmLPH014192 in Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    黄飞飞; 赵小靓; 鲁成

    2013-01-01

    Lactase-phlorizin hydrolase (LPH) is a giycoprotein in microvillus membrane of the intestinal epithelial cell. It has hydrolytic activity to lactose and phlorizin. We identified one of the silkworm (Bombyx mori) LPH genes and named it as BmLPH014192. CDS (coding sequence) of this gene is 1 305 bp long and encodes 434 amino acids, with predicted molecular weight of 50. 8 kD and isoelectric point of 5. 35. An alignment of the cDNA sequence with silkworm genom-ic sequence revealed that there are 7 exons in BmLPH 014192 and all of the exon/intron boundaries conformed to the GT-AG rule. Sequence comparison between BmLPH 014192 and LPH genes from other species including Homo sapiens, Rattus norvegicus and Oryctolagus cuniculus of mammal and Drosophila melanogaster, Anopheles gambiae, Chilo suppressalis, Camponotus floridanus and Danaus plexippus of insect revealed that their amino acids all shared about 60% similarity. Phylogenetic analysis showed that BmLPH014192 was clustered with other insect LPH genes. Microarray analyzing results indicated that BmLPHO14192 was expressed only in midgut among 9 tissues of day 3 silkworm larvae of the 5th instar, and the expression level was very high. Fragments amplified from midgut cDNAs of Dazao, a variety spinning green cocoons, and of 19-710, a variety spinning white cocoons, were 1 000 and 750 bp long respectively, indicating their origination from alternative splicing. No specific band was amplified from midgut of other white cocoon species. It is suggested that the special expression pattern of BmLPH 014192 is related to the specific function of this gene in midgut tissues of different silkworm varieties.%乳糖酶-根皮苷水解酶(lactase-phlorizin hydrolase,LPH)是肠组织上皮细胞微绒膜上的一种糖蛋白,对乳糖和根皮苷有水解活性.鉴定了家蚕基因组中的一个LPH基因,命名为BmLPH014192.该基因CDS长1 305 bp,编码434个氨基酸,预测蛋白质分子质量为50.8 kD,等电点(pI)为5.35.

  2. 家蚕乙醇脱氢酶基因的表达特征及乙醇在蚕体内的代谢分析%Expressional Profile of Alcohol Dehydrogenase Genes and Metabolic Analysis of Ethanol in Bombyx mori Larvae

    Institute of Scientific and Technical Information of China (English)

    杜丽; 王长春; 徐云敏; 李玉欣; 何宁佳

    2012-01-01

    Alcohol dehydrogenase (ADH) is a critical ethanol metabolic enzyme in organisms. Bioinformatics analysis showed that there are 7 ADH coding genes in the silkworm (Bombyx mori) genome ( BmADH1 ~ BmADH7). Semi-quantitative RT-PCR indicated that BmADH2, BmADH3, BmADHA and BmADH5 had high expression level in silk gland of day 3 silkworm larvae of the 5th instar and BmADH1 , BmADH6 and BmADH7 had high expression level in fat body. After day 3 silkworm larvae of the 5th instar were treated with 28% or 56% ethanol via direct injection and oral feeding, the metabolism of ethanol in silkworm larvae as well as the variations of ADH gene expression and ADH enzyme activity in fat body were investigated. Semi-quantitative RT-PCR analysis indicated that the expressions of BmADH1, BmADH6 and BmADH1 genes were up-regulated in silkworm fat body after being treated by direct injection of 56% ethanol, while the expressions of these three genes remained unchanged after treatment with 28% ethanol. Enzyme activity assay revealed that ADH enzyme activities were predominantly increased in silkworm fat body at 1 h after treatment with 28% or 56% ethanol (P<0. 05). Gas chromatography analysis showed that ethanol was quickly converted into acetaldehyde in the larval hemolymph. These results indicate that, after silkworm larvae receive stimulation of high concentration ethanol, the expression of ADH genes is up-regulated in fat body, and the increased ADH enzyme activities participate in ethanol metabolic process to protect the larvae from being harmed by high concentration ethanol.%乙醇脱氢酶(alcohol dehydrogenase,ADH)是生物体内重要的乙醇代谢酶.生物信息学分析显示家蚕基因组中存在7个ADH编码基因(BmADH1 ~ BmADH7),半定量RT-PCR检测BmADH2、BmADH3、BmADH4和BmADH5在家蚕5龄第3天幼虫的丝腺中表达水平较高,BmADH1、BmADH6、BmADH7在脂肪体中高水平表达.利用直接注射和口器灌喂2种方式,对家蚕5龄第3

  3. Sequence Feature Analysis, Expression Pattern and Prokaryotic Expression of Defensin Genes from Silkworm ( Bombyx mori)%家蚕防御素基因的序列特征与表达模式分析及原核表达试验

    Institute of Scientific and Technical Information of China (English)

    马振刚; 贾俊杰; 陈全梅; 黄为; 李田; 潘国庆; 周泽扬

    2012-01-01

    Defensins of silkworm (Bombyx mori) are major members of silkworm antimicrobial peptide (AMP) family and important effectors of silkworm innate immune system.Bioinformatic analysis showed that two silkworm defensin genes,termed BmdefA and BmdefB,are located on chromosome 4 and chromosome 13 respectively.Each of them has two ex-ons.Prediction on isoelectric point showed that BmdefA brings negative charges,belonging to the unusual anionic peptide family,whereas BmdefB brings positive charges,belonging to the usual cationic peptide family.Molecular masses of mature peptides encoded by both genes are predicted to be around 4 kD.RT-PCR assay revealed that BmdefA was expressed during all developmental stages of silkworm and in all observed tissues of silkworm larvae and adults; BmdefB was expressed from day 3 of the 5th instar to adult stage with higher expression in male individuals over female ones and high expression levels in gonad,fat body and hemocyte of day 3 larvae of the 5th instar.Silkworm defensins BmdefA and BmdefB had different molecular characteristics and expression patterns,suggesting that they play different roles in the innate immune process of silkworm and even function via different antimicrobial mechanisms.After prokaryotic expression vector for fusion expression of silkworm defensin genes with GST tag was construe ted,induced expression in E.coli yielded soluble target proteins,which were further purified using GST affinity chromatography and identified by Western blotting.The present study establishes a good basis for further investigations on in vitro activity and antimicrobial mechanism of BmdefA and BmdefB.%家蚕防御素(defensin)是家蚕抗菌肽家族的主要成员之一,为家蚕先天性免疫的重要效应因子.采用生物信息学方法分析家蚕防御素基因BmdefA、BmdefB的序列中各有2个外显子,2个基因分别定位于家蚕第4号和第13号染色体上;等电点预测BmdefA带有负电荷,属于罕见的阴离子

  4. 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

  5. BmNPV or f98对家蚕核型多角体杆状病毒复制、转录及包装的影响%Influence of BmNPV orf98 on DNA replication,transcription and virus package of Bombyx mori nucleopolyhedrovirus

    Institute of Scientific and Technical Information of China (English)

    史利利; 蒋彩英; 于威; 陈琛; 蒋磊; 巩成见; 童富淡

    2015-01-01

    为了研究家蚕核型多角体病毒( Bombyx mori nucleopolyhedrovirus ,BmNPV)基因 orf98的功能,通过λRed重组系统定点敲除BmNPV or f98基因,构建缺失型重组病毒 Bm98‐ko‐Bacmid;以Bac‐to‐Bac系统补回BmNPV or f98基因,构建补回型重组病毒 Bm98‐re‐Bacmid;将野生型病毒( w tBacmid)、缺失型病毒( Bm98‐ko‐Bacmid)和补回型病毒( Bm98‐re‐Bacmid)分别转染家蚕细胞BmN .病毒滴度检测结果显示,Bm98‐ko‐Bacmid可形成侵染性的病毒粒子,但数量显著降低( P<0.05).透射电子显微镜观察发现,Bm98‐ko‐Bacmid只产生游离的杆状病毒粒子,数量明显减少,而w tBacmid和 Bm98‐re‐Bacmid产生大量具有囊膜结构的成熟病毒粒子.荧光定量聚合酶链反应分析结果表明,BmNPV or f98基因缺失对BmNPV病毒复制没有影响,而早期基因 le f3、晚期基因 v p39和极晚期基因p10的转录水平显著降低( P<0.05).综上所述,BmNPV or f98基因对病毒复制是非必需的,但显著影响病毒的繁殖速度和包装( P<0.05);对病毒各个时期的基因转录也具有重要影响.%Summary Baculoviruses have been considered as the powerful vectors to express the exogenous gene . And the representative vectors in baculovirus expression vector system is Autographa californica multinucleocapsid nucleopolyhedrovirus (AcMNPV) and Bombyx mori nucleopolyhedrovirus (BmNPV) . The AcMNPV expression system has been widely applied in American and European countries . However , the BmNPV expression reaches a higher level over other systems , because BmNPV can infect silkworm larva or pupa . Moreover , silkworm is pretty normal in China , with lower cost and mature breeding technology , thus it is really popular to use the silkworm as a biofactory" to produce recombinant protein . The BmNPV genome sequenced in 1999 was 128 413 nucleotides long with a G + C content of 40% and contained about 136 open reading frames ( ORFs) encoding predicted

  6. 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

  7. 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.

  8. 家蚕microRNA 7靶基因Bmhairy的鉴定和转录表达模式%Identification of Bmhairy as the Target of bmo-miR-7 and Its Transcriptional Expression Profiles in the Silkworm (Bombyx mori)

    Institute of Scientific and Technical Information of China (English)

    刘仕平; 吴小燕; 张丹宇; 黄亚玺; 王伟; 赵萍; 夏庆友

    2016-01-01

    [目的]microRNA是一类长约22个碱基的非编码RNA,通过与其靶基因的特异性结合来调控新陈代谢和生长发育等多种生命活动.鉴定家蚕(Bombyx mori) Bmhairy,比较Bmhairy和bmo-miR-7的表达模式,验证Bmhairy是否为bmo-miR-7的靶基因,为研究家蚕的变态发育机制提供线索.[方法]用家蚕胚胎反转期的总RNA反转录合成的cDNA模板,克隆Bmhairy编码区(coding DNA sequence,CDS)和3'端非翻译区(3'untranslated region,3'UTR)并进行序列分析.基于荧光定量PCR和芯片数据比较bmo-miR-7和Bmhairy的表达模式.利用RNAhybrid和MiRTif,预测和分析Bmhairy的mRNA 3'UTR上bmo-miR-7的靶位点.构建荧光素酶报告基因载体,用家蚕胚胎细胞系(BmE)进行共转染试验,验证bmo-miR-7对Bmhairy的 3'UTR结合位点.[结果]在家蚕中克隆了Bmhairy,含2个内含子和3个外显子,CDS长654 bp,编码217个氨基酸.Bmhairy的mRNA YUTR长366nt.Bmhairy高度保守,含有bHLH、ORANGE和WRPW结构域.Bmhairy与鳞翅目(Lepidoptera)蛱蝶科(Nymphalidae)中的黑脉金斑蝶(Danaus plexippus)相似度最高.Bmo-miR-7在家蚕胚胎期高量表达,在5龄第3天的精巢中相对较高,而Bmhairy在成虫期的表达量最高,在5龄第3天的头部相对较高.Bmhairy mRNh YUTR上有bmo-miR-7一个靶位点.共转染试验表明,bmo-miR-7下调Bmhairy.[结论]Bmhairy的家蚕时期表达和5龄第3天组织表达均与bmo-miR-7的表达呈相反的模式.靶基因预测和双荧光素酶转染试验表明Bmhairy是bmo-miR-7的靶基因,为进一步研究bmo-miR-7和Bmhairy在家蚕体内的生物学功能及调控关系提供了依据.

  9. Screening of a Cellulase-producing Strain from the Larval Gut of Bombyx mori and Optimum the Fermentation Condition%家蚕肠道纤维素酶菌株的筛选及产酶条件的优化研究

    Institute of Scientific and Technical Information of China (English)

    马如箭; 贾俊强; 刘艳伟; 任子旭; 桂仲争

    2013-01-01

    A novel cellulase-producing fungus named BMC-2 was isolated from the fifth-instar larval gut of Bom-byx mori. Fermentation conditions for the cellulase-producing, in which CMCase specific activity was taken as re-sponse value, were optimized with Plackett-Burman design, steepest ascent test and Box-Benhken design. The re-sults showed that fermentation time, fermentation temperature, initial pH of medium and rotational speed had influ-ence on cellulase-producing significantly. The affecting order was fermentation time, initial pH of medium, fermen-tation temperature and rotational speed, in turn. The optimized fermentation conditions were as follows:fermenta-tion time 94.35 h, fermentation temperature 30.3℃, initial pH of medium 7.01 and rotational speed 179 r/min. Under the optimal conditions, the CMCase specific activity was 25.801 U/mg in theory and 25.526 U/mg in actual value that was increased by 1 times compared with the initial fermentation condition. The result indicated that the established mathematical model by response surface methodology was feasible for practical prediction.%  从5龄家蚕肠道分离筛选得到一株产纤维素酶菌株BMC-2,以羧甲基纤维素酶(CMCase)比活力为响应值,通过Plackett-Burman试验设计、最陡爬坡试验和Box-Behnken试验设计对菌株BMC-2产纤维素酶发酵条件进行优化,结果表明,发酵时间、发酵温度、培养基初始pH和转速对CMCase比活力具有显著影响,其影响程度由大到小依次为发酵时间、培养基初始pH、发酵温度、转速.确定菌株BMC-2产纤维素酶最优发酵条件为:发酵时间94.35 h,发酵温度30.3℃,培养基初始pH 7.01,转速179 r/min.在此条件下, CMCase比活力理论值为25.801 U/mg,验证值为25.526 U/mg,较产酶条件优化前提高了1倍,预测模型可靠性高,可应用于菌株BMC-2产纤维素酶条件的优化.

  10. Effect of protection and refrigeration time alteration on hatching rate of acid-treated silkworm (Bombyx mori) eggs%不同库外保护时间和冷藏时间对冷藏浸酸种孵化率的影响

    Institute of Scientific and Technical Information of China (English)

    余武昌; 蒋满贵; 李国栋; 阳诚; 何珊珊; 莫云霞; 李燕飞

    2011-01-01

    [目的]进一步验证库外保护时间和冷藏时间对冷藏浸酸种孵化率的影响程度.[方法]以"两广二号"正、反交一代杂交种为研究对象,将不同库外保护时间(2~7 d,25℃)和不同内库冷藏时间(30~110 d,3~5℃)的蚕卵,经同一标准浸酸处理后统计其冷藏浸酸种孵化率.[结果]冷藏时间相同,库外保护时间短的冷藏浸酸种比库外保护时间长的发育快,而库外保护时间间隔越长,蚕种孵化整齐度越差.随着冷藏时间的延长,不同库外保护时间的冷藏浸酸种实用孵化率提高明显,冷藏时间达到70 d以上,其实用孵化率全部达到生产要求(90.0%以上);而且不同库外保护时间的冷藏浸酸种孵化整齐度也明显提高.[结论]冷藏浸酸种的卵龄差过大、内库冷藏时间不足等容易出现孵化不整齐.因此,在生产上应尽量减少冷藏浸酸种补种,防止蚕种孵化不整齐.%[Objective]The present experiment was aimed to investigate the effect of altering protection and refrigeration times outside storeroom on hatching rate of acid-treated silkworm (Bombyx mori) eggs. [Method]The hatching rate of acid-treated eggs (in HCl solution, 48℃), obtained from the cross and reciprocal cross species of silkworm variety Liangguang 2, was investigated after protecting them outside storeroom at 25℃ for 2-7 days and refrigerating at 3-5℃ for 30-110 days. [Result]The growth in the eggs kept for same refrigeration time was found faster in shorter protection time treated eggs compared to those kept for longer protection time. Further, the uniformity of hatching was lesser in those eggs which have been kept for longer protection time. While the practical hatching rate increased significantly in silkworm eggs kept for different protection time and the longer refrigeration duration. The hatching rate increased to more than 90% after refrigerating the eggs for 70 days, which was equal to the production demand, also the

  11. 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

  12. 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

  13. 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.

  14. 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.

  15. 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...

  16. Knockout of vlf-1 Gene in Bombyx mori Nucleopolyhedrovirus Using Red Recombination System and Its Influence on Viral Replication%利用Red重组系统敲除家蚕核型多角体病毒的vlf-1基因及对病毒复制的影响

    Institute of Scientific and Technical Information of China (English)

    杜超逸; 于威; 全滟平; 陈健; 聂作明; 吕正兵; 张耀洲

    2013-01-01

    In order to explore the function of BmNPV (Bombyx mori nucleopolyhedrovirus) vlf-1 gene in viral replication, Red recombinant system and BmNPV Bacmid were utilized to have a quick knockout of BmNPV vlf-1 gene in E. coli DHIOBac. Firstly, pKD46 plasmid capable of expressing Red recombinase was isolated from E. coli BW25113 and transformed into E. coli DH10Bac to obtain E. coli DH10Bac (containing pKD46), which could be used for targeting BmNPV gene. Then, for homologous recombination of the target gene, a pair of primers with 60 bp in length was designed, with 40 bp at their 5' end homologous to the left and right arm of vlf-1 gene respectively, and 20 bp at their 3' end homologous to the beginning and ending sequence of cat gene respectively. A linear fragment was amplified by PCR from plasmid pKD3 (containing cat gene) using these primers to obtain the target linear fragment with homologous arms to vlf-1. The linear fragment was transformed into DH10Bac (pKD46) and homologous recombination occurred between the linear fragment and vlf-1 gene in Bacmid DNA with the help of Red recombinase. Two specific PCR primer pairs were used to verify the successful replacement of vlf-1 by cat gene. Then the vlf-1 knockout Bacmid DNA was transfected into BmN cells, and qPCR was employed to analyze the influence of vlf-1 knockout on viral replication. The result indicated that vlf-1 knockout had no influence on initiation of BmNPV DNA replication, but might affect subsequent virus assembly and infection.%利用Red重组系统和家蚕核型多角体病毒(BmNPV) Bacmid在E.coli DH10Bac中快速敲除BmNPV极晚期表达因子基因vlf-1,调查对病毒复制的影响,以探究BmNPV vlf-1基因的功能.首先从E.coli BW25113中提取能表达Red重组酶的质粒pKD46,将其转化到E.coli DH10Bac中,获得可用于BmNPV基因打靶的DH10Bac(含有质粒pKD46)菌株;再为发生基因同源重组设计一对长60 bp的引物,5 '端长40 bp,分别为vlf-1

  17. 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

  18. 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.

  19. 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.

  20. 家蚕吡哆醛激酶基因干扰降低转氨酶基因的转录表达%RNA interference of pyridoxal kinase gene decreases the expression of aminotransferase gene in the silkworm, Bombyx mori

    Institute of Scientific and Technical Information of China (English)

    姚丽丽; 杨欢欢; 张剑韵; 黄龙全

    2015-01-01

    [目的]维生素B6在氨基酸代谢中是多种酶的辅酶,维持氨基酸代谢的正常运行.磷酸吡哆醛(pyridoxal-5'-phosphate,PLP)是维生素B6的主要辅酶形式,吡哆醛激酶(pyridoxal kinase, PLK)是PLP的重要生成酶,本研究试图明确PLK基因与PLP依赖酶之间转录水平的调节关系.[方法]本研究采用RNA干扰(RNA interference,RNAi)方法对家蚕Bombyx mori的PLK基因进行干扰,通过体外合成PLK基因的3个干扰片段(siRNA1,siRNA2和siRNA3),将siRNA从体腔注入5龄第3天的家蚕幼虫体内诱导RNAi.利用荧光定量PCR测定不同干扰片段、不同时间点及不同组织中PLK基因表达量的变化;并测定家蚕体内磷酸丝氨酸转氨酶(phosphoserine aminotransferase,SerB)和天门冬氨酸氨基转移酶(asparate aminotransferase, AST)基因的表达量.[结果]注射干扰片段后48 h干扰效果达到最佳.3个干扰片段干扰效果从高到低依次为siRNA1,siRNA2和siRNA3.RNAi效果最好的是中肠组织,其PLK基因的相对表达量下降了55%.RNA干扰PLK基因后,后部丝腺中SerB和AST基因相对表达量分别下降了90%和29%.[结论]本研究通过RNAi实现了家蚕PLK基因干扰,并进一步证明了家蚕PLK基因和SerB基因及AST基因存在联动调节关系.

  1. 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.

  2. Changes in the transcriptional levels of pyridoxal kinase and pyridoxine-5'-phosphate oxidase post exogenous hormone treatment in the silkworm,Bombyx mori%外源激素处理后家蚕吡哆醛激酶和磷酸吡哆醇氧化酶转录水平的变化

    Institute of Scientific and Technical Information of China (English)

    杨欢欢; 姚丽丽; 张剑韵; 黄龙全

    2015-01-01

    [目的] 研究家蚕Bombyx mori经蜕皮激素(20-hydroxyecdysone,20-E)和保幼激素类似物(juvenile hormone analogue,JHA)处理后引起吡哆醛激酶(pyridoxal kinase,PLK)和磷酸吡哆醇氧化酶(pyridoxine-5'-phosphateoxidase,PNPO)的转录水平变化,为进一步研究激素对蚕体营养代谢等工作奠定基础.[方法]以20-E和JHA分别喂食不同发育时期(5龄第1,3和5天)的家蚕幼虫,以喂食蒸馏水的家蚕为对照,采用实时荧光定量PCR(real-time quantitative PCR)方法在处理后24和48 h对各组幼虫后部丝腺中PLP合成酶PLK和PNPO的转录水平进行分析.[结果]5龄第1天幼虫经20-E处理24和48 h后,PLK和PNPO的转录水平出现上调且与对照的差异达到极显著(P<0.01);5龄第3天幼虫经20-E处理,PLK的转录水平在48 h出现下调且与对照的差异达到显著(P<0.05),PNPO的转录水平在24和48 h均出现上调且与对照的差异达到极显著(P<0.01);5龄第5天幼虫经20-E处理后PLK和PNPO的转录水平无变化.5龄第1天幼虫经JHA处理后PLK和PNPO的转录水平未受到影响;5龄第3天幼虫经JHA处理后,PLK的转录水平在48 h出现显著下调且与对照的差异达到显著(P<0.05),PNPO的转录水平在24和48 h后均出现显著下调且与对照的差异达到极显著(P<0.05);5龄第5天幼虫经JHA处理24和48 h后,PLK和PNPO的转录水平出现下调且与对照的差异达到极显著(P<0.01).[结论]20-E和JHA显著影响家蚕5龄幼虫PLK和PNPO的转录水平,20-E提高5龄前期家蚕PLK和PNPO的转录水平,JHA降低5龄后期它们的转录水平,为深入研究激素对VB6的调控奠定基础.

  3. 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.

  4. 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...

  5. 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...

  6. 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.

  7. Gene expression profile of Bombyx mori hemocyte under the stress of destruxin A.

    Directory of Open Access Journals (Sweden)

    Liang Gong

    Full Text Available Destruxin A (DA is a cyclo-peptidic mycotoxin from the entomopathogenic fungus Metarhizium anisopliae. To uncover potential genes associated with its molecular mechanisms, a digital gene expression (DGE profiling analysis was used to compare differentially expressed genes in the hemocytes of silkworm larvae treated with DA. Ten DGE libraries were constructed, sequenced, and assembled, and the unigenes with least 2.0-fold difference were further analyzed. The numbers of up-regulated genes were 10, 20, 18, 74 and 8, as well as the numbers of down-regulated genes were 0, 1, 8, 13 and 3 at 1, 4, 8, 12 and 24 h post treatment, respectively. Totally, the expression of 132 genes were significantly changed, among them, 1, 3 and 12 genes were continually up-regulated at 4, 3 and 2 different time points, respectively, while 1 gene was either up or down-regulated continually at 2 different time points. Furthermore, 68 genes were assigned to one or multiple gene ontology (GO terms and 89 genes were assigned to specific Kyoto Encyclopedia of Genes and Genomes (KEGG Orthology. In-depth analysis identified that these genes putatively involved in insecticide resistance, cell apoptosis, and innate immune defense. Finally, twenty differentially expressed genes were randomly chosen and validated by quantitative real-time PCR (qRT-PCR. Our studies provide insights into the toxic effect of this microbial insecticide on silkworm's hemocytes, and are helpful to better understanding of the molecular mechanisms of DA as a biological insecticide.

  8. 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

  9. 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.

  10. 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

  11. Ace2, rather than ace1, is the major acetylcholinesterase in the silkworm, Bombyx moil

    Institute of Scientific and Technical Information of China (English)

    Hui-Juan Chen; Zhen Liao; Xiao-Ming Hui; Guo-Qing Li; Fei Li; Zhao-Jun Han

    2009-01-01

    Two acetylcholinesterase (ace) genes have been reported in many insect species. In pests such as Helicoverpa assulta and Plutella xylostellas, acel gene encodes the predominant synaptic enzyme that is the main target of organophosphorus (OP) and carbamate pesticides. It has been reported that pesticide selection has an impact on the ace gene evolution. The domesticated silkworm, Bombyx mori, also has two ace genes. We studied ace gene expression and enzyme activities in silkworm as this has not faced pesticide selection over the past decades. The expression levels of two ace genes, Bm-acel and Bin-ace2, were estimated by quantitative real-time polymerase chain reaction. Bm-ace2 was expressed more highly than Bm-acel in all tested samples of different developmental stages or tissues, suggesting ace2, rather than ace 1, is the major type of acetylcholinesterase (ACHE) in Bombyx mori. This is inconsistent with the aforementioned lepidopterons agricultural pests, partly be due to the widespread use of pesticides that may induce high expression of the acel gene in these pests. Besides high expression in the head, Bm-acel also expresses highly in the silk glands and Bm-ace2 is abundant in the germline, implying both ace genes may have potential non-hydrolytic roles in development. Furthermore, we found that the m_RNA levels of two ace genes and their ratios (ace2/ace1) change day to day in the first and third instars. This challenges the conventional method of estimating enzymatic activity using crude extract as an enzyme solution, as it is a mixture of ACHE1 and ACHE2. An efficient and simple method for separating different ACHEs is necessary for reliable toxicological analyses.

  12. 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.

  13. 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.

  14. 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...

  15. 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

  16. 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...

  17. 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.

  18. 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...

  19. 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)

  20. 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.

  1. 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.

  2. 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

  3. Insect-machine hybrid system for understanding and evaluating sensory-motor control by sex pheromone in Bombyx mori.

    Science.gov (United States)

    Kanzaki, Ryohei; Minegishi, Ryo; Namiki, Shigehiro; Ando, Noriyasu

    2013-11-01

    To elucidate the dynamic information processing in a brain underlying adaptive behavior, it is necessary to understand the behavior and corresponding neural activities. This requires animals which have clear relationships between behavior and corresponding neural activities. Insects are precisely such animals and one of the adaptive behaviors of insects is high-accuracy odor source orientation. The most direct way to know the relationships between neural activity and behavior is by recording neural activities in a brain from freely behaving insects. There is also a method to give stimuli mimicking the natural environment to tethered insects allowing insects to walk or fly at the same position. In addition to these methods an 'insect-machine hybrid system' is proposed, which is another experimental system meeting the conditions necessary for approaching the dynamic processing in the brain of insects for generating adaptive behavior. This insect-machine hybrid system is an experimental system which has a mobile robot as its body. The robot is controlled by the insect through its behavior or the neural activities recorded from the brain. As we can arbitrarily control the motor output of the robot, we can intervene at the relationship between the insect and the environmental conditions.

  4. 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

  5. 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

  6. 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.

  7. 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.

  8. Progress of Research on Bombyx Innate Immunity%家蚕先天性免疫的研究进展

    Institute of Scientific and Technical Information of China (English)

    徐颖

    2012-01-01

    蚕桑生产是我国的传统优势产业,但蚕病却给养蚕业造成很大的损失.由于产业发展的需要,家蚕的免疫防御机制是人们长期关注的热点.家蚕虽然不具有人类高度专一的获得性免疫,但具有对病原微生物感染作出快速有效应答的先天性免疫系统.家蚕受到微生物的感染后,体内会合成抗菌肽,然后抗菌肽被分泌到血淋巴中去消灭病原体.其中,模式识别受体、免疫信号传导途径以及抗菌肽在体液免疫中起着非常重要的作用.%Sericulture production is a traditional industry in China. However,silkworm diseases have caused a great loss in economy. Due to the needs of industrial development, immune defense mechanism of the silkworm is a long-term focus of attention. Bombyx mori, although lacking an a- daptive immune system found in mammals, can resist rapidly and effectively the infection of various microorganisms through multifaceted innate immune response. When silkworms are infected by microorganisms, the body will synthesize antimicrobial peptides which are secreted to destroy pathogens. Pattern recognition receptors, immune signaling pathways and antimicrobial peptide play a very important role in the humoral immune system.

  9. 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.

  10. Transcriptomic Analysis of the Anterior Silk Gland in the Domestic Silkworm (Bombyx mori - Insight into the Mechanism of Silk Formation and Spinning.

    Directory of Open Access Journals (Sweden)

    Huaipu Chang

    Full Text Available Silk proteins are synthesized in the middle and posterior silk glands of silkworms, then transit into the anterior of the silk gland, where the silk fibers are produced, stored and processed. The mechanism of formation and spinning of the silk fibers has not been fully elucidated, and transcriptome analyses specific to the anterior silk gland have not been reported. In the present study, we explored gene expression profiles in five regions of silk gland samples using the RNA-Seq method. As a result, there were 959,979,570 raw reads obtained, of which 583,068,172 reads were mapped to the silkworm genome. A total of 7419 genes were found to be expressed in terms of reads per kilobase of exon model per million mapped reads ≥ 5 in at least one sample. The gene numbers and expression levels of the expressed genes differed between these regions. The differentially expressed genes were analyzed, and 282 genes were detected as up-regulated in the anterior silk gland, compared with the other parts. Functions of these genes were addressed using the gene ontology and Kyoto Encyclopedia of Genes and Genomes databases, and seven key pathways were enriched. It suggested that the ion transportation, energy metabolism, protease inhibitors and cuticle proteins played essential roles in the process of silk formation and spinning in the anterior silk gland. In addition, 210 genes were found differently expressed between males and females, which should help to elucidate the mechanism of the quality difference in silk fibers from male and female silkworms.

  11. 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].

  12. Analysis of bulked segregants to identify molecular markers linked with cocoon weight and cocoon shell weight in the silkworm Bombyx mori L

    Institute of Scientific and Technical Information of China (English)

    SateeshKumar; 徐孟奎; 陈玉银; Ponnuvel,K.M; Datta,R.K

    2002-01-01

    Two silkworm strains viz, B20 A (high cocoon shell ratio) and C. Nichi (low cocoon shell ratio) were sib mated for 10 generations to determine the homozygoeis. Both bulked segregant analysis (BSA) and near isogenic lines (NIL) studies were done to identify the RFLP markers doaely linked to cocoon shell parameters. Three hundred and fifty-two random clones were identified as the low copy number 'sequeiace and used for identification of Restriction Fragment Length Polyrnorphic (RFLP) marker linked to cocoon weight and cocoon shell character. In the bulk aegregant analysia, DNA from the parents (B20 A, C.Nichi), Fl and F2 progeny of high shell ratio (HSR) and low shell ratio (LSR) were screened for hybridization with the random clones. Polymorphic banding pattern achieved through southern hybridization with different probes indicated the probable correlation of polymorphism with high and low cocoon shell character which are possible landmarks in identifying the putative marker(s) for the cocoon shell character. Out of the 100 probes tried with parents, Fl, F2 and their bulks 10 probes were found to be closely linked to cocoon shell characters.

  13. 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)...

  14. 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...

  15. 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.

  16. Studies on Karyotype of Beauveria bassiana,a Pathogenic Fungus to Bombyx mori L.%家蚕病原白僵菌的核型分析

    Institute of Scientific and Technical Information of China (English)

    时连根

    2000-01-01

    用脉冲凝胶电泳中的等高压均匀电场(counter-clamped homogeneous electric field,CHEF)凝胶电泳技术研究了家蚕病原白僵菌(Beauveria bassiana)核型.家蚕病原白僵菌至少有6条染色体,估算其大小在2.5~7.2Mbp之间,3种分离菌株的核型大小为26.5~29.0Mbp.家蚕病原白僵菌核型在分离菌株间存在多型性.

  17. 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.

  18. 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.

  19. 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.

  20. DNA Synthesis during Endomitosis Is Stimulated by Insulin via the PI3K/Akt and TOR Signaling Pathways in the Silk Gland Cells of Bombyx mori

    Directory of Open Access Journals (Sweden)

    Yaofeng Li

    2015-03-01

    Full Text Available Silk gland cells undergo multiple endomitotic cell cycles during silkworm larval ontogeny. Our previous study demonstrated that feeding is required for continued endomitosis in the silk gland cells of silkworm larvae. Furthermore, the insulin signaling pathway is closely related to nutritional signals. To investigate whether the insulin signaling pathway is involved in endomitosis in silk gland cells, in this study, we initially analyzed the effects of bovine insulin on DNA synthesis in endomitotic silk gland cells using 5-bromo-2'-deoxyuridine (BrdU labeling technology, and found that bovine insulin can stimulate DNA synthesis. Insulin signal transduction is mainly mediated via phosphoinositide 3-kinase (PI3K/Akt, the target of rapamycin (TOR and the extracellular signal-regulated kinase (ERK pathways in vertebrates. We ascertained that these three pathways are involved in DNA synthesis in endomitotic silk gland cells using specific inhibitors against each pathway. Moreover, we investigated whether these three pathways are involved in insulin-stimulated DNA synthesis in endomitotic silk gland cells, and found that the PI3K/Akt and TOR pathways, but not the ERK pathway, are involved in this process. These results provide an important theoretical foundation for the further investigations of the mechanism underlying efficient endomitosis in silk gland cells.

  1. Analysis of bulked segregants to identify molecular markers linked with cocoon weight and cocoon shell weight in the silkworm Bombyx mori L

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Two silkworm strains viz, B20 A (high cocoon shell ratio) and C.Nichi (low cocoon shell ratio) were sib mated for 10 generations to determine the homozygosis. Both bulked segregant analysis(BSA) and near isogenic lines (NIL) studies were done to identify the RFLP markers closely linked to cocoon shell parameters. Three hundred and fifty-two random clones were identified as the low copy number sequence and used for identification of Restriction Fragment Length Polymorphic (RFLP) marker linked to cocoon weight and cocoon shell character. In the bulk segregant analysis, DNA from the parents (B20 A, C.Nichi), F1 and F2 progeny of high shell ratio (HSR) and low shell ratio (LSR) were screened for hybridization with the random clones. Polymorphic banding pattern achieved through southern hybridization with different probes indicated the probable correlation of polymorphism with high and low cocoon shell character which are possible landmarks in identifying the putative marker(s) for the cocoon shell character. Out of the 100 probes tried with parents, F1, F2 and their bulks, 10 probes were found to be closely linked to cocoon shell characters.

  2. 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

  3. A Rapid Isolation Method of Silkworm (Bombyx mori) Genomic DNA%家蚕蛹体基因组DNA的快速制备方法

    Institute of Scientific and Technical Information of China (English)

    赵巧玲; 张志芳; 何家禄

    2000-01-01

    @@ 提取家蚕组织DNA的方法已有不少报道,如夏庆友等[1]参照文献[2]改进的方法,其基本步骤是将家蚕组织在液氮中手工研磨或用玻璃匀浆器匀浆,加提取缓冲液和Proteinase K,在50~65℃水浴恒温振荡3~6h,然后用酚、氯仿抽提数次,无水乙醇沉淀,75%乙醇洗涤后溶于适当体积的0.1×TE缓冲液,加RNA酶消解RNA,保存备用.该方法适于提取丝腺、卵的基因组DNA.然而,家蚕蛹体由于脂肪、色素较多,且整体组织中含有大量的DNA酶,按上述方法提取DNA纯度不高,并容易降解.我们分析认为可能是提取液中的EDTA浓度太低,不足以抑制整体组织中的DNA酶.刘春宇等[3]提取家蚕蛹体DNA则是预先在1×SSC溶液中匀浆、离心、漂洗,然后将沉淀进行抽提.该方法有效地去除部分蛋白质、脂肪、色素等杂质.上述方法都使用了Proteinase K,所以试验费用较昂贵,整个提取过程花费时间也较长.为此,我们设计了一套经济、快速地制备家蚕蛹体基因组DNA的方法,经试验此法也适用于各不同发育阶段的家蚕组织.

  4. 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

  5. Complete resequencing of 40 genomes reveals domestication events and genes in silkworm (Bombyx)

    DEFF Research Database (Denmark)

    Xia, Qingyou; Guo, Yiran; Zhang, Ze;

    2009-01-01

    A single-base pair resolution silkworm genetic variation map was constructed from 40 domesticated and wild silkworms, each sequenced to approximately threefold coverage, representing 99.88% of the genome. We identified ~16 million single-nucleotide polymorphisms, many indels, and structural...... variations. We find that the domesticated silkworms are clearly genetically differentiated from the wild ones, but they have maintained large levels of genetic variability, suggesting a short domestication event involving a large number of individuals. We also identified signals of selection at 354 candidate...... genes that may have been important during domestication, some of which have enriched expression in the silk gland, midgut, and testis. These data add to our understanding of the domestication processes and may have applications in devising pest control strategies and advancing the use of silkworms...

  6. 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

  7. 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.

  8. EST Table: FS927712 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available aa ref|NP_001037416.1| achaete-scute-like protein [Bombyx mori] gb|ABC84346.1| achaete-scute-like protein [...Bombyx mori] gb|ABC84347.1| achaete-scute-like protein [Bombyx mori] 10/09/13 35

  9. 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...

  10. 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

  11. Molecular identification of the first SIFamide receptor

    DEFF Research Database (Denmark)

    Jørgensen, Lars M; Hauser, Frank; Cazzamali, Giuseppe;

    2006-01-01

    . Database searches revealed SIFamide receptor orthologues in the genomes from the malaria mosquito Anopheles gambiae, the silkworm Bombyx mori, the red flour beetle Tribolium castaneum, and the honey bee Apis mellifera. An alignment of the five insect SIFamide or SIFamide-like receptors showed, again...

  12. An improved Mori-Tanaka method for nonlinear interface debonding in composite solid propellant%改进的Mori-Tanaka法在复合推进剂非线性界面脱粘中的应用

    Institute of Scientific and Technical Information of China (English)

    刘承武; 阳建红; 陈飞

    2011-01-01

    Aiming at the problems concerning the composite solid propellant with large deformation in the matrix and interface debonding around the filled particles, an improved Mori-Tanaka method was developed. At the same time, in order to verify its effectiveness, a simulation method for the interface debonding of the composite solid propellant consists of random filled particles was also put forward. Finally, a computational case of one kind of composite solid propellant was investigated. It is demonstrated that the resuits from these two methods are close and the improved Mori-Tanaka method is effective; the computational time can be reduced dramatically and the computational effectiveness can be increased greatly by using the improved Mori-Tanaka method.%针对复合固体推进剂基体的大变形和界面的脱粘问题,对Mori-Tanaka法进行了改进.同时,为验证该方法的有效性,针对推进剂颗粒随机填充的特点,提出了一种含非线性界面脱粘的数值方法.最后以某推进剂配方为算例,对2种方法的计算结果进行了比较.结果表明,2种方法结果接近,从而验证了改进的Mori-Tanaka法的有效性,且改进的Mori-Tanaka法计算量小,极大地提高了计算效率.

  13. Optimization of enzyme assisted extraction of Fructus Mori polysaccharides and its activities on antioxidant and alcohol dehydrogenase.

    Science.gov (United States)

    Deng, Qingfang; Zhou, Xin; Chen, Huaguo

    2014-10-13

    In the present study, enzyme assisted extraction of Fructus Mori polysaccharides (FMPS) from F. mori using four kinds of enzymes and three compound enzymes were examined. Research found that glucose oxidase offered a better performance in enhancement of the extraction yields of FMPS, antioxidant and activate alcohol dehydrogenase activities. The glucose oxidase assisted extraction process was further optimized by using response surface method (RSM) to obtain maximum yield of crude FMPS. The results showed that optimized extraction conditions were ratio of enzyme amount 0.40%, enzyme treated time 38 min, treated temperature 58 °C and liquid-solid radio 11.0. Under these conditions, the mean experimental value of extraction yield (16.16 ± 0.14%) corresponded well with the predicted values and increased 160% than none enzyme treated ones. Pharmacological verification tests showed that F. mori crude polysaccharides had good antioxidant and activate alcohol dehydrogenase activities in vitro.

  14. 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.

  15. Memory-enhancing effect of Mori Fructus via induction of nerve growth factor.

    Science.gov (United States)

    Kim, Hyo Geun; Oh, Myung Sook

    2013-07-14

    Fruits rich in phytochemicals have been shown to improve memory by protecting or enhancing neuronal functions mediated by neurotrophic factors, such as nerve growth factor (NGF), in the hippocampus. Mori Fructus (Morus alba L., Moraceae), also called mulberry, is used as a food, dietary supplement and an anti-ageing agent in traditional Oriental medicine. It is also known to contain abundant flavonoid compounds and to exhibit various pharmacological effects. The present study was performed to evaluate the memory-enhancing effect of Mori Fructus extract (ME) in mice, with a focus on NGF regulation. ME (20, 100 and 500 mg/kg per d for 7 d, per os) dose-dependently promoted NGF release in the mouse hippocampus, leading to phosphorylation of extracellular signal-regulated kinases and cyclic AMP response element-binding protein. ME significantly increased pre- and post-synapse formation, acetylcholine synthesisation, neuronal cell differentiation, neurite outgrowth and neuronal cell proliferation in the mouse hippocampus. Furthermore, ME significantly increased latency time in the passive avoidance task (Pimprovements in learning and memory. Taken together, these data suggest that ME exhibits a memory-enhancing effect via up-regulation of NGF.

  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: 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- ...

  18. 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 ...

  19. 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 ...

  20. 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 ...

  1. 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 ...

  2. 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 ...

  3. 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 ...

  4. 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 ...

  5. 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- ...

  6. EST Table: DC536003 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ombyx mori] dbj|BAE53372.1| antibacterial peptide [Bombyx mori] dbj|BAF51564.1| gloverin2 [Bombyx mori] 10/09/01 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS917189 MFB- ...

  7. 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 ...

  8. 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- ...

  9. 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 ...

  10. 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- ...

  11. 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 ...

  12. 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 ...

  13. EST Table: CK501907 [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 ...

  14. 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 ...

  15. EST Table: DC537189 [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- ...

  16. 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 ...

  17. 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 ...

  18. 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 ...

  19. 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 ...

  20. 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- ...

  1. 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 ...

  2. EST Table: CK504037 [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: DC538487 [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- ...

  4. 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 ...

  5. 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- ...

  6. 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 ...

  7. 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 ...

  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: 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 ...

  10. 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 ...

  11. 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 ...

  12. 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 ...

  13. 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 ...

  14. 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 ...

  15. EST Table: CK505508 [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 ...

  16. 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 ...

  17. 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 ...

  18. 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 ...

  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. 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 ...

  1. 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 ...

  2. EST Table: DC538561 [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- ...

  3. 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 ...

  4. 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 ...

  5. 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 ...

  6. 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 ...

  7. 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 ...

  8. 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 ...

  9. 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 ...

  10. 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 ...

  11. 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 ...

  12. 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 ...

  13. EST Table: CK507076 [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 ...

  14. 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 ...

  15. EST Table: CK501867 [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 ...

  16. 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 ...

  17. 家蚕胚胎细胞系BmE-SWU1的建立及其生物学特性%Establishment and Characterization of Embryo Cell Line of Bombyx mori -SWU1

    Institute of Scientific and Technical Information of China (English)

    潘敏慧; 肖仕全; 洪锡钧; 鲁成

    2005-01-01

    对家蚕反转期胚胎组织进行一年多的原代培养,分离筛选出了高繁殖细胞群体,建立了BmE-SWU1细胞系.该细胞系以梭形和近圆形细胞为主,杂以少量多突起和巨型细胞.细胞系的倍增时间在28℃时为57.6 h,染色体呈短杆状或颗粒状,染色体数目异倍化,具有典型鳞翅目昆虫细胞系的染色体特征.BmE-SWU1细胞系对家蚕核型多角体病毒(BmNPV)高度敏感,半数组织培养感染剂量(TCID50)为2.92415×10-7.

  18. 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.

  19. Studies on the Regeneration and Reversion of Protoplasts From Beauveria bassiana,a Pathogenic Fungus to Bombyx mori L.%家蚕病原白僵菌原生质体的再生回复研究

    Institute of Scientific and Technical Information of China (English)

    时连根; 柯林田; 费建明

    1999-01-01

    研究了家蚕病原白僵菌(Beauveria bassiana)原生质体的性状、再生回复形式与条件.家蚕病原白僵菌原生质体的无核率为26.5%,有核率为73.5%,其中单核率为53.5%.原生质体再生回复的形式可观察到3种,在位相差显微镜下不能判别原生质体有无再生回复能力.综合考虑原生质体再生频率和菌落生长发育速度,家蚕病原白僵菌原生质体再生回复用培养基的渗透压稳定剂以0.7mol/L葡萄糖较为适当.

  20. Distribution,Movement and Metabolism of Folate Compounds in the Silkworm,Bombyx mori%家蚕体内叶酸的存在形态和转换代谢

    Institute of Scientific and Technical Information of China (English)

    张剑韵; 黄龙全

    2003-01-01

    采用高效液相色谱(high-performance liquid chromatography,HPLC)技术,探讨了家蚕体内叶酸化合物的存在形态和转换代谢途径.用添加蝶酰谷氨酸(pteroylglutamic acid,PteGlu)的人工合成饲料饲育5龄幼虫,其体内叶酸的主要存在型,血淋巴为PteGlu;血淋巴以外的组织为四氢叶酸(H4PteGlu).脂肪体是H4PteGlu的主要储存场所.叶酸转换代谢酶的检索分析结果也证实家蚕缺乏哺乳动物体内的5,10-亚甲四氢叶酸(5,10-CH2-H4PteGlu)→5-甲基四氢叶酸(5-CH3-H4PteGlu)→H4PteGlu转换途径.家蚕体内叶酸代谢方式简单,推测主要和核酸合成系、丝氨酸/甘氨酸转换系相联系.

  1. Transformation of Neomycin Resistance Gene (neoR) into Silkworm (Bombyx mori.L.)%新霉素抗性基因在家蚕中的插入和表达

    Institute of Scientific and Technical Information of China (English)

    陈秀; 赵昀; 张峰; 彭卫平; 冯晓黎; 黄君霆; 陆长德

    1999-01-01

    构建含新霉素抗性基因(neomycin resistance gene, neoR)的重组质粒pFN, 经HindIII酶切后, 用基因枪将DNA片段导入家蚕早期受精卵中(G0代). 孵化的G1、G2代蚁蚕均经含新霉素的人工饲料添食24 h后, 筛选出新霉素抗性的个体(能正常生长发育的)改为桑叶饲养. 于G2代的5龄第二天从后部丝腺抽提总DNA, 再以neoR的cDNA为探针进行Southern杂交检测. 结果表明neoR基因已转入家蚕DNA中, 获得了含neoR的转基因蚕.

  2. 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.

  3. Gene cloning and expression characteristics of vacuolar-type ATPase subunit B in Bombyx mori%家蚕V型ATP酶B亚基的克隆及表达特征

    Institute of Scientific and Technical Information of China (English)

    陈慧芳; 王鑫; 谢康; 李懿; 赵萍

    2016-01-01

    V型ATP酶(Vacuolar-type ATPase)是一种定位于细胞膜和细胞器膜上的氢离子转运酶.它利用ATP水解的能量将氢离子转运到液泡、囊泡或者胞外,从而维持细胞内正常的酸碱环境.V型ATP酶B亚基(V-ATPase B)作为ATP的催化位点,也有着非常重要的作用.为了探讨家蚕V-ATPase B(BmV-ATPase B)的功能,首先从家蚕五龄幼虫的中肠cDNA中克隆了Bm V-A TPase B基因并构建原核表达载体进行原核表达,获得了重组蛋白,经质谱鉴定正确后,通过镍柱亲和层析的方法纯化了该蛋白并制备了多克隆抗体;最后分析了该蛋白在家蚕丝腺中的表达特征并利用免疫荧光对其在丝腺中的表达位置进行了定位.结果显示Bm V-A TPase B基因序列全长1 473 bp,预测蛋白分子量55 kDa,预测等电点5.3.通过Western blotting对家蚕5龄第3天和上蔟第1天幼虫丝腺的不同区段进行BmV-ATPase B蛋白的表达特征分析,发现在两个时期该蛋白均在前部丝腺高量表达,而在中部丝腺和后部丝腺表达量相对较低.进一步对两个时期丝腺的不同区段进行免疫荧光定位,发现该蛋白在两个时期的前部丝腺、中部丝腺和后部丝腺均定位于细胞层.利用激光共聚焦显微镜对该蛋白进行进一步的定位,发现该蛋白主要在丝腺的细胞膜表达.研究结果明确了该蛋白在丝腺中的表达模式,为深入研究该蛋白在蚕丝纤维形成中的作用奠定了基础.

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. Approximate but accurate quantum dynamics from the Mori formalism: II. Equilibrium correlation functions

    CERN Document Server

    Montoya-Castillo, Andrés

    2016-01-01

    The ability to efficiently and accurately calculate equilibrium time correlation functions of many-body condensed phase quantum systems is one of the outstanding problems in theoretical chemistry. The Nakajima-Zwanzig-Mori formalism coupled to the self-consistent solution of the memory kernel has recently proven to be highly successful for the computation of nonequilibrium dynamical averages. Here, we extend this formalism to treat symmetrized equilibrium time correlation functions for the spin-boson model. Following the first paper in this series [A. Montoya-Castillo and D. R. Reichman, J. Chem. Phys. $\\bf{144}$, 184104 (2016)], we use a Dyson-type expansion of the projected propagator to obtain a self-consistent solution for the memory kernel that requires only the calculation of normally evolved auxiliary kernels. We employ the approximate mean-field Ehrenfest method to demonstrate the feasibility of this approach. Via comparison with numerically exact results for the correlation function $\\mathcal{C}_{zz}...

  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: 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 ...

  11. Identification of a Beauveria bassiana Strain with High Pathogenicity to Bombyx mori and Its Application in Bombyx Batryticatus Production%一株感染家蚕的高致病性白僵菌的鉴定及在僵蚕生产中的应用

    Institute of Scientific and Technical Information of China (English)

    邢东旭; 李丽; 廖森泰; 肖阳; 李庆荣; 叶明强; 杨琼

    2014-01-01

    僵蚕是传统中药材,筛选对家蚕具有高致病性的球孢白僵菌用于提高规模化生产僵蚕的产量和品质.从蚕区采集的僵蚕虫体分离纯化到一株高致病力的白僵菌,通过形态学鉴定和rRNA ITS序列系统发育分析,确定该菌株为球孢白僵菌(Beauveria bassiana),命名为Beauveria bassiana JC-1.优化该菌株应用于僵蚕生产中的接种菌液浓度为1×106~5×106 mL-1,用此浓度的菌液喷洒接种5龄起蚕后5~6d,幼虫大量死亡,僵化率达90%以上,万头蚕可生产僵蚕约5 kg.利用该菌株感染家蚕收获所得僵蚕的各项质量指标均符合《中国药典》中的有关标准,且僵蚕的总灰分和酸不溶性灰分含量较市售僵蚕更低.初步认为该菌株在僵蚕的标准化、规模化生产中具有应用价值.

  12. EST Table: FS775191 [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: FS793531 [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 Am...-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Sclerosis pdb|3...L9E|C Chain C, Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implication... Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral

  14. 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

  15. EST Table: FS741802 [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: FS815906 [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: 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

  18. EST Table: FS734336 [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: FS864010 [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 Am...-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Sclerosis pdb|3...L9E|C Chain C, Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implication... Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral

  20. EST Table: CK521128 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available stal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyot...ficient 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... Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Scl

  1. EST Table: FS845116 [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

  2. 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

  3. 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

  4. EST Table: FS911257 [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

  5. EST Table: FS735529 [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

  6. EST Table: FS765577 [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

  7. EST Table: FS725152 [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

  8. EST Table: FS758475 [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

  9. EST Table: FS771424 [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

  10. EST Table: FS782580 [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 Am...-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Sclerosis pdb|3...L9E|C Chain C, Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implication... Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral

  11. 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

  12. EST Table: FS761730 [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: DC545970 [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

  14. EST Table: FS761100 [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: FS785769 [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 Am...-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Sclerosis pdb|3...L9E|C Chain C, Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implication... Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral

  16. EST Table: FS779295 [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 Am...-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Sclerosis pdb|3...L9E|C Chain C, Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implication... Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral

  17. EST Table: FS765937 [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

  18. EST Table: FS780105 [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 Am...-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Sclerosis pdb|3...L9E|C Chain C, Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implication... Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral

  19. EST Table: FY753052 [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: 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

  1. EST Table: FS743948 [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

  2. EST Table: BP183060 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available uctures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic L...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 Implications I...d Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Sclerosis g

  3. EST Table: FS726254 [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

  4. EST Table: FS797353 [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

  5. EST Table: FS876627 [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 Am...-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Sclerosis pdb|3...L9E|C Chain C, Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implication... Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral

  6. EST Table: FS731913 [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

  7. EST Table: FS781403 [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

  8. EST Table: FS762362 [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

  9. EST Table: FS896634 [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

  10. EST Table: FY015390 [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

  11. EST Table: BW998429 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ctures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic La...uZNSOD 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 Implications In... Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Sclerosis gb

  12. EST Table: FS774242 [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: FS785259 [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

  14. EST Table: BJ983515 [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: 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

  16. EST Table: FS938551 [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: FS768794 [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

  18. EST Table: FS882372 [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: FS762383 [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: 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

  1. 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

  2. EST Table: BJ983684 [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

  3. EST Table: FS778676 [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

  4. EST Table: FS793035 [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

  5. 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

  6. EST Table: BB990147 [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 Am...-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Sclerosis pdb|3...L9E|C Chain C, Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implication... Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral

  7. EST Table: FS730046 [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

  8. EST Table: FS776959 [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 Am...-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Sclerosis pdb|3...L9E|C Chain C, Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implication... Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral

  9. EST Table: FS868002 [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

  10. EST Table: FS876044 [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: FS756226 [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: FS787632 [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: FS863034 [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 Am...-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral Sclerosis pdb|3...L9E|C Chain C, Crystal Structures Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implication... Of Holo And Cu-Deficient CuZNSOD FROM The Silkworm Bombyx Mori And The Implications In Amyotrophic Lateral

  14. EST Table: FS739224 [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: FS884641 [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: FS729718 [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: FS868769 [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

  18. EST Table: FS762270 [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: FS724989 [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: FS744435 [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