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Sample records for sorghum cloning expression

  1. Map-based cloning and expression analysis of BMR-6 in sorghum.

    Science.gov (United States)

    Li, Jieqin; Wang, Lihua; Zhang, Qiuwen; Liu, Yanlong

    2015-09-01

    Brown midrib mutants in sorghum are associated with reduced lignin content and increased cell wall digestibility. In this study, we characterized a bmr-6 sorghum mutant, which shows reddish pigment in the midrib and stem after the fifth-leaf stage. Compared to wild type, Kalson lignin content of bmr-6 is decreased significantly. We used histological analysis to determine that the mutant exhibited a modified pattern of lignin staining and found an increased polysaccharide content. We cloned BMR-6 gene, a gene encoded a cinnamyl alcohol dehydrogenase (CAD), using a map-based cloning approach. Genetic complementation confirmed that CAD is responsible for the BMR-6 phenotype. BMR-6 gene was expressed in all tested sorghum tissues, with the highest being in midrib and stem. Transient expression assays in Nicotiana benthamiana leaves demonstrated cytomplasmic localization of BMR-6. We found that the expression level of bmr-6 was significantly decreased in the mutant but expression of SbCAD3 and SbCAD5 were significantly increased. Our results indicate that BMR-6 not only affects the distribution of lignin but also the biosynthesis of lignin in sorghum.

  2. Map-based cloning and expression analysis of BMR-6 in sorghum

    Indian Academy of Sciences (India)

    CAD), using a map-based cloning approach. Genetic complementation confirmed that CAD is responsible for the BMR-6 phenotype. BMR-6 gene was expressed in all tested sorghum tissues, with the highest being in midrib and stem. Transient ...

  3. Diurnal oscillation of SBE expression in sorghum endosperm

    Energy Technology Data Exchange (ETDEWEB)

    Sun, Chuanxin; Mutisya, J.; Rosenquist, S.; Baguma, Y.; Jansson, C.

    2009-01-15

    Spatial and temporal expression patterns of the sorghum SBEI, SBEIIA and SBEIIB genes, encoding, respectively, starch branching enzyme (SBE) I, IIA and IIB, in the developing endosperm of sorghum (Sorghum bicolor) were studied. Full-length genomic and cDNA clones for sorghum was cloned and the SBEIIA cDNA was used together with gene-specific probes for sorghum SBEIIB and SBEI. In contrast to sorghum SBEIIB, which was expressed primarily in endosperm and embryo, SBEIIA was expressed also in vegetative tissues. All three genes shared a similar temporal expression profile during endosperm development, with a maximum activity at 15-24 days after pollination. This is different from barley and maize where SBEI gene activity showed a significantly later onset compared to that of SBEIIA and SBEIIB. Expression of the three SBE genes in the sorghum endosperm exhibited a diurnal rhythm during a 24-h cycle.

  4. Cloning and expression analysis of cinnamoyl-CoA reductase (CCR) genes in sorghum.

    Science.gov (United States)

    Li, Jieqin; Fan, Feifei; Wang, Lihua; Zhan, Qiuwen; Wu, Peijin; Du, Junli; Yang, Xiaocui; Liu, Yanlong

    2016-01-01

    Cinnamoyl-CoA reductase (CCR) is the first enzyme in the monolignol-specific branch of the lignin biosynthetic pathway. In this research, three sorghum CCR genes including SbCCR1, SbCCR2-1 and SbCCR2-2 were cloned and characterized. Analyses of the structure and phylogeny of the three CCR genes showed evolutionary conservation of the functional domains and divergence of function. Transient expression assays in Nicotiana benthamiana leaves demonstrated that the three CCR proteins were localized in the cytoplasm. The expression analysis showed that the three CCR genes were induced by drought. But in 48 h, the expression levels of SbCCR1 and SbCCR2-2 did not differ between CK and the drought treatment; while the expression level of SbCCR2-1 in the drought treatment was higher than in CK. The expression of the SbCCR1 and SbCCR2-1 genes was not induced by sorghum aphid [Melanaphis sacchari (Zehntner)] attack, but SbCCR2-2 was significantly induced by sorghum aphid attack. It is suggested that SbCCR2-2 is involved in the process of pest defense. Absolute quantitative real-time PCR revealed that the three CCR genes were mainly expressed in lignin deposition organs. The gene copy number of SbCCR1 was significantly higher than those of SbCCR2-1 and SbCCR2-2 in the tested tissues, especially in stem. The results provide new insight into the functions of the three CCR genes in sorghum.

  5. Cloning and expression of a sorghum gene with homology to maize vp1. Its potential involvement in pre-harvest sprouting resistance.

    Science.gov (United States)

    Carrari, F; Perez-Flore, L; Lijavetzky, D; Enciso, S; Sanchez, R; Benech-Arnold, R; Iusem, N

    2001-04-01

    Pre-harvest sprouting (PHS) in sorghum is related to the lack of a normal dormancy level during seed development and maturation. Based on previous evidence that seed dormancy in maize is controlled by the vp1 gene, we used a PCR-based approach to isolate two Sorghum bicolor genomic and cDNA clones from two genotypes exhibiting different PHS behaviour and sensitivity to abscisic acid (ABA). The two 699 amino acid predicted protein sequences differ in two residues at positions 341 (Gly or Cys within the repression domain) and 448 (Pro or Ser) and show over 80, 70 and 60% homology to maize, rice and oat VP1 proteins respectively. Expression analysis of the sorghum vp1 gene in the two lines shows a slightly higher level of vp1 mRNA in the embryos susceptible to PHS than in those resistant to PHS during embryogenesis. However, timing of expression was different between these genotypes during this developmental process. Whereas for the former the main peak of expression was observed at 20 days after pollination (DAP), the peak in the latter was found at later developmental stages when seed maturation was almost complete. Under favourable germination conditions and in the presence of fluridone (an inhibitor of ABA biosynthesis), sorghum vp1 mRNA showed to be consistently correlated with sensitivity to ABA but not with ABA content and dormancy.

  6. A Sorghum bicolor expression atlas reveals dynamic genotype-specific expression profiles for vegetative tissues of grain, sweet and bioenergy sorghums

    Energy Technology Data Exchange (ETDEWEB)

    Shakoor, N; Nair, R; Crasta, O; Morris, G; Feltus, A; Kresovich, S

    2014-01-23

    Background: Effective improvement in sorghum crop development necessitates a genomics-based approach to identify functional genes and QTLs. Sequenced in 2009, a comprehensive annotation of the sorghum genome and the development of functional genomics resources is key to enable the discovery and deployment of regulatory and metabolic genes and gene networks for crop improvement. Results: This study utilizes the first commercially available whole-transcriptome sorghum microarray (Sorgh-WTa520972F) to identify tissue and genotype-specific expression patterns for all identified Sorghum bicolor exons and UTRs. The genechip contains 1,026,373 probes covering 149,182 exons (27,577 genes) across the Sorghum bicolor nuclear, chloroplast, and mitochondrial genomes. Specific probesets were also included for putative non-coding RNAs that may play a role in gene regulation (e. g., microRNAs), and confirmed functional small RNAs in related species (maize and sugarcane) were also included in our array design. We generated expression data for 78 samples with a combination of four different tissue types (shoot, root, leaf and stem), two dissected stem tissues (pith and rind) and six diverse genotypes, which included 6 public sorghum lines (R159, Atlas, Fremont, PI152611, AR2400 and PI455230) representing grain, sweet, forage, and high biomass ideotypes. Conclusions: Here we present a summary of the microarray dataset, including analysis of tissue-specific gene expression profiles and associated expression profiles of relevant metabolic pathways. With an aim to enable identification and functional characterization of genes in sorghum, this expression atlas presents a new and valuable resource to the research community.

  7. MADS box genes expressed in developing inflorescences of rice and sorghum

    NARCIS (Netherlands)

    Greco, R.; Stagi, L.; Colombo, L.; Angenent, G.C.; Sari-Gorla, M.; Pé, M.E.

    1997-01-01

    With the aim of elucidating the complex genetic system controlling flower morphogenesis in cereals, we have characterized two rice and two sorghum MADS box genes isolated from cDNA libraries made from developing inflorescences. The rice clones OsMADS24 and OsMADS45, which share high homology with

  8. Identification of differentially expressed genes in sorghum (Sorghum bicolor) brown midrib mutants

    Science.gov (United States)

    Sorghum (Sorghum bicolor L.), with a high biomass yield and excellent tolerance to drought and low nutrition, has been recommended as one of the most competitive bioenergy crops. Brown midrib (bmr) mutant sorghum with reduced lignin content showed a high potential for the improvement of bioethanol ...

  9. Discovery of cis-elements between sorghum and rice using co-expression and evolutionary conservation

    Directory of Open Access Journals (Sweden)

    Haberer Georg

    2009-06-01

    Full Text Available Abstract Background The spatiotemporal regulation of gene expression largely depends on the presence and absence of cis-regulatory sites in the promoter. In the economically highly important grass family, our knowledge of transcription factor binding sites and transcriptional networks is still very limited. With the completion of the sorghum genome and the available rice genome sequence, comparative promoter analyses now allow genome-scale detection of conserved cis-elements. Results In this study, we identified thousands of phylogenetic footprints conserved between orthologous rice and sorghum upstream regions that are supported by co-expression information derived from three different rice expression data sets. In a complementary approach, cis-motifs were discovered by their highly conserved co-occurrence in syntenic promoter pairs. Sequence conservation and matches to known plant motifs support our findings. Expression similarities of gene pairs positively correlate with the number of motifs that are shared by gene pairs and corroborate the importance of similar promoter architectures for concerted regulation. This strongly suggests that these motifs function in the regulation of transcript levels in rice and, presumably also in sorghum. Conclusion Our work provides the first large-scale collection of cis-elements for rice and sorghum and can serve as a paradigm for cis-element analysis through comparative genomics in grasses in general.

  10. Diversity of nifH gene pools in the rhizosphere of two cultivars of sorghum (Sorghum bicolor) treated with contrasting levels of nitrogen fertilizer.

    Science.gov (United States)

    Coelho, Marcia Reed Rodrigues; de Vos, Marjon; Carneiro, Newton Portilho; Marriel, Ivanildo Evódio; Paiva, Edilson; Seldin, Lucy

    2008-02-01

    The diversity of nitrogen-fixing bacteria was assessed in the rhizospheres of two cultivars of sorghum (IS 5322-C and IPA 1011) sown in Cerrado soil amended with two levels of nitrogen fertilizer (12 and 120 kg ha(-1)). The nifH gene was amplified directly from DNA extracted from the rhizospheres, and the PCR products cloned and sequenced. Four clone libraries were generated from the nifH fragments and 245 sequences were obtained. Most of the clones (57%) were closely related to nifH genes of uncultured bacteria. NifH clones affiliated with Azohydromonas spp., Ideonella sp., Rhizobium etli and Bradyrhizobium sp. were found in all libraries. Sequences affiliated with Delftia tsuruhatensis were found in the rhizosphere of both cultivars sown with high levels of nitrogen, while clones affiliated with Methylocystis sp. were detected only in plants sown under low levels of nitrogen. Moreover, clones affiliated with Paenibacillus durus could be found in libraries from the cultivar IS 5322-C sown either in high or low amounts of fertilizer. This study showed that the amount of nitrogen used for fertilization is the overriding determinative factor that influenced the nitrogen-fixing community structures in sorghum rhizospheres cultivated in Cerrado soil.

  11. Expression Pattern of the Alpha-Kafirin Promoter Coupled with a Signal Peptide from Sorghum bicolor L. Moench

    Directory of Open Access Journals (Sweden)

    Norazlina Ahmad

    2012-01-01

    Full Text Available Regulatory sequences with endosperm specificity are essential for foreign gene expression in the desired tissue for both grain quality improvement and molecular pharming. In this study, promoters of seed storage α-kafirin genes coupled with signal sequence (ss were isolated from Sorghum bicolor L. Moench genomic DNA by PCR. The α-kafirin promoter (α-kaf contains endosperm specificity-determining motifs, prolamin-box, the O2-box 1, CATC, and TATA boxes required for α-kafirin gene expression in sorghum seeds. The constructs pMB-Ubi-gfp and pMB-kaf-gfp were microprojectile bombarded into various sorghum and sweet corn explants. GFP expression was detected on all explants using the Ubi promoter but only in seeds for the α-kaf promoter. This shows that the α-kaf promoter isolated was functional and demonstrated seed-specific GFP expression. The constructs pMB-Ubi-ss-gfp and pMB-kaf-ss-gfp were also bombarded into the same explants. Detection of GFP expression showed that the signal peptide (SP::GFP fusion can assemble and fold properly, preserving the fluorescent properties of GFP.

  12. Development, applications and distribution of DNA markers for genetic information for sorghum and maize improvement

    International Nuclear Information System (INIS)

    Lee, M.

    2001-01-01

    This final report summarizes the progress made towards the enhancement and distribution of genetic resources (e.g. genetic stocks, seed and DNA clones) used for basic and applied aspects of the genetic improvement of maize and sorghum. The genetic maps of maize and sorghum were improved through comparative mapping of RFLP loci detected by 124 maize cDNA clones and through the development of a new mapping population of maize. Comparative mapping between maize and sorghum and maize and rice, using the set of 124 maize cDNA clones (and other clones) in each study, substantiated previous observations of extensive conservation of locus order but it also provided strong evidence of numerous large-scale chromosomal rearrangements. The new mapping population for maize (intermated B73xMo17, 'IBM') was created by random intermating during the first segregating generation. Intermating for four generations prior to the derivation of recombinant inbred lines (RILs) increased the frequency of recombinants at many regions of the maize genome and provided better genetic resolution of locus order. Expansion of the maize genetic map was not uniform along the length of a linkage group and was less than the theoretical expectation. The 350 IBM RILs were genotyped at 512 loci detected by DNA clones, including 76 of the 124 supported by this contract. The production of the sorghum mapping population of RILs from the cross CK60xPI229828 has been delayed by weather conditions that were not conducive to plant growth and seed development. Seed of the IBM RILs have been distributed (approximately 5000 RILs in total) to 16 research organizations in the public and private sector. The DNA clones have been distributed (1,206 in total) to nine research labs. Further distribution of the seed and clones will be managed by curators at stock centers in the public domain. (author)

  13. Circadian oscillation of starch branching enzyme gene expression in the sorghum endosperm

    Energy Technology Data Exchange (ETDEWEB)

    Mutisya, J.; Sun, C.; Jansson, C.

    2009-08-31

    Expression of the three SBE genes, encoding starch branching enzymes, in the sorghum endosperm exhibited a diurnal rhythm during a 24-h cycle. Remarkably, the oscillation in SBE expression was maintained in cultured spikes after a 48-h dark treatment, also when fed a continuous solution of sucrose or abscisic acid. Our findings suggest that the rhythmicity in SBE expression in the endosperm is independent of cues from the photosynthetic source and that the oscillator resides within the endosperm itself.

  14. Genetic dissection of bioenerrgy traits in sorghum

    Energy Technology Data Exchange (ETDEWEB)

    Vermerris, Wilfred; Kresovich, Stephen; Murray, Seth; Pedersen, Jeffery; Rooney, William; Sattler, Scott.

    2012-06-15

    Specific Objectives: 1. To identify the gene(s) underlying a major QTL for stem sugar concentration located on chromosome 3. 2. To identify QTL for stem juice volume and stalk sugar concentration and to identify the underlying genes. 3. To classify 60 novel sorghum bmr mutants from the USDA TILLING population in allelic groups based on cell wall chemistry and allelism tests. 4. To select representative bmr mutants from each allelic group and selected NIR spectral mutants for their potential value as feedstock for ethanol production. 5. To clone and characterize those Bmr genes that represent loci other than Bmr12 and Bmr6 using a mapping and a candidate gene approach. Objective 1 The experiments for this objective are largely complete and the data have been analyzed. Data interpretation and follow-up experiments are still in progress. A manuscript is in preparation (Vermerris et al.; see publication list for full details). The main results are: 1) 16 cDNA libraries were prepared and sequenced at Cornell University. The libraries represent internode tissue and flag leaf tissue at booting, internode tissue and peduncle at soft-dough stage, from two plants per sampling time with the Rio allele for the QTL on chromosome 3, and two plants with the BTx623 allele on chromosome 3 (4 tissues x 2 genotypes x 2 replicates) 2) 480 million 86-nucleotide reads were generated from four lanes of Illuminia HiSeqII 3) 74% of the reads could be mapped to the sorghum transcriptome, indicative of good sequence quality 4) Of the 216 genes within the QTL, 17 genes were differentially expressed among plants with and without the Rio QTL. None of these 17 genes had obvious roles in sucrose metabolism 5) Clustering algorithms identified a group of 721 co-expressed genes. One of these genes is a sucrose synthase gene. This cluster also contains 10 genes from the QTL. 6) Among these co-expressed genes are regulatory genes for which knock-out lines in Arabidopsis have been obtained. Analysis of

  15. Solid-phase fermentation and juice expression systems for sweet sorghum

    Energy Technology Data Exchange (ETDEWEB)

    Bryan, W.L.; Monroe, G.E.; Caussariel, P.M.

    1985-01-01

    Two systems to recover fermented juice from variety M 81E sweet sorghum stalks that contained about 11% fermentable sugar were compared. (a) Stalks with leaves and tops removed were chopped and inoculated with 0.2% yeast in a forage harvester, stored under anaerobic conditions for 75 hours in insulated fermentors and pressed in a screw press to recover fermented juice (5-6% ethanol). (b) Mechanically harvested sweet sorghum billets (30 cm length) without leaves or seed heads were shredded and milled in a 3-roll mill; and bagasse was inoculated with 0.2% yeast, fermented for 100 h and pressed to recover fermented juice (4 to 5% ethanol). Potential ethanol yields were 75% of theoretical for the forage harvest system and 78% for the shredder mill system, based on 95% of theoretical ethanol yield from juice expressed during milling and no loss of ethanol during fermentation, handling and pressing in the screw press. 20 references.

  16. Cloning, recombinant expression and characterization of a new ...

    African Journals Online (AJOL)

    A new amylase gene APGA1 was cloned from Aureobasidium pullulans NRRL 12974 and expressed in Pichia pastoris. This is the first report on cloning and expression of amylolytic gene from the industrially important microorganism A. pullulans. The purified recombinant protein with MW of 66 kDa and specific activity of ...

  17. Molecular and phenotypic characterization of transgenic wheat and sorghum events expressing the barley alanine aminotransferase.

    Science.gov (United States)

    Peña, Pamela A; Quach, Truyen; Sato, Shirley; Ge, Zhengxiang; Nersesian, Natalya; Dweikat, Ismail M; Soundararajan, Madhavan; Clemente, Tom

    2017-12-01

    The expression of a barley alanine aminotransferase gene impacts agronomic outcomes in a C3 crop, wheat. The use of nitrogen-based fertilizers has become one of the major agronomic inputs in crop production systems. Strategies to enhance nitrogen assimilation and flux in planta are being pursued through the introduction of novel genetic alleles. Here an Agrobacterium-mediated approach was employed to introduce the alanine aminotransferase from barley (Hordeum vulgare), HvAlaAT, into wheat (Triticum aestivum) and sorghum (Sorghum bicolor), regulated by either constitutive or root preferred promoter elements. Plants harboring the transgenic HvAlaAT alleles displayed increased alanine aminotransferase (alt) activity. The enhanced alt activity impacted height, tillering and significantly boosted vegetative biomass relative to controls in wheat evaluated under hydroponic conditions, where the phenotypic outcome across these parameters varied relative to time of year study was conducted. Constitutive expression of HvAlaAT translated to elevation in wheat grain yield under field conditions. In sorghum, expression of HvAlaAT enhanced enzymatic activity, but no changes in phenotypic outcomes were observed. Taken together these results suggest that positive agronomic outcomes can be achieved through enhanced alt activity in a C3 crop, wheat. However, the variability observed across experiments under greenhouse conditions implies the phenotypic outcomes imparted by the HvAlaAT allele in wheat may be impacted by environment.

  18. [TSA improve transgenic porcine cloned embryo development and transgene expression].

    Science.gov (United States)

    Kong, Qing-Ran; Zhu, Jiang; Huang, Bo; Huan, Yan-Jun; Wang, Feng; Shi, Yong-Qian; Liu, Zhong-Feng; Wu, Mei-Ling; Liu, Zhong-Hua

    2011-07-01

    Uncompleted epigenetic reprogramming is attributed to the low efficiency of producing transgenic cloned animals. Histone modification associated with epigenetics can directly influence the embryo development and transgene expression. Trichostatin A (TSA), as an inhibitor of histone deacetylase, can change the status of histone acetylation, improve somatic cell reprogramming, and enhance cloning efficiency. TSA prevents the chromatin structure from being condensed, so that transcription factor could binds to DNA sequence easily and enhance transgene expression. Our study established the optimal TSA treatment on porcine donor cells and cloned embryos, 250 nmol/L, 24 h and 40 nmol/L, 24 h, respectively. Furthermore, we found that both the cloned embryo and the donor cell treated by TSA resulted in the highest development efficiency. Meanwhile, TSA can improve transgene expression in donor cell and cloned embryo. In summary, TSA can significantly improve porcine reconstructed embryo development and transgene expression.

  19. Achievements and problems in the weed control in grain sorghum (Sorghum Bicolor Moench.

    Directory of Open Access Journals (Sweden)

    Gr. Delchev

    2017-09-01

    Full Text Available Abstract. Chemical control has emerged as the most efficient method of weed control. Herbicides combinations and tank mixtures of herbicides with adjuvants, fertilizers, growth regulators, fungicides, insecticides are more effective than when applied alone on sorghum crops. Their combined use often leads to high synergistic effect on yield. The use of herbicide antidotes for the treatment of seeds in sorghum is a safe way to overcome its high sensitivity to many herbicides. Data regarding herbicide for chemical control of annual graminaceous weeds in sorghum crops are quite scarce even worldwide. Problem is the persistence of some herbicides used in the predecessors on succeeding crops, which is directly related to the weather conditions during their degradation. Most of the information on sorghum relates to the conventional technology for weed control. There is no information about the new Concep technology in grain sorghum. A serious problem is also the volunteers of the Clearfield and Express sun sunflower. They have resistance to herbicides different from that of conventional sunflower hybrids. There is no information yet in scientific literature on control of these volunteers.

  20. Differential expression of two flavonoid 3'-hydroxylase cDNAs involved in biosynthesis of anthocyanin pigments and 3-deoxyanthocyanidin phytoalexins in sorghum.

    Science.gov (United States)

    Shih, Chun-Hat; Chu, Ivan K; Yip, Wing Kin; Lo, Clive

    2006-10-01

    Three unique sorghum flavonoid 3'-hydroxylase (F3'H) cDNAs (SbF3'H1, SbF3'H2 and SbF3'H3) were discovered through bioinformatics analysis. Their encoded proteins showed >60% identity to the Arabidopsis TT7 (F3'H) protein. Overexpression of SbF3'H1 or SbF3'H2 restored the ability of tt7 mutants to produce 3'-hydroxylated flavonoids, establishing their roles as functional F3'H enzymes. In sorghum mesocotyls, SbF3'H1 expression was involved in light-specific anthocyanin accumulation while SbF3'H2 expression was involved in pathogen-specific 3-deoxyanthocyanidin synthesis. No SbF3'H3 expression was detected in all tissues examined. The sorghum mesocotyls represent a good system for investigation of differential regulation of F3'H genes/alleles responding to different external stimuli.

  1. TA-GC cloning: A new simple and versatile technique for the directional cloning of PCR products for recombinant protein expression.

    Science.gov (United States)

    Niarchos, Athanasios; Siora, Anastasia; Konstantinou, Evangelia; Kalampoki, Vasiliki; Lagoumintzis, George; Poulas, Konstantinos

    2017-01-01

    During the last few decades, the recombinant protein expression finds more and more applications. The cloning of protein-coding genes into expression vectors is required to be directional for proper expression, and versatile in order to facilitate gene insertion in multiple different vectors for expression tests. In this study, the TA-GC cloning method is proposed, as a new, simple and efficient method for the directional cloning of protein-coding genes in expression vectors. The presented method features several advantages over existing methods, which tend to be relatively more labour intensive, inflexible or expensive. The proposed method relies on the complementarity between single A- and G-overhangs of the protein-coding gene, obtained after a short incubation with T4 DNA polymerase, and T and C overhangs of the novel vector pET-BccI, created after digestion with the restriction endonuclease BccI. The novel protein-expression vector pET-BccI also facilitates the screening of transformed colonies for recombinant transformants. Evaluation experiments of the proposed TA-GC cloning method showed that 81% of the transformed colonies contained recombinant pET-BccI plasmids, and 98% of the recombinant colonies expressed the desired protein. This demonstrates that TA-GC cloning could be a valuable method for cloning protein-coding genes in expression vectors.

  2. Sorghum allelopathy--from ecosystem to molecule.

    Science.gov (United States)

    Weston, Leslie A; Alsaadawi, Ibrahim S; Baerson, Scott R

    2013-02-01

    Sorghum allelopathy has been reported in a series of field experiments following sorghum establishment. In recent years, sorghum phytotoxicity and allelopathic interference also have been well-described in greenhouse and laboratory settings. Observations of allelopathy have occurred in diverse locations and with various sorghum plant parts. Phytotoxicity has been reported when sorghum was incorporated into the soil as a green manure, when residues remained on the soil surface in reduced tillage settings, or when sorghum was cultivated as a crop in managed fields. Allelochemicals present in sorghum tissues have varied with plant part, age, and cultivar evaluated. A diverse group of sorghum allelochemicals, including numerous phenolics, a cyanogenic glycoside (dhurrin), and a hydrophobic p-benzoquinone (sorgoleone) have been isolated and identified in recent years from sorghum shoots, roots, and root exudates, as our capacity to analyze and identify complex secondary products in trace quantities in the plant and in the soil rhizosphere has improved. These allelochemicals, particularly sorgoleone, have been widely investigated in terms of their mode(s) of action, specific activity and selectivity, release into the rhizosphere, and uptake and translocation into sensitive indicator species. Both genetics and environment have been shown to influence sorgoleone production and expression of genes involved in sorgoleone biosynthesis. In the soil rhizosphere, sorgoleone is released continuously by living root hairs where it accumulates in significant concentrations around its roots. Further experimentation designed to study the regulation of sorgoleone production by living sorghum root hairs may result in increased capacity to utilize sorghum cover crops more effectively for suppression of germinating weed seedlings, in a manner similar to that of soil-applied preemergent herbicides like trifluralin.

  3. TA-GC cloning: A new simple and versatile technique for the directional cloning of PCR products for recombinant protein expression.

    Directory of Open Access Journals (Sweden)

    Athanasios Niarchos

    Full Text Available During the last few decades, the recombinant protein expression finds more and more applications. The cloning of protein-coding genes into expression vectors is required to be directional for proper expression, and versatile in order to facilitate gene insertion in multiple different vectors for expression tests. In this study, the TA-GC cloning method is proposed, as a new, simple and efficient method for the directional cloning of protein-coding genes in expression vectors. The presented method features several advantages over existing methods, which tend to be relatively more labour intensive, inflexible or expensive. The proposed method relies on the complementarity between single A- and G-overhangs of the protein-coding gene, obtained after a short incubation with T4 DNA polymerase, and T and C overhangs of the novel vector pET-BccI, created after digestion with the restriction endonuclease BccI. The novel protein-expression vector pET-BccI also facilitates the screening of transformed colonies for recombinant transformants. Evaluation experiments of the proposed TA-GC cloning method showed that 81% of the transformed colonies contained recombinant pET-BccI plasmids, and 98% of the recombinant colonies expressed the desired protein. This demonstrates that TA-GC cloning could be a valuable method for cloning protein-coding genes in expression vectors.

  4. Insights into the Bamboo Genome: Syntenic Relationships to Rice and Sorghum

    Institute of Scientific and Technical Information of China (English)

    Yi-Jie Gui; Nai-Xun Ma; Tian-Zhen Zhang; Long-Jiang Fan; Yan Zhou; Yu Wang; Sheng Wang; Sheng-Yue Wang; Yan Hu; Shi-Ping Bo; Huan Chen; Chang-Ping Zhou

    2010-01-01

    Bamboo occupies an important phylogenetic node in the grass family and plays a significant role in the forest industry.We produced 1.2 Mb of tetraploid moso bamboo(Phyllostachys pubescens E.Mazel ex H.de Leh.)sequences from 13 bacterial artificial chromosome(BAC)clones,and these are the largest genomic sequences available so far from the subfamily Bambusoideae.The content of repetitive elements(36.2%)in bamboo is similar to that in rice.Both rice and sorghum exhibit high genomic synteny with bamboo,which suggests that rice and sorghum may be useful as models for decoding Bambusoideae genomes.

  5. Aberrant epigenetic changes and gene expression in cloned cattle dying around birth

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    Zhao Dingsheng

    2008-02-01

    Full Text Available Abstract Background Aberrant reprogramming of donor somatic cell nuclei may result in many severe problems in animal cloning. To assess the extent of abnormal epigenetic modifications and gene expression in clones, we simultaneously examined DNA methylation, histone H4 acetylation and expression of six genes (β-actin, VEGF, oct4, TERT, H19 and Igf2 and a repetitive sequence (art2 in five organs (heart, liver, spleen, lung and kidney from two cloned cattle groups that had died at different stages. In the ED group (early death, n = 3, the cloned cattle died in the perinatal period. The cattle in the LD group (late death, n = 3 died after the perinatal period. Normally reproduced cattle served as a control group (n = 3. Results Aberrant DNA methylation, histone H4 acetylation and gene expression were observed in both cloned groups. The ED group showed relatively fewer severe DNA methylation abnormalities (p Conclusion Deaths of clones may be ascribed to abnormal expression of a very limited number of genes.

  6. Frame-Insensitive Expression Cloning of Fluorescent Protein from Scolionema suvaense

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    Yuki Horiuchi

    2018-01-01

    Full Text Available Expression cloning from cDNA is an important technique for acquiring genes encoding novel fluorescent proteins. However, the probability of in-frame cDNA insertion following the first start codon of the vector is normally only 1/3, which is a cause of low cloning efficiency. To overcome this issue, we developed a new expression plasmid vector, pRSET-TriEX, in which transcriptional slippage was induced by introducing a DNA sequence of (dT14 next to the first start codon of pRSET. The effectiveness of frame-insensitive cloning was validated by inserting the gene encoding eGFP with all three possible frames to the vector. After transformation with one of these plasmids, E. coli cells expressed eGFP with no significant difference in the expression level. The pRSET-TriEX vector was then used for expression cloning of a novel fluorescent protein from Scolionema suvaense. We screened 3658 E. coli colonies transformed with pRSET-TriEX containing Scolionema suvaense cDNA, and found one colony expressing a novel green fluorescent protein, ScSuFP. The highest score in protein sequence similarity was 42% with the chain c of multi-domain green fluorescent protein like protein “ember” from Anthoathecata sp. Variations in the N- and/or C-terminal sequence of ScSuFP compared to other fluorescent proteins indicate that the expression cloning, rather than the sequence similarity-based methods, was crucial for acquiring the gene encoding ScSuFP. The absorption maximum was at 498 nm, with an extinction efficiency of 1.17 × 105 M−1·cm−1. The emission maximum was at 511 nm and the fluorescence quantum yield was determined to be 0.6. Pseudo-native gel electrophoresis showed that the protein forms obligatory homodimers.

  7. Genome analysis methods: Sorghum bicolor [PGDBj Registered plant list, Marker list, QTL list, Plant DB link and Genome analysis methods[Archive

    Lifescience Database Archive (English)

    Full Text Available Sorghum bicolor Finished 2n=20 760 Mb 2009 Sanger (Clone-based) 10,717,203 reads 7...30 Mb 8.5x Arachne2 v.20060705 3,304 12,873 BLAST, GenomeScan 34,496 (Sbi1.4) JGI; http://www.phytozome.net/sorghum Sbi1 Sbi1.4 10.1038/nature07723 19189423 ...

  8. Combining ability and mode of inheritance of stem thickness in forage sorghum (Sorghum bicolor L. Moench F1 hybrids

    Directory of Open Access Journals (Sweden)

    Pataki Imre

    2011-01-01

    Full Text Available Aim of this research was determination of mode of inheritance, gene effects components of genetic variance, combining abilities, average contribution of lines and testers and their interactions in expression of stem thickness in forage sorghum F1 generation. Method line x tester was applied. Material comprised of eight genetically divergent A-inbred lines of grain sorghum three R lines-testers of Sudan grass and twenty-four F1 hybrids obtained by crossing lines with testers. Among tested genotypes there were significant differences in mean values of stem thickness. Analysis of variance of combining abilities showed that there were highly significant differences for general combining abilities (GCA and specific combining abilities (SCA non-additive component of genetic variance (dominance and epistasis had greater portion in total genetic variance for stem thickness. During the first research year, interaction between inbred maternal line with testers had the largest contribution in expression of stem thickness of F1 hybrid at both locations, while in the second year at location Rimski Šančevi the largest contribution belongs to lines and at location Mačvanski Prnjavor the largest contribution belongs to testers. Assessment of combining abilities showed that these inbred lines of grain sorghum can be used as mothers: SS-1 646, SS-1 688 and S-8 682 in breeding forage sorghum for thicker stem. According to SCA, promising forage sorghum hybrids are S-8 682 x ST-R lin H and P-21 656 x C-198. This research can be of importance for developing new high-yielding forage sorghum hybrids.

  9. Transgenic sorghum ( Sorghum bicolor L. Moench) developed by ...

    African Journals Online (AJOL)

    Sorghum (Sorghum bicolor (L.) Moench) is an important food and fodder crop. Fungal diseases such as anthracnose caused by Colletotrichum sublineolum reduce sorghum yields. Genetic transformation can be used to confer tolerance to plant diseases such as anthracnose. The tolerance can be developed by introducing ...

  10. Cloning, expression, and chromosome mapping of human galectin-7

    DEFF Research Database (Denmark)

    Madsen, Peder; Rasmussen, H H; Flint, T

    1995-01-01

    The galectins are a family of beta-galactoside-binding proteins implicated in modulating cell-cell and cell-matrix interactions. Here we report the cloning and expression of a novel member of this family (galectin-7) that correspond to IEF (isoelectric focusing) 17 (12,700 Da; pI, 7.6) in the human...... keratinocyte protein data base, and that is strikingly down-regulated in SV40 transformed keratinocytes (K14). The cDNA was cloned from a lambda gt11 cDNA expression library using degenerated oligodeoxyribonucleotides back-translated from an IEF 17 peptide sequence. The protein encoded by the galectin-7 clone......14 keratinocytes imply a role in cell-cell and/or cell-matrix interactions necessary for normal growth control. The galectin-7 gene was mapped to chromosome 19. Udgivelsesdato: 1995-Mar-17...

  11. Cloning, sequencing, and expression of cDNA for human β-glucuronidase

    International Nuclear Information System (INIS)

    Oshima, A.; Kyle, J.W.; Miller, R.D.

    1987-01-01

    The authors report here the cDNA sequence for human placental β-glucuronidase (β-D-glucuronoside glucuronosohydrolase, EC 3.2.1.31) and demonstrate expression of the human enzyme in transfected COS cells. They also sequenced a partial cDNA clone from human fibroblasts that contained a 153-base-pair deletion within the coding sequence and found a second type of cDNA clone from placenta that contained the same deletion. Nuclease S1 mapping studies demonstrated two types of mRNAs in human placenta that corresponded to the two types of cDNA clones isolated. The NH 2 -terminal amino acid sequence determined for human spleen β-glucuronidase agreed with that inferred from the DNA sequence of the two placental clones, beginning at amino acid 23, suggesting a cleaved signal sequence of 22 amino acids. When transfected into COS cells, plasmids containing either placental clone expressed an immunoprecipitable protein that contained N-linked oligosaccharides as evidenced by sensitivity to endoglycosidase F. However, only transfection with the clone containing the 153-base-pair segment led to expression of human β-glucuronidase activity. These studies provide the sequence for the full-length cDNA for human β-glucuronidase, demonstrate the existence of two populations of mRNA for β-glucuronidase in human placenta, only one of which specifies a catalytically active enzyme, and illustrate the importance of expression studies in verifying that a cDNA is functionally full-length

  12. Molecular Cloning Expression And Purification Studies With An ORF Of Mycobacterium Tuberculosis

    Directory of Open Access Journals (Sweden)

    Chiranjibi Chaudhary

    2017-08-01

    Full Text Available The study was initiated to develop a recombinant strain for expression and production of large scale protein and to develop its purification protocol. The MRAORF-X was amplified from the genomic DNA of M. tuberculosis H37Ra. The amplicon was successfully cloned in a cloning vector pGEM-T Easy and transformed in cloning host DH5amp945. Recombinant clones were identified by blue-white screening and insert presence was confirmed by restriction digestion of plasmid isolated from white colonies. Expression vector pET32a was used for protein expression. The recombinant plasmid was transformed into expression host BL21 and protein expression was checked by SDS-PAGE. The desired protein was approximately 60 kDa in size including tags. The purification protocol was established for purification from inclusion bodies. The purity of purified protein was assessed by SDS-PAGE gel run and presence of a single band at 60 kDa suggested that the inclusion bodies were a good source of purified protein.

  13. Production of cloned pigs with targeted attenuation of gene expression.

    Directory of Open Access Journals (Sweden)

    Vilceu Bordignon

    Full Text Available The objective of this study was to demonstrate that RNA interference (RNAi and somatic cell nuclear transfer (SCNT technologies can be used to attenuate the expression of specific genes in tissues of swine, a large animal species. Apolipoprotein E (apoE, a secreted glycoprotein known for its major role in lipid and lipoprotein metabolism and transport, was selected as the target gene for this study. Three synthetic small interfering RNAs (siRNA targeting the porcine apoE mRNA were tested in porcine granulosa cells in primary culture and reduced apoE mRNA abundance ranging from 45-82% compared to control cells. The most effective sequence was selected for cloning into a short hairpin RNA (shRNA expression vector under the control of RNA polymerase III (U6 promoter. Stably transfected fetal porcine fibroblast cells were generated and used to produce embryos with in vitro matured porcine oocytes, which were then transferred into the uterus of surrogate gilts. Seven live and one stillborn piglet were born from three gilts that became pregnant. Integration of the shRNA expression vector into the genome of clone piglets was confirmed by PCR and expression of the GFP transgene linked to the expression vector. Analysis showed that apoE protein levels in the liver and plasma of the clone pigs bearing the shRNA expression vector targeting the apoE mRNA was significantly reduced compared to control pigs cloned from non-transfected fibroblasts of the same cell line. These results demonstrate the feasibility of applying RNAi and SCNT technologies for introducing stable genetic modifications in somatic cells for eventual attenuation of gene expression in vivo in large animal species.

  14. Three sorghum serpin recombinant proteins inhibit midgut trypsin activity and growth of corn earworm

    Science.gov (United States)

    The sorghum (Sorghum bicolor) genome contains at least 17 putative serpin (serine protease inhibitor) open reading frames, some of which are induced by pathogens. Recent transcriptome studies found that most of the putative serpins are expressed but their roles are unknown. Four sorghum serpins were...

  15. The wheat Lr34 multipathogen resistance gene confers resistance to anthracnose and rust in sorghum.

    Science.gov (United States)

    Schnippenkoetter, Wendelin; Lo, Clive; Liu, Guoquan; Dibley, Katherine; Chan, Wai Lung; White, Jodie; Milne, Ricky; Zwart, Alexander; Kwong, Eunjung; Keller, Beat; Godwin, Ian; Krattinger, Simon G; Lagudah, Evans

    2017-11-01

    The ability of the wheat Lr34 multipathogen resistance gene (Lr34res) to function across a wide taxonomic boundary was investigated in transgenic Sorghum bicolor. Increased resistance to sorghum rust and anthracnose disease symptoms following infection with the biotrophic pathogen Puccinia purpurea and the hemibiotroph Colletotrichum sublineolum, respectively, occurred in transgenic plants expressing the Lr34res ABC transporter. Transgenic sorghum lines that highly expressed the wheat Lr34res gene exhibited immunity to sorghum rust compared to the low-expressing single copy Lr34res genotype that conferred partial resistance. Pathogen-induced pigmentation mediated by flavonoid phytoalexins was evident on transgenic sorghum leaves following P. purpurea infection within 24-72 h, which paralleled Lr34res gene expression. Elevated expression of flavone synthase II, flavanone 4-reductase and dihydroflavonol reductase genes which control the biosynthesis of flavonoid phytoalexins characterized the highly expressing Lr34res transgenic lines 24-h post-inoculation with P. purpurea. Metabolite analysis of mesocotyls infected with C. sublineolum showed increased levels of 3-deoxyanthocyanidin metabolites were associated with Lr34res expression, concomitant with reduced symptoms of anthracnose. © 2017 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

  16. The application of secondary metabolites in the study of sorghum insect resistance

    Science.gov (United States)

    Chunming, Bai; Yifei, Liu; Xiaochun, Lu

    2018-03-01

    Insect attack is one of the main factors for limiting the production of rice and sorghum. To improve resistance to pests of rice and sorghum will be of great significance for meliorating their production and quality. However, the source and material of anti-pest was scarce. In this study, we will study on the expression patterns of hydrocyanic acid biosynthesis relative genes in sorghum firstly. And we will also genetically transform them into rice and sorghum by specific and constitutive promoters and verify their pest-resistant ability. Finally, high pest-resistant genetically modified new sorghum cultivars will be bred with favorable comprehensive agronomic traits.

  17. Introduction of sorghum (Sorghum bicolor (L.) Moench) into China ...

    African Journals Online (AJOL)

    The sorghum is a plant, which has been intentionally introduced in China for foods needs. It is a plant of African origin, which is much cultivated in the northern hemisphere. For millions of people in the semiarid tropic temperature of Asia and Africa, sorghum is the most important staple food. Sorghum is becoming one of the ...

  18. Identification, Characterization and Expression Analysis of Cell Wall Related Genes in Sorghum bicolor (L. Moench, a Food, Fodder and Biofuel Crop

    Directory of Open Access Journals (Sweden)

    KRISHAN MOHAN RAI

    2016-08-01

    Full Text Available Biomass based alternative fuels offer a solution to the world’s ever-increasing energy demand. With the ability to produce high biomass in marginal lands with low inputs, sorghum has a great potential to meet second-generation biofuel needs. Despite the sorghum crop importance in biofuel and fodder industry, there is no comprehensive information available on the cell wall related genes and gene families (biosynthetic and modification. It is important to identify the cell wall related genes to understand the cell wall biosynthetic process as well as to facilitate biomass manipulation. Genome-wide analysis using gene family specific Hidden Markov Model of conserved domains identified 520 genes distributed among 20 gene families related to biosynthesis/modification of various cell wall polymers such as cellulose, hemicellulose, pectin and lignin. Chromosomal localization analysis of these genes revealed that about 65% of cell wall related genes were confined to four chromosomes (Chr. 1-4. Further, 53 tandem duplication events involving 146 genes were identified in these gene families which could be associated with expansion of genes within families in sorghum. Additionally, we also identified 137 Simple Sequence Repeats related to 112 genes and target sites for 10 miRNAs in some important families such as cellulose synthase, cellulose synthase-like and laccases, etc. To gain further insight into potential functional roles, expression analysis of these gene families was performed using publicly available data sets in various tissues and under abiotic stress conditions. Expression analysis showed tissue specificity as well as differential expression under abiotic stress conditions. Overall, our study provides a comprehensive information on cell wall related genes families in sorghum which offers a valuable resource to develop strategies for altering biomass composition by plant breeding and genetic engineering approaches.

  19. Cloning an expressed gene shared by the human sex chromosomes

    International Nuclear Information System (INIS)

    Darling, S.M.; Banting, G.S.; Pym, B.; Wolfe, J.; Goodfellow, P.N.

    1986-01-01

    The existence of genes shared by mammalian sex chromosomes has been predicted on both evolutionary and functional grounds. However, the only experimental evidence for such genes in humans is the cell-surface antigen encoded by loci on the X and Y chromosomes (MIC2X and MIC2Y, respectively), which is recognized by the monoclonal antibody 12E7. Using the bacteriophage λgt11 expression system in Escherichia coli and immunoscreening techniques, the authors have isolated a cDNA clone whose primary product is recognized by 12E7. Southern blot analysis using somatic cell hybrids containing only the human X or Y chromosomes shows that the sequences reacting with the cDNA clone are localized to the sex chromosomes. In addition, the clone hybridizes to DNAs isolated from mouse cells that have been transfected with human DNA and selected for 12E7 expression on the fluorescence-activated cell sorter. The authors conclude that the cDNA clone encodes the 12E7 antigen, which is the primary product of the MIC2 loci. The clone was used to explore sequence homology between MIC2X and MIC2Y; these loci are closely related, if not identical

  20. Recombinational Cloning Using Gateway and In-Fusion Cloning Schemes

    Science.gov (United States)

    Throop, Andrea L.; LaBaer, Joshua

    2015-01-01

    The comprehensive study of protein structure and function, or proteomics, depends on the obtainability of full-length cDNAs in species-specific expression vectors and subsequent functional analysis of the expressed protein. Recombinational cloning is a universal cloning technique based on site-specific recombination that is independent of the insert DNA sequence of interest, which differentiates this method from the classical restriction enzyme-based cloning methods. Recombinational cloning enables rapid and efficient parallel transfer of DNA inserts into multiple expression systems. This unit summarizes strategies for generating expression-ready clones using the most popular recombinational cloning technologies, including the commercially available Gateway® (Life Technologies) and In-Fusion® (Clontech) cloning technologies. PMID:25827088

  1. High-Throughput Cloning and Expression Library Creation for Functional Proteomics

    Science.gov (United States)

    Festa, Fernanda; Steel, Jason; Bian, Xiaofang; Labaer, Joshua

    2013-01-01

    The study of protein function usually requires the use of a cloned version of the gene for protein expression and functional assays. This strategy is particular important when the information available regarding function is limited. The functional characterization of the thousands of newly identified proteins revealed by genomics requires faster methods than traditional single gene experiments, creating the need for fast, flexible and reliable cloning systems. These collections of open reading frame (ORF) clones can be coupled with high-throughput proteomics platforms, such as protein microarrays and cell-based assays, to answer biological questions. In this tutorial we provide the background for DNA cloning, discuss the major high-throughput cloning systems (Gateway® Technology, Flexi® Vector Systems, and Creator™ DNA Cloning System) and compare them side-by-side. We also report an example of high-throughput cloning study and its application in functional proteomics. This Tutorial is part of the International Proteomics Tutorial Programme (IPTP12). Details can be found at http://www.proteomicstutorials.org. PMID:23457047

  2. High-throughput Cloning and Expression of Integral Membrane Proteins in Escherichia coli

    Science.gov (United States)

    Bruni, Renato

    2014-01-01

    Recently, several structural genomics centers have been established and a remarkable number of three-dimensional structures of soluble proteins have been solved. For membrane proteins, the number of structures solved has been significantly trailing those for their soluble counterparts, not least because over-expression and purification of membrane proteins is a much more arduous process. By using high throughput technologies, a large number of membrane protein targets can be screened simultaneously and a greater number of expression and purification conditions can be employed, leading to a higher probability of successfully determining the structure of membrane proteins. This unit describes the cloning, expression and screening of membrane proteins using high throughput methodologies developed in our laboratory. Basic Protocol 1 deals with the cloning of inserts into expression vectors by ligation-independent cloning. Basic Protocol 2 describes the expression and purification of the target proteins on a miniscale. Lastly, for the targets that express at the miniscale, basic protocols 3 and 4 outline the methods employed for the expression and purification of targets at the midi-scale, as well as a procedure for detergent screening and identification of detergent(s) in which the target protein is stable. PMID:24510647

  3. Cloning and Expression Vector Construction of Glutamate Decarboxylase Gene from Lactobacillus Plantarum

    Directory of Open Access Journals (Sweden)

    B Arabpour

    2016-06-01

    Full Text Available BACKGROUND AND OBJECTIVE: Gamma-aminobutyric acid (GABA is a four-carbon non-protein amino acid used in the treatment of hypertension, diabetes, inflammation, and depression. GABA is synthesized by glutamic acid decarboxylase (GAD enzyme in many organisms, including bacteria. Therefore, cloning of this enzyme is essential to the optimization of GABA production. This study aimed to clone and construct the expression vector of GAD gene from Lactobacillus plantarum PTCC 1058 bacterium. METHODS: In this experimental study, we investigated the morphological, biochemical, genetic and 16s rDNA sequencing of L. plantarum PTCC 1058 strain. Genomic DNA of the bacterium was isolated and amplified using the GAD gene via polymerase chain reaction (PCR. Afterwards, the gene was inserted into the pJET1.2/blunt cloning vector and subcloned in vector pET32a. Plasmid pET32a-gad expression vector was transformed in Escherichia coli BL21 strain, and protein expression was assessed using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE. FINDINGS: Morphological, biochemical and genetic analyses of 16s rDNA sequencing indicated that the studied substrain was of the L. plantarum strain. In addition, results of nucleotide sequencing of the fragmented segment via PCR showed the presence of GAD gene. Results of colony PCR and SDS-PAGE analysis confirmed the accuracy of the cloning and gene expression of the recombinant Escherichia coli BL21 strain. CONCLUSION: According to the results of this study, cloning of GAD gene from L. plantarum PTCC 1058 was successful. These cloned genes could grow rapidly in prokaryotic and eukaryotic systems and be used in cost-effective culture media and even non-recyclable waste.

  4. Characterization of Nitrogen use efficiency in sweet sorghum

    Energy Technology Data Exchange (ETDEWEB)

    Dweikat, Ismail [University of Nebraska; Clemente, Thomas [University of Nebrask

    2014-09-09

    Sweet sorghum (Sorghum bicolor L. Moench) has the potential to augment the increasing demand for alternative fuels and for the production of input efficient, environmentally friendly bioenergy crops. Nitrogen (N) and water availability are considered two of the major limiting factors in crop growth. Nitrogen fertilization accounts for about 40% of the total production cost in sorghum. In cereals, including sorghum, the nitrogen use efficiency (NUE) from fertilizer is approximately 33% of the amount applied. There is therefore extensive concern in relation to the N that is not used by the plant, which is lost by leaching of nitrate, denitrification from the soil, and loss of ammonia to the atmosphere, all of which can have deleterious environmental effects. To improve the potential of sweet sorghum as a leading and cost effective bioenergy crop, the enhancement of NUE must be addressed. To this end, we have identified a sorghum line (SanChi San) that displays about 25% increase in NUE over other sorghum lines. As such, the overarching goal of this project is to employ three complementary strategies to enhance the ability of sweet sorghum to become an efficient nitrogen user. To achieve the project goal, we will pursue the following specific objectives: Objective 1: Phenotypic characterization of SanChi San/Ck60 RILs under low and moderate N-availability including biochemical profiles, vegetative growth and seed yield Objective 2: Conduct quantitative trait loci (QTL) analysis and marker identification for nitrogen use efficiency (NUE) in a grain sorghum RIL population. Objective 3: Identify novel candidate genes for NUE using proteomic and gene expression profiling comparisons of high- and low-NUE RILs. Candidate genes will be brought into the pipeline for transgenic manipulation of NUE This project will apply the latest genomics resources to discover genes controlling NUE, one of the most complex and economically important traits in cereal crops. As a result of the

  5. Variation in gene expression within clones of the earthworm Dendrobaena octaedra.

    Directory of Open Access Journals (Sweden)

    Marina Mustonen

    Full Text Available Gene expression is highly plastic, which can help organisms to both acclimate and adapt to changing environments. Possible variation in gene expression among individuals with the same genotype (among clones is not widely considered, even though it could impact the results of studies that focus on gene expression phenotypes, for example studies using clonal lines. We examined the extent of within and between clone variation in gene expression in the earthworm Dendrobaena octaedra, which reproduces through apomictic parthenogenesis. Five microsatellite markers were developed and used to confirm that offspring are genetic clones of their parent. After that, expression of 12 genes was measured from five individuals each from six clonal lines after exposure to copper contaminated soil. Variation in gene expression was higher over all genotypes than within genotypes, as initially assumed. A subset of the genes was also examined in the offspring of exposed individuals in two of the clonal lines. In this case, variation in gene expression within genotypes was as high as that observed over all genotypes. One gene in particular (chymotrypsin inhibitor also showed significant differences in the expression levels among genetically identical individuals. Gene expression can vary considerably, and the extent of variation may depend on the genotypes and genes studied. Ensuring a large sample, with many different genotypes, is critical in studies comparing gene expression phenotypes. Researchers should be especially cautious inferring gene expression phenotypes when using only a single clonal or inbred line, since the results might be specific to only certain genotypes.

  6. Enzyme free cloning for high throughput gene cloning and expression

    NARCIS (Netherlands)

    de Jong, R.N.; Daniëls, M.; Kaptein, R.; Folkers, G.E.

    2006-01-01

    Structural and functional genomics initiatives significantly improved cloning methods over the past few years. Although recombinational cloning is highly efficient, its costs urged us to search for an alternative high throughput (HTP) cloning method. We implemented a modified Enzyme Free Cloning

  7. [Cloning of human CD45 gene and its expression in Hela cells].

    Science.gov (United States)

    Li, Jie; Xu, Tianyu; Wu, Lulin; Zhang, Liyun; Lu, Xiao; Zuo, Daming; Chen, Zhengliang

    2015-11-01

    To clone human CD45 gene PTPRC and establish Hela cells overexpressing recombinant human CD45 protein. The intact cDNA encoding human CD45 amplified using RT-PCR from the total RNA extracted from peripheral blood mononuclear cells (PBMCs) of a healthy donor was cloned into pMD-18T vector. The CD45 cDNA fragment amplified from the pMD-18T-CD45 by PCR was inserted to the coding region of the PcDNA3.1-3xflag vector, and the resultant recombinant expression vector PcDNA3.1-3xflag-CD45 was transfected into Hela cells. The expression of CD45 in Hela cells was detected by flow cytometry and Western blotting, and the phosphastase activity of CD45 was quantified using an alkaline phosphatase assay kit. The cDNA fragment of about 3 900 bp was amplified from human PBMCs and cloned into pMD-18T vector. The recombinant expression vector PcDNA3.1-3xflag-CD45 was constructed, whose restriction maps and sequence were consistent with those expected. The expression of CD45 in transfected Hela cells was detected by flow cytometry and Western blotting, and the expressed recombinant CD45 protein in Hela cells showed a phosphastase activity. The cDNA of human CD45 was successfully cloned and effectively expressed in Hela cells, which provides a basis for further exploration of the functions of CD45.

  8. Cloning and semi-quantitative expression of endochitinase ( ech42 ...

    African Journals Online (AJOL)

    Cloning and semi-quantitative expression of endochitinase (ech42) gene from Trichoderma spp. Pratibha Sharma, K Saravanan, R Ramesh, P Vignesh Kumar, Dinesh Singh, Manika Sharma, Monica S. Henry, Swati Deep ...

  9. Taxonomy Icon Data: sorghum [Taxonomy Icon

    Lifescience Database Archive (English)

    Full Text Available sorghum Sorghum bicolor Sorghum_bicolor_L.png Sorghum_bicolor_NL.png Sorghum_bicolor_S.png Sorg...hum_bicolor_NS.png http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Sorghum+bicolor&t=L http://b...iosciencedbc.jp/taxonomy_icon/icon.cgi?i=Sorghum+bicolor&t=NL http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Sorg...hum+bicolor&t=S http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Sorghum+bicolor&t=NS ...

  10. Advances in sorghum genetic mapping with implications for sorghum improvement

    International Nuclear Information System (INIS)

    Lee, M.

    1998-01-01

    Despite the importance of the sorghum crop, comprehensive genetic characterization has been limited. Therefore, the primary goal of this research program was to develop basic genetic tools to facilitate research in the genetics and breeding of sorghum. The first phase of this project consisted of constructing a genetic map based on restriction fragment length polymorphisms (RFLPs). The ISU sorghum map was created through linkage analysis of 78 F2 plants of an intraspecific cross between inbred CK60 and accession PI229828. Subsequent mapping, efforts in several labs have enriched the sorghum map to the point where it now contains over 1,500 loci defined by RFLPs and many others defined by mutant phenotypes and QTLs. The ISU map consists of 201 loci distributed among 10 linkage groups covering 1299 cM. Comparison of sorghum and maize RFLP maps on the basis of common sets of DNA probes revealed a high degree of conservation as reflected by homology, copy number, and colinearity. Examples of conserved and rearranged locus orders were observed. The same sorghum population was used to map genetic factors (mutants and QTLS) for several traits including vegetative and reproductive morphology, maturity, insect, and disease resistance. Four QTLs for plant height, an important character for sorghum adaptation in temperate latitudes for grain production, were identified in a sample of 152 F2 plants whereas 6 QTLs were detected among their F3 progeny. These observations and assessments of other traits at 4 QTLs common to F2 plants and their F3 progeny indicate some of these regions correspond to loci (dw) previously identified on the basis of alleles with highly qualitative effects. Four of the six sorghum plant height QTLs seem to be orthologous to plant height QTLs in maize. Other possible instances of orthologous QTLs included regions for maturity and tillering. These observations suggest that the conservation of the maize and sorghum genomes encompasses sequence homology

  11. Sorghum to Ethanol Research

    Energy Technology Data Exchange (ETDEWEB)

    Dahlberg, Jeffrey A. [Univ. of California, Parlier, CA (United States). Kearney Research and Extension Center; Wolfrum, Edward J. [National Renewable Energy Lab. (NREL), Golden, CO (United States). Process and Analytical Engineering Group

    2010-09-28

    The development of a robust source of renewable transportation fuel will require a large amount of biomass feedstocks. It is generally accepted that in addition to agricultural and forestry residues, we will need crops grown specifically for subsequent conversion into fuels. There has been a lot of research on several of these so-called "dedicated bioenergy crops" including switchgrass, miscanthus, sugarcane, and poplar. It is likely that all of these crops will end up playing a role as feedstocks, depending on local environmental and market conditions. Many different types of sorghum have been grown to produce syrup, grain, and animal feed for many years. It has several features that may make it as compelling as other crops mentioned above as a renewable, sustainable biomass feedstock; however, very little work has been done to investigate sorghum as a dedicated bioenergy crop. The goal of this project was to investigate the feasibility of using sorghum biomass to produce ethanol. The work performed included a detailed examination of the agronomics and composition of a large number of sorghum varieties, laboratory experiments to convert sorghum to ethanol, and economic and life-cycle analyses of the sorghum-to-ethanol process. This work showed that sorghum has a very wide range of composition, which depended on the specific sorghum cultivar as well as the growing conditions. The results of laboratory- and pilot-scale experiments indicated that a typical high-biomass sorghum variety performed very similarly to corn stover during the multi-step process required to convert biomass feedstocks to ethanol; yields of ethanol for sorghum were very similar to the corn stover used as a control in these experiments. Based on multi-year agronomic data and theoretical ethanol production, sorghum can achieve more than 1,300 gallons of ethanol per acre given the correct genetics and environment. In summary, sorghum may be a compelling dedicated bioenergy crop that could help

  12. Harnessing the sorghum genome sequence:development of a genome-wide microsattelite (SSR) resource for swift genetic mapping and map based cloning in sorghum

    Science.gov (United States)

    Sorghum is the second cereal crop to have a full genome completely sequenced (Nature (2009), 457:551). This achievement is widely recognized as a scientific milestone for grass genetics and genomics in general. However, the true worth of genetic information lies in translating the sequence informa...

  13. Cloning and expression of a tomato glutathione S- transferase (GST ...

    African Journals Online (AJOL)

    In this study, ShGSTU1 was cloned into plasmid pET-28a, efficiently expressed in Escherichia coli upon isopropyl-β-D-1-thiogalactopyronoside (IPTG) induction, purified with Ni2+ affinity chromatography and biochemically characterized. The results show that the optimal conditions for the expression of recombinant ...

  14. Inheritance of Resistance to Sorghum Shoot Fly, Atherigona soccata in Sorghum, Sorghum bicolor (L. Moench

    Directory of Open Access Journals (Sweden)

    Mohammed eRiyazaddin

    2016-04-01

    Full Text Available Host plant resistance is one of the major components to control sorghum shoot fly, Atherigona soccata. To understand the nature of gene action for inheritance of shoot fly resistance, we evaluated 10 parents, 45 F1’s and their reciprocals in replicated trials during the rainy and postrainy seasons. Genotypes ICSV 700, Phule Anuradha, ICSV 25019, PS 35805, IS 2123, IS 2146 and IS 18551 exhibited resistance to shoot fly damage across seasons. Crosses between susceptible parents were preferred for egg laying by the shoot fly females, resulting in a susceptible reaction. ICSV 700, ICSV 25019, PS 35805, IS 2123, IS 2146 and IS 18551 exhibited significant and negative general combining ability (gca effects for oviposition, deadheart incidence, and overall resistance score. The plant morphological traits associated with expression of resistance/ susceptibility to shoot fly damage such as leaf glossiness, plant vigor, and leafsheath pigmentation also showed significant gca effects by these genotypes, suggesting the potential for use as a selection criterion to breed for resistance to shoot fly, A. soccata. ICSV 700, Phule Anuradha, IS 2146 and IS 18551 with significant positive gca effects for trichome density can also be utilised in improving sorghums for shoot fly resistance. The parents involved in hybrids with negative specific combining ability (sca effects for shoot fly resistance traits can be used in developing sorghum hybrids with adaptation to postrainy season. The significant reciprocal effects of combining abilities for oviposition, leaf glossy score and trichome density suggested the influence of cytoplasmic factors in inheritance of shoot fly resistance. Higher values of variance due to sca (σ2s, dominance variance (σ2d, and lower predictability ratios than the variance due to gca (σ2g and additive variance (σ2a for shoot fly resistance traits indicated the predominance of dominance type of gene action, whereas trichome density, leaf

  15. Successful pod infections by Moniliophthora roreri result in differential Theobroma cacao gene expression depending on the clone's level of tolerance.

    Science.gov (United States)

    Ali, Shahin S; Melnick, Rachel L; Crozier, Jayne; Phillips-Mora, Wilberth; Strem, Mary D; Shao, Jonathan; Zhang, Dapeng; Sicher, Richard; Meinhardt, Lyndel; Bailey, Bryan A

    2014-09-01

    An understanding of the tolerance mechanisms of Theobroma cacao used against Moniliophthora roreri, the causal agent of frosty pod rot, is important for the generation of stable disease-tolerant clones. A comparative view was obtained of transcript populations of infected pods from two susceptible and two tolerant clones using RNA sequence (RNA-Seq) analysis. A total of 3009 transcripts showed differential expression among clones. KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analysis of differentially expressed genes indicated shifts in 152 different metabolic pathways between the tolerant and susceptible clones. Real-time quantitative reverse transcription polymerase chain reaction (real-time qRT-PCR) analyses of 36 genes verified the differential expression. Regression analysis validated a uniform progression in gene expression in association with infection levels and fungal loads in the susceptible clones. Expression patterns observed in the susceptible clones diverged in tolerant clones, with many genes showing higher expression at a low level of infection and fungal load. Principal coordinate analyses of real-time qRT-PCR data separated the gene expression patterns between susceptible and tolerant clones for pods showing malformation. Although some genes were constitutively differentially expressed between clones, most results suggested that defence responses were induced at low fungal load in the tolerant clones. Several elicitor-responsive genes were highly expressed in tolerant clones, suggesting rapid recognition of the pathogen and induction of defence genes. Expression patterns suggested that the jasmonic acid-ethylene- and/or salicylic acid-mediated defence pathways were activated in the tolerant clones, being enhanced by reduced brassinosteroid (BR) biosynthesis and catabolic inactivation of both BR and abscisic acids. Finally, several genes associated with hypersensitive response-like cell death were also induced in tolerant clones. © 2014

  16. A novel approach to sequence validating protein expression clones with automated decision making

    Directory of Open Access Journals (Sweden)

    Mohr Stephanie E

    2007-06-01

    Full Text Available Abstract Background Whereas the molecular assembly of protein expression clones is readily automated and routinely accomplished in high throughput, sequence verification of these clones is still largely performed manually, an arduous and time consuming process. The ultimate goal of validation is to determine if a given plasmid clone matches its reference sequence sufficiently to be "acceptable" for use in protein expression experiments. Given the accelerating increase in availability of tens of thousands of unverified clones, there is a strong demand for rapid, efficient and accurate software that automates clone validation. Results We have developed an Automated Clone Evaluation (ACE system – the first comprehensive, multi-platform, web-based plasmid sequence verification software package. ACE automates the clone verification process by defining each clone sequence as a list of multidimensional discrepancy objects, each describing a difference between the clone and its expected sequence including the resulting polypeptide consequences. To evaluate clones automatically, this list can be compared against user acceptance criteria that specify the allowable number of discrepancies of each type. This strategy allows users to re-evaluate the same set of clones against different acceptance criteria as needed for use in other experiments. ACE manages the entire sequence validation process including contig management, identifying and annotating discrepancies, determining if discrepancies correspond to polymorphisms and clone finishing. Designed to manage thousands of clones simultaneously, ACE maintains a relational database to store information about clones at various completion stages, project processing parameters and acceptance criteria. In a direct comparison, the automated analysis by ACE took less time and was more accurate than a manual analysis of a 93 gene clone set. Conclusion ACE was designed to facilitate high throughput clone sequence

  17. Cloning, expression and purification of cold adapted acetate kinase ...

    African Journals Online (AJOL)

    shell) Neobuccinum living in the Antarctic ice-covered sea. An open reading frame of 1203 bp, coding for acetate kinase gene, called ack, was amplified, cloned into the expression vector, pETY-16b, and the enzyme was overproduced by ...

  18. Cloning, expression and functional analysis of MAP30 from ...

    African Journals Online (AJOL)

    use

    2011-12-07

    Dec 7, 2011 ... gene was cloned and expressed and the induction of the recombinant MAP30 protein on .... RNA reverse transcription was carried out by RevertAidTM First ... volume of Premix Ex Taq™ (Takara Bio Inc, Japan), PCR cycling.

  19. The diversity of local sorghum (Sorghum bicolor L. Moench) in Nusa Tenggara Timur province

    Science.gov (United States)

    Mukkun, L.; Lalel, H. J. D.; Richana, N.; Pabendon, M. B.; Kleden, S. R.

    2018-04-01

    Sorghum (Sorghum bicolor L. Moench) is an important food crop in the dry land including Nusa Tenggara Timur (NTT) Province. This plant has a high adaptability to drought, can produce on marginal land, and is relatively resistant to pests and diseases. The study aims to collect and identify the species of local sorghum being cultivated by farmers, and the purposes of cultivation. In addition, this study will preserve germ plasm of local sorghum by providing bank seeds for the next growing season. A collection of local sorghum samples was conducted in 7 districts using survey and observation method. A total of 53 species of sorghum were collected, with various characteristics and different local names. Based on the skin color of the seeds, the accessions were grouped into white groups (26.42%), light yellow (15.09%), black (20.75%), brown (24.52%), and red (13.20 %). Sorghum is used for complementary food for rice, consumption in times of food insecurity, fodder, and as a fence for corn and rice. It is necessary to characterize the type of local sorghum that has the potential to be developed for food, industrial raw materials, and for functional food.

  20. Consortium of eucalyptus with forage sorghum in semiarid of Minas Gerais State

    Directory of Open Access Journals (Sweden)

    Carlos Juliano Brant Albuquerque

    2017-11-01

    Full Text Available ABSTRACT: The objective was to estimate the wood yield and essential oil content in three clones of eucalyptus that were planted in four contrasting arrangements and intercropped with sorghum. Eucalyptus clones MA2001 (Eucalyptus camaldulensis x E. tereticornis, A144 (Eucalyptus urophylla x E. grandis, and GG100 (Eucalyptus urophylla x E. grandis, were planted in single rows (10x2m, double rows (2x3+15m and 2x3+20m; and, triple rows (2x3x2+10m in a randomized, complete block design experiment with four replicates. Our results demonstrated that planting spacing did not influence the essential oil yield or diameter at breast height in the clones. However, higher density plantings were shown to result in higher fresh weight of branches and leaves per plant. MA2001 grew taller, produced higher quantity of fresh biomass of branches and leaves per plant and volume of wood per hectare, and yielded more essential oil yield than the other clones. We concluded that MA2001 is the most suitable of the clones tested here for cultivation in water deficit conditions.

  1. In planta transformation of sorghum (Sorghum bicolor (L.) Moench)

    Indian Academy of Sciences (India)

    An in planta transformation protocol for sorghum (Sorghum bicolor (L.) Moench) using shoot apical meristem of germinating seedlings is reported in this study. Agrobacterium tumefaciens strain, LBA4404 with pCAMBIA1303 vector and construct pCAMBIA1303TPS1 were individually used for transformation. Since, the ...

  2. Biological activity evaluation of cloned and expressed caprine growth hormone from local Pakistani goat breed beetal

    International Nuclear Information System (INIS)

    Butt, H.I.; Shahzad, M.I.; Bashir, Q.

    2011-01-01

    Growth hormone cDNA of local goat breed-beetal was amplified by RT PCR and gene including leader sequence was cloned in pTZR57 cloning vector. The cGH-pTZR57 clone was confirmed by restriction digestion and sequence analyses before finally sub-cloning the gene in pND- a mammalian expression vector. The clones were again confirmed by restriction digestion and PCR analyses. Highly purified, supercoiled cGH-pND construct was used to transfect Vero cell lines for expression studies. The in vitro expression of cGH was checked by dot-ELISA technique. After confirming its in vitro cell line based expression, the construct was injected to 4 weeks old balb/c mice intramuscularly. Two animals were euthanized per week till four weeks to monitor the in vivo biological activity by evaluating the tibia epiphyseal width and body weight gain assays. Significant increase in tibia epiphyseal width and gain in body weight was observed from vaccinated animals. The study supports the concept that DNA based therapeutics are an efficient and cost effective method for gene delivery and in vivo transgene expressions. (author)

  3. In vitro binding of Sorghum bicolor transcription factors ABI4 and ABI5 to a conserved region of a GA 2-OXIDASE promoter: possible role of this interaction in the expression of seed dormancy

    OpenAIRE

    Cantoro, Renata; Crocco, Carlos Daniel; Benech-Arnold, Roberto Luis; Rodr?guez, Mar?a Ver?nica

    2013-01-01

    The precise adjustment of the timing of dormancy release according to final grain usage is still a challenge for many cereal crops. Grain sorghum [Sorghum bicolor (L.) Moench] shows wide intraspecific variability in dormancy level and susceptibility to pre-harvest sprouting (PHS). Both embryo sensitivity to abscisic acid (ABA) and gibberellin (GA) metabolism play an important role in the expression of dormancy of the developing sorghum grain. In previous works, it was shown that, simultaneous...

  4. Cloning, expression, purification and characterization of Leishmania tropica PDI-2 protein

    Directory of Open Access Journals (Sweden)

    Ali Dina

    2016-01-01

    Full Text Available In Leishmania species, protein disulfide isomerase (PDI is an essential enzyme that catalyzes thiol-disulfide interchange. The present work describes the isolation, cloning, sequencing and expression of the pdI-2 gene. Initially, the gene was amplified from L. tropica genomic DNA by PCR using specific primers before cloning into the expression vector pET-15b. The construct pET/pdI-2 was transformed into BL21(DE3 cells and induced for the protein expression. SDS-PAGE and western blot analysis showed that the expressed protein is about 51 kDa. Cloned gene sequence analysis revealed that the deduced amino acid sequence showed significant homology with those of several parasites PDIs. Finally, recombinant protein was purified with a metal-chelating affinity column. The putative protein was confirmed as a thiol - disulfide oxidoreductase by detecting its activity in an oxidoreductase assay. Assay result of assay suggested that the PDI-2 protein is required for both oxidation and reduction of disulfide bonds in vitro. Antibodies reactive with this 51 kDa protein were detected by Western blot analysis in sera from human infected with L. tropica. This work describes for the first time the enzymatic activity of recombinant L. tropica PDI-2 protein and suggests a role for this protein as an antigen for the detection of leishmaniasis infection.

  5. Genetic Analysis of Recombinant Inbred Lines for Sorghum bicolor ? Sorghum propinquum

    OpenAIRE

    Kong, Wenqian; Jin, Huizhe; Franks, Cleve D.; Kim, Changsoo; Bandopadhyay, Rajib; Rana, Mukesh K.; Auckland, Susan A.; Goff, Valorie H.; Rainville, Lisa K.; Burow, Gloria B.; Woodfin, Charles; Burke, John J.; Paterson, Andrew H.

    2013-01-01

    We describe a recombinant inbred line (RIL) population of 161 F5 genotypes for the widest euploid cross that can be made to cultivated sorghum (Sorghum bicolor) using conventional techniques, S. bicolor ? Sorghum propinquum, that segregates for many traits related to plant architecture, growth and development, reproduction, and life history. The genetic map of the S. bicolor ? S. propinquum RILs contains 141 loci on 10 linkage groups collectively spanning 773.1 cM. Although the genetic map ha...

  6. [Cloning of Clostridium perfringens alpha-toxin gene and extracellular expression in Escherichia coli].

    Science.gov (United States)

    Inoue, Masaharu; Kikuchi, Maho; Komoriya, Tomoe; Watanabe, Kunitomo; Kouno, Hideki

    2007-01-01

    Clostridium perfringens (C. perfringens) is a Gram-positive bacterial pathogen that widely propagets in the soil and the gastrointestinal tract of human and animals. This bacteria causes food poisoning, gas gangrene and other various range of infectious diseases. But there is no standard diagnosis method of C. perfringens. In order to develop a new type of immunoassay for clinical purpose, we studied expression and extracellular secretion of recombinant alpha-toxin having enzyme activity in E. coli expression system. Cloning was carried out after PCR amplification from C. perfringens GAI 94074 which was clinical isolate. Three kinds of fragment were cloned using pET100/D-TOPO vector. These fragments coded for ribosome binding site, signal peptide, and alpha-toxin gene respectively. Recombinant pET100 plasmid transformed into TOP 10 cells and the obtained plasmids were transformed into BL21 (DE3) cells. Then, the transformants were induced expression with IPTG. In conclusion, we successfully cloned, expressed and exteracellular secreted C. perfringens alpha-toxin containing signal peptide. Biologically, the obtained recombinant protein was positive for phospholipase C activity.

  7. Cloning, Expression and Purification of the Recombinant HIV-1 Tat-Nef Fusion Protein in Prokaryotic Expression System

    Directory of Open Access Journals (Sweden)

    Somayeh Kadkhodayan

    2016-07-01

    Full Text Available Abstract Background: Nef is one of the HIV-1 critical proteins, because it is essential for viral replication and AIDS disease progression and induction of immune response against it can partially inhibit viral infection. Moreover, a domain of the HIV-1 Trans-Activator of Transcription (Tat, 48-60 aa could act as a cell penetrating peptide (CPP. In current study, cloning and expression of Tat-Nef fusion protein was performed in E. coli for the first time. The protein expression was confirmed by western blot analysis and was purified using reverse staining method. Materials and Methods: In this experimental study, primarily, cloning of Tat-Nef fusion gene was done in pGEX6p2 expression vector. Then, the expression of Tat-Nef recombinat protein in E.coli BL21 (DE3 strain was performed by using IPTG inducer. The protein expression was confirmed by SDS-PAGE and western blotting using anti-Nef monoclonal antibody. Then, the recombinant fusion protein was purified from gel using reverse staining method. Results: The results of PCR analysis and enzyme digestion showed a clear band of ~ 726 bp in agarose gel indicating the correct Tat-Nef fusion cloning in pGEX6p2 prokaryotic expression vector. In addition, a 54 kDa band of Tat-Nef on SDS-PAGE revealed Tat-Nef protein expression that western blot analysis using anti-Nef monoclonal antibody confirmed it. Conclusion: The purified Tat-Nef recombinant fusion protein will be used as an antigen for protein vaccine design against HIV infection.

  8. Effects of the genotype and environment interaction on sugar accumulation in sweet sorghum varieties (Sorghum bicolor -{L.}- Moench grown in the lowland tropics of Colombia

    Directory of Open Access Journals (Sweden)

    Jaime Humberto Bernal

    2014-12-01

    Full Text Available Sugar production in sweet sorghums is affected by the environment. Therefore, in this study on the effects of the genotype x environment interaction on sugar accumulation, plant traits associated with the sugar content in the stem were evaluated in ten sorghum genotypes grown in six contrasting environments. The results indicated that the stem dry weight, juice sugar concentration (°Brix, stem sugar content and juice volume were controlled by the genetic constitution of the genotype, with a large environmental contribution to their expression. The results allowed for the identification of the sweet sorghum genotypes that have a high potential for the biofuel agroindustry due to their high sugar contents in the environmental conditions of Palmira, Espinal, Cerete and Codazzi. Humid tropical environments such as Gaitan and Villavicencio were less favorable for the competitive production of sweet sorghums for bioethanol due to their low levels of solar radiation and soil fertility.

  9. Cloning and functional expression of a human pancreatic islet glucose-transporter cDNA

    International Nuclear Information System (INIS)

    Permutt, M.A.; Koranyi, L.; Keller, K.; Lacy, P.E.; Scharp, D.W.; Mueckler, M.

    1989-01-01

    Previous studies have suggested that pancreatic islet glucose transport is mediated by a high-K m , low-affinity facilitated transporter similar to that expressed in liver. To determine the relationship between islet and liver glucose transporters, liver-type glucose-transporter cDNA clones were isolated from a human liver cDNA library. The liver-type glucose-transporter cDNA clone hybridized to mRNA transcripts of the same size in human liver and pancreatic islet RNA. A cDNA library was prepared from purified human pancreatic islet tissue and screened with human liver-type glucose-transporter cDNA. The authors isolated two overlapping cDNA clones encompassing 2600 base pairs, which encode a pancreatic islet protein identical in sequence to that of the putative liver-type glucose-transporter protein. Xenopus oocytes injected with synthetic mRNA transcribed from a full-length cDNA construct exhibited increased uptake of 2-deoxyglucose, confirming the functional identity of the clone. These cDNA clones can now be used to study regulation of expression of the gene and to assess the role of inherited defects in this gene as a candidate for inherited susceptibility to non-insulin-dependent diabetes mellitus

  10. Cloning and mRNA expression pattern analysis under low ...

    African Journals Online (AJOL)

    This research cloned endochitinase-antifreeze protein precursor (EAPP) gene of Dong-mu 70 rye (Secale cereale) by designing special primers according to Genbank's EAPP gene sequence, and analyzing the influence of low temperature stress on the expression of mRNA with RT-PCR. The results indicated that the ...

  11. a survey of sorghum downy mildew in sorghum in the sudano

    African Journals Online (AJOL)

    DR. AMINU

    Sahel savanna AEZs respectively) indicated that the disease was present only at the seedling stage ... In the southern guinea ... northern Nigeria, sorghum downy mildew in sorghum .... There was a significant (P>0.05) difference in SDM.

  12. Cloning of radiation-induced new gene RS1 expressed in mouse intestinal epithelium by enhanced RACE

    International Nuclear Information System (INIS)

    Wang Fengchao; Wang Junping; Su Yongping; Gao Jinsheng; Lou Shufen; Liu Xiaohong; Ren Jiong; Zhang Bo

    2003-01-01

    Objective: To obtain full-length cDNA of radiation-induced new gene RS1 expressed in mouse intestinal epithelium. Methods: The tissue expression profile of RS1 was analyzed by semi-quantitative RT-PCR to find the target tissue which highly expresses RS1. The total RNA extracted from the corresponding tissue was taken as the template for reverse-transcription. Enhanced RACE PCR was used to clone the full-length cDNA of RS1, including enrichment of the target gene through biotin-labeled probe for magnetic bead purification and nested PCR. Results: About a 2 kb long 3' end was successfully cloned and cloning of the 5' end proceeded well. Conclusion: The result is consistent with our experiment design. The set of combined techniques has been identified with the cloning of full-length cDNA from EST sequence especially when the optimal gene-specific primers are not available or the expression level of target gene is low

  13. Lipid transfer proteins from fruit: cloning, expression and quantification

    NARCIS (Netherlands)

    Zuidmeer, Laurian; van Leeuwen, W. Astrid; Budde, Ilona Kleine; Cornelissen, Jessica; Bulder, Ingrid; Rafalska, Ilona; Besolí, Noèlia Telléz; Akkerdaas, Jaap H.; Asero, Riccardo; Fernandez Rivas, Montserrat; Rivas, Montserrat Fernandez; Gonzalez Mancebo, Eloina; Mancebo, Eloina Gonzalez; van Ree, Ronald

    2005-01-01

    BACKGROUND: Lipid transfer proteins (LTP) are stable, potentially life-threatening allergens in fruits and many other vegetable foods. The aim of this study was to clone and express recombinant apple LTP (Mal d 3), as has previously been done for peach LTP (Pru p 3) and set up quantitative tests for

  14. TECHNOLOGICAL ADVANCES IN THE OBTAINING OF ETHANOL FROM Sweet sorghum (Sorghum bicolor (L. Moench

    Directory of Open Access Journals (Sweden)

    Sandro Pedroso Cunha

    2010-11-01

    Full Text Available ABSTRACT: Replacing the use of gasoline with ethanol in vehicles reduces by 90% CO2 emissions, this justifies the interest in the use of bioethanol as renewable energy. Besides sugar cane, cassava, maize and sugar beet special emphasis is being given to sorghum (Sorghum bicolor L. Moench to produce ethanol for its productivity and resistance. The sorghum is grown in Rio Grande do Sul with a production of about 70,000 tons / year. Embrapa has a program to develop cultivars of sorghum from the time the Pro-Alcohol and currently 25 new varieties of sorghum are being evaluated. Several factors are relevant in the optimization of production such as increased productivity and reduced costs in the production of ethanol. This study aimed to survey recent data that will assess production parameters of ethanol from sorghum. Factors such as reducing the risk of bacterial contamination, the means conducive to fermentation processes or grain sorghum stalk through the use of pretreatment of the sample, have been of great importance because it is basically turning cellulosic biomass into fermentable sugars. Superior genotypes of sweet sorghum for ethanol production are of utmost importance, as well as better ways to convert sugars into ethanol. Lignin, toxic against microorganisms, prevents the conversion of lignocellulose into ethanol. The conversion of lignocellulosic ethanol compounds based on the hydrolysis of cellulose producing simple sugars and fermenting those sugars into ethanol through microbiology.

  15. Influence of cloning by chromatin transfer on placental gene expression at Day 45 of pregnancy in cattle.

    Science.gov (United States)

    Mesquita, Fernando S; Machado, Sergio A; Drnevich, Jenny; Borowicz, Pawel; Wang, Zhongde; Nowak, Romana A

    2013-01-30

    Poor success rates in somatic cell cloning are often attributed to abnormal early embryonic development as well as late abnormal fetal growth and placental development. Although promising results have been reported following chromatin transfer (CT), a novel cloning method that includes the remodeling of the donor nuclei in vitro prior to their transfer into enucleated oocytes, animals cloned by CT show placental abnormalities similar to those observed following conventional nuclear transfer. We hypothesized that the placental gene expression pattern from cloned fetuses was ontologically related to the frequently observed placental phenotype. The aim of the present study was to compare global gene expression by microarray analysis of Day 44-47 cattle placentas derived from CT cloned fetuses with those derived from in vitro fertilization (i.e. control), and confirm the altered mRNA and protein expression of selected molecules by qRT-PCR and immunohistochemistry, respectively. The differentially expressed genes identified in the present study are known to be involved in a range of activities associated with cell adhesion, cell cycle control, intracellular transport and proteolysis. Specifically, an imprinted gene, involved with cell proliferation and placentomegaly in humans (CDKN1C) and a peptidase that serves as a marker for non-invasive trophoblast cells in human placentas (DPP4), had mRNA and protein altered in CT placentas. It was concluded that the altered pattern of gene expression observed in CT samples may contribute to the abnormal placental development phenotypes commonly identified in cloned offspring, and that expression of imprinted as well as trophoblast invasiveness-related genes is altered in cattle cloned by CT. Copyright © 2012 Elsevier B.V. All rights reserved.

  16. Protein expression of Myt272-3 recombinant clone and in silico ...

    African Journals Online (AJOL)

    Purpose: To investigate the expression of Myt272-3 recombinant protein and also to predict a possible protein vaccine candidate against Mycobacterium tuberculosis. Methods: Myt272-3 protein was expressed in pET30a+-Myt272-3 clone. The purity of the protein was determined using Dynabeads® His-Tag Isolation ...

  17. Cloning and expression of a widely expressed receptor tyrosine phosphatase

    DEFF Research Database (Denmark)

    Sap, J; D'Eustachio, P; Givol, D

    1990-01-01

    We describe the identification of a widely expressed receptor-type (transmembrane) protein tyrosine phosphatase (PTPase; EC 3.1.3.48). Screening of a mouse brain cDNA library under low-stringency conditions with a probe encompassing the intracellular (phosphatase) domain of the CD45 lymphocyte...... antigen yielded cDNA clones coding for a 794-amino acid transmembrane protein [hereafter referred to as receptor protein tyrosine phosphatase alpha (R-PTP-alpha)] with an intracellular domain displaying clear homology to the catalytic domains of CD45 and LAR (45% and 53%, respectively). The 142-amino acid...

  18. An integrated and comparative approach towards identification, characterization and functional annotation of candidate genes for drought tolerance in sorghum (Sorghum bicolor (L.) Moench).

    Science.gov (United States)

    Woldesemayat, Adugna Abdi; Van Heusden, Peter; Ndimba, Bongani K; Christoffels, Alan

    2017-12-22

    Drought is the most disastrous abiotic stress that severely affects agricultural productivity worldwide. Understanding the biological basis of drought-regulated traits, requires identification and an in-depth characterization of genetic determinants using model organisms and high-throughput technologies. However, studies on drought tolerance have generally been limited to traditional candidate gene approach that targets only a single gene in a pathway that is related to a trait. In this study, we used sorghum, one of the model crops that is well adapted to arid regions, to mine genes and define determinants for drought tolerance using drought expression libraries and RNA-seq data. We provide an integrated and comparative in silico candidate gene identification, characterization and annotation approach, with an emphasis on genes playing a prominent role in conferring drought tolerance in sorghum. A total of 470 non-redundant functionally annotated drought responsive genes (DRGs) were identified using experimental data from drought responses by employing pairwise sequence similarity searches, pathway and interpro-domain analysis, expression profiling and orthology relation. Comparison of the genomic locations between these genes and sorghum quantitative trait loci (QTLs) showed that 40% of these genes were co-localized with QTLs known for drought tolerance. The genome reannotation conducted using the Program to Assemble Spliced Alignment (PASA), resulted in 9.6% of existing single gene models being updated. In addition, 210 putative novel genes were identified using AUGUSTUS and PASA based analysis on expression dataset. Among these, 50% were single exonic, 69.5% represented drought responsive and 5.7% were complete gene structure models. Analysis of biochemical metabolism revealed 14 metabolic pathways that are related to drought tolerance and also had a strong biological network, among categories of genes involved. Identification of these pathways, signifies the

  19. Cloning and heterologous expression of a gene encoding lycopene ...

    African Journals Online (AJOL)

    This report describes the cloning and expression of a gene lycopene epsilon cyclase, (LCYE) from Camellia sinensis var assamica which is a precursor of the carotenoid lutein in tea. The 1982 bp cDNA sequence with 1599 bp open reading frame of LCYE was identified from an SSH library constructed for quality trait in tea.

  20. Use of hybridization (F1 in forage sorghum (Sorghum bicolor (L. Moench breeding

    Directory of Open Access Journals (Sweden)

    Pataki Imre

    2010-01-01

    Full Text Available In plants with bisexual flowers, the development of hybrids and F1 seed production is only possible by using cytoplasmatic male sterility. The discovery of such sterility and the maintainers has made it possible to utilize the phenomenon of heterosis to improve yields and yield components in forage sorghum. It has been shown that the best way to develop forage sorghum hybrids is to cross grain sorghum as the female parent and Sudan grass as the male. The objective of this study was to develop a forage sorghum hybrid for the production of green matter to be used either fresh or for silage. The sorghum hybrid developed in these efforts (Siloking is intended for multiple cutting, as the basal nodes produce buds and regrowth takes place. The performance of the new hybrid with respect to yield and quality was compared to that of the forage sorghum cultivar NS Džin. In a two-year study conducted under different growing conditions in four locations, Siloking produced an average green matter yield of 86.29 t ha-1 (two cuts, a dry matter yield of 25.34 t ha-1, and a crude protein content of 11.85 %. Siloking outperformed NS Džin in terms of yield and quality. .

  1. An array of Escherichia coli clones over-expressing essential proteins: A new strategy of identifying cellular targets of potent antibacterial compounds

    International Nuclear Information System (INIS)

    Xu, H. Howard; Real, Lilian; Bailey, Melissa Wu

    2006-01-01

    With the advancement of high throughput screening, it has become easier and faster to discover hit compounds that inhibit proliferation of bacterial cells. However, development in technologies used to identify cellular targets of potent antibacterial inhibitors has lagged behind. Here, we describe a novel strategy of target identification for antibacterial inhibitors using an array of Escherichia coli clones each over-expressing one essential protein. In a proof-of-concept study, eight essential genes were cloned into pLex5BA vector under the control of an inducible promoter. Over-expression of target proteins was confirmed. For two clones, one over-expressing FabI and the other over-expressing MurA enzymes, the host cells became 17- and 139-fold more resistant to the specific inhibitors triclosan and phosphomycin, respectively, while the susceptibility of other clones towards these inhibitors remained unchanged after induction of gene expression. Target identification via target protein over-expression was demonstrated using both mixed clone and individual clone assay formats

  2. (cucurbita pepo) and sorghum

    African Journals Online (AJOL)

    big timmy

    ... properties of pumpkin (Cucurbita pepo) and sorghum (Sorghum bicolor) flour blends fermented with pure strains of Lactobacillus ... good storage characteristics and affordable cost. (Akinrele ... (MRS), Nutrient agar (NA) and Potato dextrose.

  3. Transcriptome Characterization and Functional Marker Development in Sorghum Sudanense.

    Directory of Open Access Journals (Sweden)

    Jieqin Li

    Full Text Available Sudangrass, Sorghum sudanense, is an important forage in warm regions. But little is known about its genome. In this study, the transcriptomes of sudangrass S722 and sorghum Tx623B were sequenced by Illumina sequencing. More than 4Gb bases were sequenced for each library. For Tx623B and S722, 88.79% and 83.88% reads, respectively were matched to the Sorghum bicolor genome. A total of 2,397 differentially expressed genes (DEGs were detected by RNA-Seq between the two libraries, including 849 up-regulated genes and 1,548 down-regulated genes. These DEGs could be divided into three groups by annotation analysis. A total of 44,495 single nucleotide polymorphisms (SNPs were discovered by aligning S722 reads to the sorghum reference genome. Of these SNPs, 61.37% were transition, and this value did not differ much between different chromosomes. In addition, 16,928 insertion and deletion (indel loci were identified between the two genomes. A total of 5,344 indel markers were designed, 15 of which were selected to construct the genetic map derived from the cross of Tx623A and Sa. It was indicated that the indel markers were useful and versatile between sorghum and sudangrass. Comparison of synonymous base substitutions (Ks and non-synonymous base substitutions (Ka between the two libraries showed that 95% orthologous pairs exhibited Ka/Ks<1.0, indicating that these genes were influenced by purifying selection. The results from this study provide important information for molecular genetic research and a rich resource for marker development in sudangrass and other Sorghum species.

  4. Sorghums: viable biomass candidates

    Energy Technology Data Exchange (ETDEWEB)

    McClure, T A; Arthur, M F; Kresovich, S; Scantland, D A

    1980-01-01

    Agronomic studies conducted at Battelle's Columbus Division to evaluate biomass and sugar yields of sweet sorghum are described and the major findings are summarized. Development opportunities for using sorghum cultivars as a large-scale energy crop are discussed. With presently available cultivars, sweet sorghum should produce 3500 to 4000 liters ethanol per hectare from the fermentable sugars alone. Conversion of the stalk fibers into alcohol could increase production by another 1600 to 1900 liters per hectare with existing cultivars. These yields are approximately 30 to 40% greater per hectare than would be obtained from above average yields of grain and stalk fiber with corn. There is reason to believe, that with hybrid sweet sorghum, these yields could be further increased by as much as 30%. Diminishing land availability for agricultural crops necessitates that maximum yields be obtained. Over the next decade, imaginative technological innovations in sorghum harvesting, processing, and crop preservation, coupled with plant breeding research should help this crop realize its full potential as a renewable resource for energy production.

  5. Identification and characterization of a novel gene differentially expressed in zebrafish cross-subfamily cloned embryos

    Directory of Open Access Journals (Sweden)

    Wang Ya-Ping

    2008-03-01

    Full Text Available Abstract Background Cross-species nuclear transfer has been shown to be a potent approach to retain the genetic viability of a certain species near extinction. However, most embryos produced by cross-species nuclear transfer were compromised because that they were unable to develop to later stages. Gene expression analysis of cross-species cloned embryos will yield new insights into the regulatory mechanisms involved in cross-species nuclear transfer and embryonic development. Results A novel gene, K31, was identified as an up-regulated gene in fish cross-subfamily cloned embryos using SSH approach and RACE method. K31 complete cDNA sequence is 1106 base pairs (bp in length, with a 342 bp open reading frame (ORF encoding a putative protein of 113 amino acids (aa. Comparative analysis revealed no homologous known gene in zebrafish and other species database. K31 protein contains a putative transmembrane helix and five putative phosphorylation sites but without a signal peptide. Expression pattern analysis by real time RT-PCR and whole-mount in situ hybridization (WISH shows that it has the characteristics of constitutively expressed gene. Sub-cellular localization assay shows that K31 protein can not penetrate the nuclei. Interestingly, over-expression of K31 gene can cause lethality in the epithelioma papulosum cyprinid (EPC cells in cell culture, which gave hint to the inefficient reprogramming events occurred in cloned embryos. Conclusion Taken together, our findings indicated that K31 gene is a novel gene differentially expressed in fish cross-subfamily cloned embryos and over-expression of K31 gene can cause lethality of cultured fish cells. To our knowledge, this is the first report on the determination of novel genes involved in nucleo-cytoplasmic interaction of fish cross-subfamily cloned embryos.

  6. Identification of cDNA clones expressing immunodiagnostic antigens from Trichinella spiralis

    International Nuclear Information System (INIS)

    Zarlenga, D.; Gamble, H.R.

    1987-01-01

    A cDNA expression library was built in lambda gt11 phage using poly A mRNA isolated from Trichinella spiralis muscle stage larvae. This library was screened with rabbit antibodies to parasite excretory-secretory (ES) products and greater than 180 clones were isolated. Thirteen clones producing highly immunogenic protein antigens were plaque purified and rescreened with pig antisera to T.spiralis, Trichuris suis or Ascaris suum to identify clones producing epitopes specific to T.spiralis ES products, only. Two clones, TsAc-2 and TsAc-8, which displayed strong interactions with pig antisera to T. spiralis were lysogenized in E. coli Y1089 and the protein extracted. Western blots of the crude fusion proteins revealed molecular weights of 133 kD and 129 kD, respectively. Northern blot analysis of total RNA with 32 P labelled cDNA:lambda gt11 probes indicated single RNA transcripts for each clone with molecular sizes corresponding to 800-850 nucleotides. dscDNA inserts were estimated by southern blot analysis to be 500 bp and 340 bp, respectively, with no cross-hybridization observed between the cloned sequences. Dot blots using pig sera to screen crude fusion protein preparations, total bacterial protein (negative controls) and crude worm extract or ES products from T.spiralis, T.suis and A.suum (positive controls) corroborated the specificity and sensitivity of these clones as potential diagnostic antigens for swine trichinellosis

  7. [Cloning and characterization of genes differentially expressed in human dental pulp cells and gingival fibroblasts].

    Science.gov (United States)

    Wang, Zhong-dong; Wu, Ji-nan; Zhou, Lin; Ling, Jun-qi; Guo, Xi-min; Xiao, Ming-zhen; Zhu, Feng; Pu, Qin; Chai, Yu-bo; Zhao, Zhong-liang

    2007-02-01

    To study the biological properties of human dental pulp cells (HDPC) by cloning and analysis of genes differentially expressed in HDPC in comparison with human gingival fibroblasts (HGF). HDPC and HGF were cultured and identified by immunocytochemistry. HPDC and HGF subtractive cDNA library was established by PCR-based modified subtractive hybridization, genes differentially expressed by HPDC were cloned, sequenced and compared to find homogeneous sequence in GenBank by BLAST. Cloning and sequencing analysis indicate 12 genes differentially expressed were obtained, in which two were unknown genes. Among the 10 known genes, 4 were related to signal transduction, 2 were related to trans-membrane transportation (both cell membrane and nuclear membrane), and 2 were related to RNA splicing mechanisms. The biological properties of HPDC are determined by the differential expression of some genes and the growth and differentiation of HPDC are associated to the dynamic protein synthesis and secretion activities of the cell.

  8. Cloning and expression of the Legionella micdadei "common antigen" in Escherichia coli

    DEFF Research Database (Denmark)

    Bangsborg, Jette Marie; Collins, M T; Høiby, N

    1989-01-01

    To study individual Legionella antigens, a Legionella micdadei genomic library in Escherichia coli SC181 was established. Partially Sau3A digested L. micdadei DNA fragments (15-25 kilobase pairs (kb] were cloned into the tetracycline resistance gene of the cosmid vector pHC79. Four thousand...... ampicillin resistant recombinants were obtained; seven hundred were screened for expression of Legionella antigens in Western blot analysis with a polyspecific E. coli-absorbed anti-L. micdadei rabbit antibody. One of the positive clones expressed a 60 kilodalton (K) antigen, which reacted strongly...... will provide important information with respect to genetic vs. antigenic relatedness among Legionellae and other Gram-negative species, as well as to CA structure and possible function....

  9. Analysis of aluminium sensitivity in sorghum (Sorghum bicolor (L.) Moench) genotypes

    NARCIS (Netherlands)

    Tan, K.

    1993-01-01

    Twelve genotypes of sorghum ( Sorghum bicolor (L.) Moench) differing in Al sensitivity were grown in an acid soil (with additions of lime or MgSO 4 ) and in nutrient solutions (with or without Al at constant pH) for periods between 14 and 35 days.

  10. Genetic Dissection of Bioenergy-Related Traits in Sweet Sorghum (Sorghum bicolor) under Danish Agro-Climatic Conditions

    DEFF Research Database (Denmark)

    Mocoeur, Anne Raymonde Joelle

    Sorghum (Sorghum bicolor (L.) Moench), a C4 African originated grass, ranks 5th most important crop worldwide, feeding over 500 million people in tropical regions as it withstands a wide panel of biotic and abiotic stresses. The small and simple diploid genome of sorghum was elected as the third...... plant for sequencing in 2009 promoting it as a C4 model plant. Among the very diverse genetic resources available for sorghum, sweet sorghum plants; amassing large quantities of juice-rich and sugar-rich stem, grain and vegetative biomass; have been enlightened as bioenergy crop as it can produced from...... a single plant food, feed and fuel. Sweet sorghum has gained interest in Europe to replace maize, for biogas and bioenergy productions, but this versatile crop is sensitive to chilling temperatures and little breeding efforts have been done toward its cold acclimation. The state-of-art of using...

  11. Cloning and Expression of Luteinizing Hormone Subunits in Chinese Hamster Ovary Cell Line

    Directory of Open Access Journals (Sweden)

    Zeinab Soleimanifar

    2016-10-01

    Full Text Available Background: Luteinizing hormone (LH was secreted by the stimulating cells of the testes and ovaries in the anterior pituitary gland. The application of this hormone is in the treatment of men and women with infertility and amenorrhea respectively.Materials and Methods: In the present study the alpha and beta subunits of human LH gene were cloned into the pEGFP-N1 expression vector and produced the recombinant LH hormone in Chinese hamster ovary (CHO eukaryotic system.Results: Alpha and beta subunits of LH hormone were cloned between NheI and BamHI cut sites of pEGFP_N1 expression plasmid and confirmed by PCR.  Hormone expression was evaluated in CHO cell line by Western blotting using the specific antibody.Conclusion: Alpha and beta subunits of LH hormone were expressed in CHO cell line perfectly.

  12. Microsatellite markers reveal a predominant sugarcane aphid (Homoptera: Aphididae) clone is found on sorghum in seven states and one territory of the USA

    Science.gov (United States)

    The sugarcane aphid, Melanaphis sacchari, has become a serious pest causing severe economic losses to sorghum grown in the southern United States (U.S.). Since its original detection in four states in 2013, M. sacchari on sorghum has now, for 2016, spread to 19 states. The presence of one or multip...

  13. Real-Time Determination of Photosynthesis, Transpiration, Water-Use Efficiency and Gene Expression of Two Sorghum bicolor (Moench Genotypes Subjected to Dry-Down

    Directory of Open Access Journals (Sweden)

    Alessandra Fracasso

    2017-05-01

    Full Text Available Plant growth and productivity are strongly affected by limited water availability in drought prone environments. The current climate change scenario, characterized by long periods without precipitations followed by short but intense rainfall, forces plants to implement different strategies to cope with drought stress. Understanding how plants use water during periods of limited water availability is of primary importance to identify and select the best adapted genotypes to a certain environment. Two sorghum genotypes IS22330 and IS20351, previously characterized as drought tolerant and drought sensitive genotypes, were subjected to progressive drought stress through a dry-down experiment. A whole-canopy multi-chamber system was used to determine the in vivo water use efficiency (WUE. This system records whole-canopy net photosynthetic and transpiration rate of 12 chambers five times per hour allowing the calculation of whole-canopy instantaneous WUE daily trends. Daily net photosynthesis and transpiration rates were coupled with gene expression dynamics of five drought related genes. Under drought stress, the tolerant genotype increased expression level for all the genes analyzed, whilst the opposite trend was highlighted by the drought sensitive genotype. Correlation between gene expression dynamics and gas exchange measurements allowed to identify three genes as valuable candidate to assess drought tolerance in sorghum.

  14. Real-Time Determination of Photosynthesis, Transpiration, Water-Use Efficiency and Gene Expression of Two Sorghum bicolor (Moench) Genotypes Subjected to Dry-Down.

    Science.gov (United States)

    Fracasso, Alessandra; Magnanini, Eugenio; Marocco, Adriano; Amaducci, Stefano

    2017-01-01

    Plant growth and productivity are strongly affected by limited water availability in drought prone environments. The current climate change scenario, characterized by long periods without precipitations followed by short but intense rainfall, forces plants to implement different strategies to cope with drought stress. Understanding how plants use water during periods of limited water availability is of primary importance to identify and select the best adapted genotypes to a certain environment. Two sorghum genotypes IS22330 and IS20351, previously characterized as drought tolerant and drought sensitive genotypes, were subjected to progressive drought stress through a dry-down experiment. A whole-canopy multi-chamber system was used to determine the in vivo water use efficiency (WUE). This system records whole-canopy net photosynthetic and transpiration rate of 12 chambers five times per hour allowing the calculation of whole-canopy instantaneous WUE daily trends. Daily net photosynthesis and transpiration rates were coupled with gene expression dynamics of five drought related genes. Under drought stress, the tolerant genotype increased expression level for all the genes analyzed, whilst the opposite trend was highlighted by the drought sensitive genotype. Correlation between gene expression dynamics and gas exchange measurements allowed to identify three genes as valuable candidate to assess drought tolerance in sorghum.

  15. Human Interleukine-1 receptor antagonist:Cloning, Expression and Optimization in E.coli Host

    Directory of Open Access Journals (Sweden)

    Gh. Barati

    2014-07-01

    Full Text Available Introduction & Objective: Interleukine-1 receptor antagonist (IL-1RA is a powerful anti-inflammatory cytokine which limits the biological effects of IL-1. Due to structural similarity between IL-1 and its antagonist, IL-1RA competitively binds to IL-1 receptor which leads to no signal transduction. Therefore , it is applied in the treatment of patients with inflammatory diseases such as Rheumatoid Arthritis. The aim of this study is cloning, expression and op-timization of IL-1RA in E. coli. Materials & Methods: In this experimental study synthetically prepared cDNA was amplified by PCR. After double digestion with NdeI and XhoI restriction enzymes, this gene was cloned in pET28a expression vector. Expression of desired gene was analyzed at RNA level by RT-PCR and at protein level by SDS-PAGE and followed by western blot to confirm SDS-PAGE results. Optimization of recombinant protein expression was performed in dif-ferent IPTG concentrations and harvesting times after induction. Results: The presence of gene in pET28a was determined by colony-PCR and confirmed by restriction digestion. Transcription of cloned gene and expression of high yield recombinant protein were shown by RT-PCR and SDS-PAGE, respectively. The result of SDS-PAGE was confirmed by western blot. Expression was optimized in different induction time and IPTG concentrations Conclusion: The result of this study demonstrated expression of this recombinant protein at high level in E.coli system by pET28a expression vector. This study also showed a direct as-sociation between the increased level of expression and time of induction . Therefore, an overnight induction time with 0.1 mM IPTG concentration is recommended for a high level expression. (Sci J Hamadan Univ Med Sci 2014; 21 (2:145-151

  16. Energy analysis of ethanol production from sweet sorghum

    Energy Technology Data Exchange (ETDEWEB)

    Worley, J.W. (Georgia Univ., Athens, GA (United States). Dept. of Agricultural Engineering); Vaughan, D.H.; Cundiff, J.S. (Virginia Polytechnic Inst. and State Univ., Blacksburg, VA (United States). Dept. of Agricultural Engineering)

    1992-01-01

    The Piedmont System is a collection of equipment for efficiently removing the juice from sweet sorghum stalks for the production of ethanol. The concept is to separate the whole stalks into pith and rind-leaf fractions, pass only the pith fraction through a screw press, and thus achieve an improvement in juice-expression efficiency and press capacity. An energy analysis was done for two options of this proposed harvesting/processing system: (Option 1) The juice is evaporated to syrup and used throughout the year to produce ethanol, and the by-products are used as cattle feed. (Option 2) The juice is fermented as it is harvested, and the by-products (along with other cellulosic materials) are used as feedstock for the remainder of the year. Energy ratios (energy output/energy input) of 0.9, 1.1 and 0.8 were found for sweet sorghum Option 1, sweet sorghum Option 2, and corn, respectively, as feedstocks for ethanol. If only liquid fuels are considered, the ratios are increased to 3.5, 7.9 and 4.5. (author).

  17. Expression of Two N1 Clones with Single Amino Acid Dissimilarity of Avian Influenza H5N1 Virus

    Directory of Open Access Journals (Sweden)

    RISZA HARTAWAN

    2012-12-01

    Full Text Available Two clones of N1 gene derived from isolate A/Dk/Tangerang/Bbalitvet-ACIAR-TE11/2007 (H5N1 exhibit single mismatch of amino acid sequence at position 242 that is threonine and methionine for the clone #3 and #5, respectively. In order to evaluate the effect of the amino acid substitution, these clones were inserted into two different expression vectors that are pEGFP-C1 and pcDNA-3.3 TOPO® TA cloning. Subsequently, the respective recombinant clones were transfected into eukaryotic cells, including CEF, RK13 and VERO using Lipofectamine ‘plus’ reagent. As a result, the clone #3 retaining atypical sequence showed lower expression level rather than the clone #15 in both vectors and all type of cells. The 3D conformational modelling revealed that the mutation occurs in the inner part of glycoprotein embedded within envelope or matrix. Therefore, the missense mutation seems has no effect on the antigenic properties of neuraminidase but this substitution by any means causes lethal mutagenesis in the individual gene expression by reducing level of protein transcript.

  18. Cloning and expression of calmodulin gene in Scoparia dulcis.

    Science.gov (United States)

    Saitoh, Daisuke; Asakura, Yuki; Nkembo, Marguerite Kasidimoko; Shite, Masato; Sugiyama, Ryuji; Lee, Jung-Bum; Hayashi, Toshimitsu; Kurosaki, Fumiya

    2007-06-01

    A homology-based cloning strategy yielded a cDNA clone, designated Sd-cam, encoding calmodulin protein from Scoparia dulcis. The restriction digests of genomic DNA of S. dulcis showed a single hybridized signal when probed with the fragment of this gene in Southern blot analyses, suggesting that Sd-cam occurs as a sole gene encoding calmodulin in the plant. The reverse-transcription polymerase chain reaction analysis revealed that Sd-cam was appreciably expressed in leaf, root and stem tissues. It appeared that transcription of this gene increased transiently when the leaf cultures of S. dulcis were treated with methyl jasmonate and calcium ionophore A23187. These results suggest that transcriptional activation of Sd-cam is one of the early cellular events of the methyl jasmonate-induced responses of S. dulcis.

  19. Enhanced specificity in immunoscreening of expression cDNA clones using radiolabeled antigen overlay

    International Nuclear Information System (INIS)

    Chao, S.; Chao, L.; Chao, J.

    1989-01-01

    A highly sensitive and specific method has been developed for immunoscreening clones from an expression cDNA library. The procedures utilize a radiolabeled antigen detection method described originally for the immunoblotting of plasma proteins. Screening of rat alpha 1-antitrypsin clones was used. Comparison between Western blots of alpha 1-antitrypsin using both labeled antigen and protein A detection methods showed that the former yielded lower background and greater sensitivity than the latter. Further, this technique was shown to have a lower detection limit of less than 20 ng through Western blot analysis of varying concentrations of alpha 1-antitrypsin. The procedures are based on the expression of the protein by cDNA clones containing the DNA inserts in the correct reading frame. Following the transfer of phage proteins to nitrocellulose membranes, the bivalent antibodies bind monovalently to both nitrocellulose-bound-antigen in the phage lysates and radiolabeled antigen. The radiolabeled antigen overlay method is superior to the protein A detection method in sensitivity, specificity and reproducibility. This improved method can be applied in general for screening expression cDNA libraries, provided that the specific antiserum and radiolabeled antigen are available

  20. Cloning and over-expression of Penicillin G acylase in Escherichia ...

    African Journals Online (AJOL)

    The aim of this study is to screen for PGA producing Escherichia coli isolates as well as the cloning and recombinant expression of PGA for high level enzyme production. Bacteria isolated from environmental and clinical samples were identified by standard microbiological tests and then E. coli isolates were subjected to

  1. Molecular transformation, gene cloning, and gene expression systems for filamentous fungi

    Science.gov (United States)

    Gold, Scott E.; Duick, John W.; Redman, Regina S.; Rodriguez, Rusty J.

    2001-01-01

    This chapter discusses the molecular transformation, gene cloning, and gene expression systems for filamentous fungi. Molecular transformation involves the movement of discrete amounts of DNA into cells, the expression of genes on the transported DNA, and the sustainable replication of the transforming DNA. The ability to transform fungi is dependent on the stable replication and expression of genes located on the transforming DNA. Three phenomena observed in bacteria, that is, competence, plasmids, and restriction enzymes to facilitate cloning, were responsible for the development of molecular transformation in fungi. Initial transformation success with filamentous fungi, involving the complementation of auxotrophic mutants by exposure to sheared genomic DNA or RNA from wt isolates, occurred with low transformation efficiencies. In addition, it was difficult to retrieve complementing DNA fragments and isolate genes of interest. This prompted the development of transformation vectors and methods to increase efficiencies. The physiological studies performed with fungi indicated that the cell wall could be removed to generate protoplasts. It was evident that protoplasts could be transformed with significantly greater efficiencies than walled cells.

  2. Effect of emulsifiers on complexation and retrogradation characteristics of native and chemically modified White sorghum (Sorghum bicolor) starch

    International Nuclear Information System (INIS)

    Ali, Tahira Mohsin; Hasnain, Abid

    2013-01-01

    Highlights: ► Sorghum starches were chemically modified. ► Starch–lipid complexes were studied in the presence of emulsifiers. ► Type II complexes were also detected in native and oxidized starches on adding GMS. ► Starch–lipid complexes sharply reduced retrogradation in modified starches. - Abstract: The effect of emulsifiers on complexation and retrogradation characteristics of native and chemically modified white sorghum starches was studied. Complex forming tendency of white sorghum starch with commercially available emulsifiers GMS and DATEM improved after acetylation. Presence of emulsifiers reduced λ max (wavelength of maximum absorbance) both for native and modified sorghum starches suggesting lower availability of amylose chains to complex with iodine. In native white sorghum starch (NWSS) and oxidized white sorghum starch (OWSS), both Type I and Type II starch–lipid complexes were observed on addition of 1.0% GMS prior to gelatinization. Acetylated-oxidized white sorghum starch (AOWSS) formed weakest complexes among all the modified starches. The results revealed that antistaling characteristics of modified sorghum starches were enhanced when used in combination with emulsifiers. The most prominent decline in reassociative capability among modified starches was observed for acetylated starches.

  3. Clone and expression of human transferrin receptor gene: a marker gene for magnetic resonance imaging

    International Nuclear Information System (INIS)

    Li Li; Liu Lizhi; Lv Yanchun; Liu Xuewen; Cui Chunyan; Wu Peihong; Liu Qicai; Ou Shanxing

    2007-01-01

    Objective: To clone human transferrin receptor (hTfR) gene and construct expression vector producing recombination protein. Methods: Human transferrin receptor gene cDNA was amplified by RT-PCR from human embryonic liver and lung tissue. Recombinant pcDNA3-hTfR and pEGFP-Cl-hTfR plasmids were constructed and confirmed by DNA sequencing. These plasmids were stably transfected into the HEK293 cells. The protein expression in vitro was confirmed by Western Blot. The efficiency of expression and the location of hTfR were also investigated by fluorescence microscopy and confocal fluorescence microscopy. Results: The full length cDNA of hTfR gene (2332 bp) was cloned and sequenced. The hTfR (190 000) was overexpressed in transfected HEK293 cells by Western blot analysis. Fluorescence micrographs displayed that the hTfR was expressed at high level and located predominantly in the cell surface. Conclusions: Human transferrin receptor (hTfR) gene has been successfully cloned and obtained high-level expression in HEK293 cells, and the recombination protein of hTfR distributed predominantly in the cell membrane. (authors)

  4. Review of Sorghum Production Practices: Applications for Bioenergy

    Energy Technology Data Exchange (ETDEWEB)

    Turhollow Jr, Anthony F [ORNL; Webb, Erin [ORNL; Downing, Mark [ORNL

    2010-06-01

    Sorghum has great potential as an annual energy crop. While primarily grown for its grain, sorghum can also be grown for animal feed and sugar. Sorghum is morphologically diverse, with grain sorghum being of relatively short stature and grown for grain, while forage and sweet sorghums are tall and grown primarily for their biomass. Under water-limited conditions sorghum is reliably more productive than corn. While a relatively minor crop in the United States (about 2% of planted cropland), sorghum is important in Africa and parts of Asia. While sorghum is a relatively efficient user of water, it biomass potential is limited by available moisture. The following exhaustive literature review of sorghum production practices was developed by researchers at Oak Ridge National Laboratory to document the current state of knowledge regarding sorghum production and, based on this, suggest areas of research needed to develop sorghum as a commercial bioenergy feedstock. This work began as part of the China Biofuels Project sponsored by the DOE Energy Efficiency and Renewable Energy Program to communicate technical information regarding bioenergy feedstocks to government and industry partners in China, but will be utilized in a variety of programs in which evaluation of sorghum for bioenergy is needed. This report can also be used as a basis for data (yield, water use, etc.) for US and international bioenergy feedstock supply modeling efforts.

  5. Development of new USER-based cloning vectors for multiple genes expression in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Kildegaard, Kanchana Rueksomtawin; Jensen, Niels Bjerg; Maury, Jerome

    2013-01-01

    auxotrophic and dominant markers for convenience of use. Our vector set also contains both integrating and multicopy vectors for stability of protein expression and high expression level. We will make the new vector system available to the yeast community and provide a comprehensive protocol for cloning...... the production strain with the proper phenotype and product yield. However, the sequential number of metabolic engineering is time-consuming. Furthermore, the number of available selectable markers is also limiting the number of genetic modifications. To overcome these limitations, we have developed a new set...... of shuttle vectors for convenience of use for high-throughput cloning and selectable marker recycling. The new USER-based cloning vectors consist of a unique USER site and a CRE-loxP-mediated marker recycling system. The USER site allows insertion of genes of interest along with a bidirectional promoter...

  6. Radioinduced variation in genetic improvement of sorghum (Sorghum bicolor (l.). Moench)

    International Nuclear Information System (INIS)

    Gutierrez del Rio, E.

    1984-01-01

    A genetic variability study among 25 varieties of sorghum (Sorghum bicolor (L.) Moench) is presented. The populations are irradiated with 0, 10, 20, 30, 40, 50 and 60 Krads of cobalt 60 as far as M 5 generation. An individual selection is done taking into consideration agronomic characteristics like precocity, type, size. height of the plant. (M.A.C.) [pt

  7. Effect of Harvesting Stage on Sweet Sorghum (Sorghum bicolor L. Genotypes in Western Kenya

    Directory of Open Access Journals (Sweden)

    Moses Owuor Oyier

    2017-01-01

    Full Text Available Harvesting stage of sweet sorghum (Sorghum bicolor L. Moench cane is an important aspect in the content of sugar for production of industrial alcohol. Four sweet sorghum genotypes were evaluated for harvesting stage in a randomized complete block design. In order to determine sorghum harvest growth stage for bioethanol production, sorghum canes were harvested at intervals of seven days after anthesis. The genotypes were evaluated at different stages of development for maximum production of bioethanol from flowering to physiological maturity. The canes were crushed and juice fermented to produce ethanol. Measurements of chlorophyll were taken at various stages as well as panicles from the harvested canes. Dried kernels at 14% moisture content were also weighed at various stages. Chlorophyll, grain weight, absolute ethanol volume, juice volume, cane yield, and brix showed significant (p=0.05 differences for genotypes as well as the stages of harvesting. Results from this study showed that harvesting sweet sorghum at stages IV and V (104 to 117 days after planting would be appropriate for production of kernels and ethanol. EUSS10 has the highest ethanol potential (1062.78 l ha−1 due to excellent juice volume (22976.9 l ha−1 and EUSS11 (985.26 l ha−1 due to its high brix (16.21.

  8. Cloning, expression and characterization of alcohol dehydrogenases in the silkworm Bombyx mori

    Directory of Open Access Journals (Sweden)

    Nan Wang

    2011-01-01

    Full Text Available Alcohol dehydrogenases (ADH are a class of enzymes that catalyze the reversible oxidation of alcohols to corresponding aldehydes or ketones, by using either nicotinamide adenine dinucleotide (NAD or nicotinamide adenine dinucleotide phosphate (NADP, as coenzymes. In this study, a short-chain ADH gene was identified in Bombyx mori by 5'-RACE PCR. This is the first time the coding region of BmADH has been cloned, expressed, purified and then characterized. The cDNA fragment encoding the BmADH protein was amplified from a pool of silkworm cDNAs by PCR, and then cloned into E. coli expression vector pET-30a(+. The recombinant His-tagged BmADH protein was expressed in E. coli BL21 (DE3, and then purified by metal chelating affinity chromatography. The soluble recombinant BmADH, produced at low-growth temperature, was instrumental in catalyzing the ethanol-dependent reduction of NAD+, thereby indicating ethanol as one of the substrates of BmADH.

  9. Glucuronoarabinoxylans from sorghum grain

    NARCIS (Netherlands)

    Verbruggen, M.A.

    1996-01-01


    Water-unextractable cell wall materials (WUS) were prepared from raw, polished, and malted sorghum ( Sorghum vulgare cv. Fara Fara). Except for the amounts, hardly any difference could be observed between the WUS of these three raw materials. This means that cell wall

  10. Comparison of brown midrib-6 and -18 forage sorghum with conventional sorghum and corn silage in diets of lactating dairy cows.

    Science.gov (United States)

    Oliver, A L; Grant, R J; Pedersen, J F; O'Rear, J

    2004-03-01

    Total mixed rations containing conventional forage sorghum, brown midrib (bmr)-6 forage sorghum, bmr-18 forage sorghum, or corn silage were fed to Holstein dairy cows to determine the effect on lactation, ruminal fermentation, and total tract nutrient digestion. Sixteen multiparous cows (4 ruminally fistulated; 124 d in milk) were assigned to 1 of 4 diets in a replicated Latin square design with 4-wk periods (21-d adaptation and 7 d of collection). Diets consisted of 40% test silage, 10% alfalfa silage, and 50% concentrate mix (dry basis). Acid detergent lignin concentration was reduced by 21 and 13%, respectively, for the bmr-6 and bmr-18 sorghum silages when compared with the conventional sorghum. Dry matter intake was not affected by diet. Production of 4% fat-corrected milk was greatest for cows fed bmr-6 (33.7 kg/d) and corn silage (33.3 kg/d), was least for cows fed the conventional sorghum (29.1 kg/d), and was intermediate for cows fed the bmr-18 sorghum (31.2 kg/d), which did not differ from any other diet. Total tract neutral detergent fiber (NDF) digestibility was greatest for the bmr-6 sorghum (54.4%) and corn silage (54.1%) diets and was lower for the conventional (40.8%) and bmr-18 sorghum (47.9%) diets. In situ extent of NDF digestion was greatest for the bmr-6 sorghum (76.4%) and corn silage (79.0%) diets, least for the conventional sorghum diet (70.4%), and intermediate for the bmr-18 sorghum silage diet (73.1%), which was not different from the other diets. Results of this study indicate that the bmr-6 sorghum hybrid outperformed the conventional sorghum hybrid; the bmr-18 sorghum was intermediate between conventional and bmr-6 in most cases. Additionally, the bmr-6 hybrid resulted in lactational performance equivalent to the corn hybrid used in this study. There are important compositional differences among bmr forage sorghum hybrids that need to be characterized to predict animal response accurately.

  11. Cloning of transgenic tobacco BY-2 cells; an efficient method to analyse and reduce high natural heterogeneity of transgene expression.

    Science.gov (United States)

    Nocarova, Eva; Fischer, Lukas

    2009-04-22

    Phenotypic characterization of transgenic cell lines, frequently used in plant biology studies, is complicated because transgene expression in individual cells is often heterogeneous and unstable. To identify the sources and to reduce this heterogeneity, we transformed tobacco (Nicotiana tabacum L.) BY-2 cells with a gene encoding green fluorescent protein (GFP) using Agrobacterium tumefaciens, and then introduced a simple cloning procedure to generate cell lines derived from the individual transformed cells. Expression of the transgene was monitored by analysing GFP fluorescence in the cloned lines and also in lines obtained directly after transformation. The majority ( approximately 90%) of suspension culture lines derived from calli that were obtained directly from transformation consisted of cells with various levels of GFP fluorescence. In contrast, nearly 50% of lines generated by cloning cells from the primary heterogeneous suspensions consisted of cells with homogenous GFP fluorescence. The rest of the lines exhibited "permanent heterogeneity" that could not be resolved by cloning. The extent of fluorescence heterogeneity often varied, even among genetically identical clones derived from the primary transformed lines. In contrast, the offspring of subsequent cloning of the cloned lines was uniform, showing GFP fluorescence intensity and heterogeneity that corresponded to the original clone. The results demonstrate that, besides genetic heterogeneity detected in some lines, the primary lines often contained a mixture of epigenetically different cells that could be separated by cloning. This indicates that a single integration event frequently results in various heritable expression patterns, which are probably accidental and become stabilized in the offspring of the primary transformed cells early after the integration event. Because heterogeneity in transgene expression has proven to be a serious problem, it is highly advisable to use transgenes tagged with

  12. The Sorghum Gene for Leaf Color Changes upon Wounding (P Encodes a Flavanone 4-Reductase in the 3-Deoxyanthocyanidin Biosynthesis Pathway

    Directory of Open Access Journals (Sweden)

    Hiroyuki Kawahigashi

    2016-05-01

    Full Text Available Upon wounding or pathogen invasion, leaves of sorghum [Sorghum bicolor (L. Moench] plants with the P gene turn purple, whereas leaves with the recessive allele turn brown or tan. This purple phenotype is determined by the production of two 3-deoxyanthocyanidins, apigeninidin and luteolinidin, which are not produced by the tan-phenotype plants. Using map-based cloning in progeny from a cross between purple Nakei-MS3B (PP and tan Greenleaf (pp cultivars, we isolated this gene, which was located in a 27-kb genomic region around the 58.1 Mb position on chromosome 6. Four candidate genes identified in this region were similar to the maize leucoanthocyanidin reductase gene. None of them was expressed before wounding, and only the Sb06g029550 gene was induced in both cultivars after wounding. The Sb06g029550 protein was detected in Nakei-MS3B, but only slightly in Greenleaf, in which it may be unstable because of a Cys252Tyr substitution. A recombinant Sb06g029550 protein had a specific flavanone 4-reductase activity, and converted flavanones (naringenin or eriodictyol to flavan-4-ols (apiforol or luteoforol in vitro. Our data indicate that the Sb06g029550 gene is involved in the 3-deoxyanthocyanidin synthesis pathway.

  13. Nutritional, functional and rheological properties of processed sorghum and ragi grains

    Directory of Open Access Journals (Sweden)

    Himadri Mahajan

    2015-12-01

    Full Text Available This study was undertaken to determine the effect of different treatments such as roasting, puffing and germination of white sorghum, red sorghum and ragi grains on physicochemical, antioxidant, protein, amylose, bulk density, colour index and rheological properties of respective flour. In case of ragi, after roasting treatment, total phenolic content (TPC content for flour was increased from 0.331 ± 0.001 to 0.373 ± 0.004 mg of gallic acid equivalents per gram of dry sample. However, total flavonoids content was also increased slightly after different processing treatments. The rheological properties of respective flour were studied using Chopin Mixolab, where wheat flour dough profile acts as a reference to study the effect of different treatments. Dough elasticity which is expressed by the values of amplitude (Nm was found to be low in case of untreated flour as compared to wheat flour dough. Elasticity values of untreated flour such as white sorghum, red sorghum and ragi were 0.02, 0.00 and 0.06 Nm, respectively. Whereas, after processing treatments, values of elasticity for roasted flour dough of white sorghum, puffed flour dough of red sorghum and roasted flour dough of ragi increased to 0.36, 0.11 and 0.15 Nm, respectively, as compared to wheat flour dough of 0.10 Nm. The results found that roasted ragi flour had higher rate of starch gelatinization, lower starch retrogradation, high antioxidant and amylose contents which were found to be prospective ingredients in whole wheat flour in various baked and fermented food applications.

  14. Physical and Mechanical Properties of Sorghum Grains (Sorghum Vulgare

    Directory of Open Access Journals (Sweden)

    2016-11-01

    Full Text Available The physical and mechanical properties of sorghum grains (sorghum vulgare were studied at varying moisture contents of 13%, 20% and 30% (w.b. The four varieties of sorghum grains studied include; Dura, Guinea, Faterita and Kafir. Results indicate that the size ranges were 3.94mm - 4.83mm for Dura variety; 3.75mm - 4.54mm for Guinea variety; 3.21mm - 4.42mm for Kafir variety and 2.70mm - 4.14mm for Faterita variety. Irregularities in the shapes of the grains were observed but all approximated to a sphere. In the mechanical properties, at major diameter, Dura variety had highest rupture force of 1.16kN at 13% moisture content (w.b while the Guinea variety had the lowest rupture force of 0.955kN. In minor diameter, the Dura variety also recorded highest rupture force of 1.12kN at 13% moisture content (w.b while the Kafir variety had the lowest value of 0.952kN. Also at 20% moisture content, the Dura variety had highest rupture force of 1.025kN while the Guinea variety had the lowest rupture force of 0.965kN. The same trend applies in the varieties at 30% moisture content. This is because, increase in moisture content results to decrease in rupture force. And this implies that force beyond these points at these moisture contents may cause damage to the sorghum varieties.

  15. Field damage of sorghum (Sorghum bicolor) with reduced lignin levels by naturally occurring insect pests and pathogens

    Science.gov (United States)

    Mutant lines of sorghum with low levels of lignin are potentially useful for bioenergy production, but may have problems with insects or disease. Field grown normal and low lignin bmr6 and bmr12 sorghum (Sorghum bicolor) were examined for insect and disease damage in the field, and insect damage in ...

  16. (Arachis hypogaea) and Sorghum (Sorghum bicolor)

    African Journals Online (AJOL)

    ADOWIE PERE

    as enzyme activities of Arachis hypogaea and Sorghum bicolor in crude oil contaminated soil. Crude oil ... Treatments without crude oil were ... replicates were made for each treatment. .... dead sections of leaf margins, burning and stunted or.

  17. Fermentation characteristics of different purpose sorghum silage

    Directory of Open Access Journals (Sweden)

    Arthur Behling Neto

    2017-08-01

    Full Text Available Sorghum stands out among other plants recommended for ensiling due to its forage composition, its resistance to drought, and its planting range. New cultivars of grain and sweet sorghum that can be used for silage production are available, but there is little information regarding their ensiling characteristics. The aim of this study was to evaluate the fermentation characteristics at the ensiling of different purpose sorghum cultivars, at two crop periods. The trial was carried out at the Plant Production Department of the Federal Institute of Education, Science and Technology of Rondônia, Colorado do Oeste campus, Rondônia, Brazil, and chemical analyses were performed at the Laboratory of Animal Nutrition, at the Federal University of Mato Grosso, Cuiabá campus, Mato Grosso, Brazil. The experimental design used was a randomized block, in split-plot design, with four replicates. The plot treatments consisted of six sorghum cultivars grown for different purposes (grain sorghum: BRS 308 and BRS 310; forage sorghum: BR 655 and BRS 610; sweet sorghum: BRS 506 and CMSXS 647. Split-plot treatments consisted of two cropping seasons (first crop and second crop. The grain sorghum cultivar BRS 310 was the only one that had suitable dry matter content for ensiling; however, it was also the only one that did not show ideal water soluble carbohydrate content for ensiling. Nevertheless, all treatments presented pH below than 4.2 and ammonia nitrogen lower than 12% of total N, which indicates that the fermentation inside the silo had proceeded well. For sweet sorghum cultivars, higher ethanol and butyric acid content were observed for the first crop than for the second crop. All evaluated sorghum cultivars can be used for silage production, but the use of sweet sorghum is recommended at the second crop.

  18. Back to Acid Soil Fields: The Citrate Transporter SbMATE Is a Major Asset for Sustainable Grain Yield for Sorghum Cultivated on Acid Soils

    Directory of Open Access Journals (Sweden)

    Geraldo Carvalho Jr

    2016-02-01

    Full Text Available Aluminum (Al toxicity damages plant roots and limits crop production on acid soils, which comprise up to 50% of the world’s arable lands. A major Al tolerance locus on chromosome 3, AltSB, controls aluminum tolerance in sorghum [Sorghum bicolor (L. Moench] via SbMATE, an Al-activated plasma membrane transporter that mediates Al exclusion from sensitive regions in the root apex. As is the case with other known Al tolerance genes, SbMATE was cloned based on studies conducted under controlled environmental conditions, in nutrient solution. Therefore, its impact on grain yield on acid soils remains undetermined. To determine the real world impact of SbMATE, multi-trait quantitative trait loci (QTL mapping in hydroponics, and, in the field, revealed a large-effect QTL colocalized with the Al tolerance locus AltSB, where SbMATE lies, conferring a 0.6 ton ha–1 grain yield increase on acid soils. A second QTL for Al tolerance in hydroponics, where the positive allele was also donated by the Al tolerant parent, SC283, was found on chromosome 9, indicating the presence of distinct Al tolerance genes in the sorghum genome, or genes acting in the SbMATE pathway leading to Al-activated citrate release. There was no yield penalty for AltSB, consistent with the highly localized Al regulated SbMATE expression in the root tip, and Al-dependent transport activity. A female effect of 0.5 ton ha–1 independently demonstrated the effectiveness of AltSB in hybrids. Al tolerance conferred by AltSB is thus an indispensable asset for sorghum production and food security on acid soils, many of which are located in developing countries.

  19. Cloning and expression in Escherichia coli of cellulases genes from Clostridium IBUN 22A

    Directory of Open Access Journals (Sweden)

    Lucy Carolina Vargas Pabón

    2002-01-01

    Full Text Available Genomic library of the native strain Clostridium IBUN 22A was constructed, using plasmid pBluescriptlI® KS+/ - as cloning vector and its expression in Escherichia coli was evaluated. Eight recombination clones with enzymatic activity were detected by enzymatic screening and using the red-Congo test with three substrates: cellobiose, carboxymethyl cellulose (CMC and cellulose powder (native. Restriction analysis of three recombination plasmids, representative of each enzymatic activity showed the inserted size (1600, 13000 and 11000bp approximately for pBS68, pBS25 and pBS57 respectively. More studies of protein expression and enzymatic characterization will allow theses enzymes and other typical parameters to be defined. In the same way the fragment sequence cloned will lead to a more detailed analysis and definition of the biotechnological potential of this strain regarding solvent production using cellulosic substrates for fermentation.

  20. Nutritional value of sorghum silage of different purposes

    Directory of Open Access Journals (Sweden)

    Arthur Behling Neto

    Full Text Available ABSTRACT Sorghum is a crop that stands out as an alternative to corn due to lower soil fertility demand and increased tolerance to drought. Lack of information about the qualitative behaviour of sorghum hinders the recommendation of different purpose sorghum cultivars. The goal was to evaluate the chemical composition and in vitro digestibility of different purpose sorghum cultivar silages, at two cropping seasons. The trial was conducted at the Plant Production Department, Federal Institute of Education, Science and Technology of Rondônia, Colorado do Oeste campus, and chemical analyses and in vitro incubation were performed at the Laboratory of Animal Nutrition, Federal University of Mato Grosso, Cuiabá campus. The experimental design was a randomized block with a split-plot arrangement and four replications. Plot treatments consisted of six different purpose sorghum cultivars (BRS 308 and BRS 310, grain sorghum; BR 655 and BRS 610, forage sorghum; and BRS 506 and CMSXS 647, sweet sorghum. Split-plot treatments consisted of two cropping periods (first crop and second crop. Forage sorghum cultivar BRS 655 demonstrated higher non-fiber carbohydrate content and lower potentially digestible fibre content than the other cultivars did. Sweet sorghum cultivars had higher levels of water soluble carbohydrates and non-protein nitrogen based on protein, lower indigestible neutral detergent fibre content at second crop, and higher in vitro dry matter digestibility than the other cultivars. The silages of sweet sorghum cultivars BRS 506 and CMSXS 647, and forage sorghum cultivar BRS 655 presented higher nutritional values.

  1. A plasmid toolkit for cloning chimeric cDNAs encoding customized fusion proteins into any Gateway destination expression vector

    Science.gov (United States)

    2013-01-01

    Background Valuable clone collections encoding the complete ORFeomes for some model organisms have been constructed following the completion of their genome sequencing projects. These libraries are based on Gateway cloning technology, which facilitates the study of protein function by simplifying the subcloning of open reading frames (ORF) into any suitable destination vector. The expression of proteins of interest as fusions with functional modules is a frequent approach in their initial functional characterization. A limited number of Gateway destination expression vectors allow the construction of fusion proteins from ORFeome-derived sequences, but they are restricted to the possibilities offered by their inbuilt functional modules and their pre-defined model organism-specificity. Thus, the availability of cloning systems that overcome these limitations would be highly advantageous. Results We present a versatile cloning toolkit for constructing fully-customizable three-part fusion proteins based on the MultiSite Gateway cloning system. The fusion protein components are encoded in the three plasmids integral to the kit. These can recombine with any purposely-engineered destination vector that uses a heterologous promoter external to the Gateway cassette, leading to the in-frame cloning of an ORF of interest flanked by two functional modules. In contrast to previous systems, a third part becomes available for peptide-encoding as it no longer needs to contain a promoter, resulting in an increased number of possible fusion combinations. We have constructed the kit’s component plasmids and demonstrate its functionality by providing proof-of-principle data on the expression of prototype fluorescent fusions in transiently-transfected cells. Conclusions We have developed a toolkit for creating fusion proteins with customized N- and C-term modules from Gateway entry clones encoding ORFs of interest. Importantly, our method allows entry clones obtained from ORFeome

  2. Identification and profiling of salinity stress-responsive proteins in Sorghum bicolor seedlings

    DEFF Research Database (Denmark)

    Ngara, Rudo; Ndimba, Roya; Borch-Jensen, Jonas

    2012-01-01

    Sorghum bicolor, a drought tolerant cereal crop, is not only an important food source in the semi arid/arid regions but also a potential model for studying and gaining a better understanding of the molecular mechanisms of drought and salt stress tolerance in cereals. In this study, seeds of a sweet...... sorghum variety, MN1618, were planted and grown on solid MS growth medium with or without 100mM NaCl. Heat shock protein expression immunoblotting assays demonstrated that this salt treatment induced stress within natural physiological parameters for our experimental material. 2D PAGE in combination...... with MS/MS proteomics techniques were used to separate, visualise and identify salinity stress responsive proteins in young sorghum leaves. Out of 281 Coomassie stainable spots, 118 showed statistically significant responses (p...

  3. Development of Perennial Grain Sorghum

    Directory of Open Access Journals (Sweden)

    Stan Cox

    2018-01-01

    Full Text Available Perennial germplasm derived from crosses between Sorghum bicolor and either S. halepense or S. propinquum is being developed with the goal of preventing and reversing soil degradation in the world’s grain sorghum-growing regions. Perennial grain sorghum plants produce subterranean stems known as rhizomes that sprout to form the next season’s crop. In Kansas, breeding perennial sorghum involves crossing S. bicolor cultivars or breeding lines to S. halepense or perennial S. bicolorn × S. halepense breeding lines, selecting perennial plants from F2 or subsequent populations, crossing those plants with S. bicolor, and repeating the cycle. A retrospective field trial in Kansas showed that selection and backcrossing during 2002–2009 had improved grain yields and seed weights of breeding lines. Second-season grain yields of sorghum lines regrowing from rhizomes were similar to yields in the first season. Further selection cycles have been completed since 2009. Many rhizomatous lines that cannot survive winters in Kansas are perennial at subtropical or tropical locations in North America and Africa. Grain yield in Kansas was not correlated with rhizomatousness in either Kansas or Uganda. Genomic regions affecting rhizome growth and development have been mapped, providing new breeding tools. The S. halepense gene pool may harbor many alleles useful for improving sorghum for a broad range of traits in addition to perenniality.

  4. Global analysis of epigenetic regulation of gene expression in response to drought stress in Sorghum.

    Energy Technology Data Exchange (ETDEWEB)

    Reddy, Anireddy [Colorado State Univ., Fort Collins, CO (United States); Ben-Hur, Asa [Colorado State Univ., Fort Collins, CO (United States)

    2017-11-22

    Abiotic stresses including drought are major limiting factors of crop yields and cause significant crop losses. Acquisition of stress tolerance to abiotic stresses requires coordinated regulation of a multitude of biochemical and physiological changes, and most of these changes depend on alterations in gene expression. The goal of this work is to perform global analysis of differential regulation of gene expression and alternative splicing, and their relationship with chromatin landscape in drought sensitive and tolerant cultivars. our Iso-Seq study revealed transcriptome-wide full-length isoforms at an unprecedented scale with over 11000 novel splice isoforms. Additionally, we uncovered alternative polyadenylation sites of ~11000 expressed genes and many novel genes. Overall, Iso-Seq results greatly enhanced sorghum gene annotations that are not only useful in analyzing all our RNA-seq, ChIP-seq and ATAC-seq data but also serve as a great resource to the plant biology community. Our studies identified differentially expressed genes and splicing events that are correlated with the drought-resistant phenotype. An association between alternative splicing and chromatin accessibility was also revealed. Several computational tools developed here (TAPIS and iDiffIR) have been made freely available to the research community in analyzing alternative splicing and differential alternative splicing.

  5. In vitro binding of Sorghum bicolor transcription factors ABI4 and ABI5 to a conserved region of a GA 2-OXIDASE promoter: possible role of this interaction in the expression of seed dormancy.

    Science.gov (United States)

    Cantoro, Renata; Crocco, Carlos Daniel; Benech-Arnold, Roberto Luis; Rodríguez, María Verónica

    2013-12-01

    The precise adjustment of the timing of dormancy release according to final grain usage is still a challenge for many cereal crops. Grain sorghum [Sorghum bicolor (L.) Moench] shows wide intraspecific variability in dormancy level and susceptibility to pre-harvest sprouting (PHS). Both embryo sensitivity to abscisic acid (ABA) and gibberellin (GA) metabolism play an important role in the expression of dormancy of the developing sorghum grain. In previous works, it was shown that, simultaneously with a greater embryo sensitivity to ABA and higher expression of SbABA-INSENSITIVE 4 (SbABI4) and SbABA-INSENSITIVE 5 (SbABI5), dormant grains accumulate less active GA4 due to a more active GA catabolism. In this work, it is demonstrated that the ABA signalling components SbABI4 and SbABI5 interact in vitro with a fragment of the SbGA 2-OXIDASE 3 (SbGA2ox3) promoter containing an ABA-responsive complex (ABRC). Both transcription factors were able to bind the promoter, although not simultaneously, suggesting that they might compete for the same cis-acting regulatory sequences. A biological role for these interactions in the expression of dormancy of sorghum grains is proposed: either SbABI4 and/or SbABI5 activate transcription of the SbGA2ox3 gene in vivo and promote SbGA2ox3 protein accumulation; this would result in active degradation of GA4, thus preventing germination of dormant grains. A comparative analysis of the 5'-regulatory region of GA2oxs from both monocots and dicots is also presented; conservation of the ABRC in closely related GA2oxs from Brachypodium distachyon and rice suggest that these species might share the same regulatory mechanism as proposed for grain sorghum.

  6. Cloning and expression analysis of two dehydrodolichyl diphosphate synthase genes from Tripterygium wilfordii

    Directory of Open Access Journals (Sweden)

    Lin-Hui Gao

    2018-01-01

    Full Text Available Objective: To clone and investigate two dehydrodolichyl diphosphate synthase genes of Tripterygium wilfordii by bioinformatics and tissue expression analysis. Materials and Methods: According to the T. wifordii transcriptome database, specific primers were designed to clone the TwDHDDS1 and TwDHDDS2 genes via PCR. Based on the cloned sequences, protein structure prediction, multiple sequence alignment and phylogenetic tree construction were performed. The expression levels of the genes in different tissues of T. wilfordii were measured by real-time quantitative PCR. Results: The TwDHDDS1 gene encompassed a 873 bp open reading frame (ORF and encoded a protein of 290 amino acids. The calculated molecular weight of the translated protein was about 33.46 kDa, and the theoretical isoelectric point (pI was 8.67. The TwDHDDS2 encompassed a 768 bp ORF, encoding a protein of 255 amino acids with a calculated molecular weight of about 21.19 kDa, and a theoretical isoelectric point (pI of 7.72. Plant tissue expression analysis indicated that TwDHDDS1 and TwDHDDS2 both have relatively ubiquitous expression in all sampled organ tissues, but showed the highest transcription levels in the stems. Conclusions: The results of this study provide a basis for further functional studies of TwDHDDS1 and TwDHDDS2. Most importantly, these genes are promising genetic targets for the regulation of the biosynthetic pathways of important bioactive terpenoids such as triptolide.

  7. Cloning and prokaryotic expression of the porcine lipasin gene.

    Science.gov (United States)

    Li, M M; Geng, J; Guo, Y J; Jiao, X Q; Lu, W F; Zhu, H S; Wang, Y Y; Yang, G Y

    2015-11-23

    Lipasin has recently been demonstrated to be involved in lipid metabolism. In this study, two specific primers were used to amplify the lipasin open reading frame from porcine liver tissue. The polymerase chain reaction product was cloned to a pGEM®-T Easy Vector, digested by SalI and NotI, and sequenced. The lipasin fragment was then cloned to a pET21(b) vector and digested by the same restriction enzyme. The recombinant plasmid was transferred to Escherichia coli (BL21), and the lipasin protein was induced with isopropyl-β-D-thiogalactopyranoside. The protein obtained was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis and western blotting. A pET-lipasin prokaryotic recombinant expression vector was successfully constructed, and a 25.2-kDa protein was obtained. This study provides a basis for further research on the biological function of porcine lipasin.

  8. Global Expression in Sorghum Brown Midrib Mutants to Improve Biomass for Biofuels

    Science.gov (United States)

    Brown midrib (bmr) mutants are being investigated for their ability to increase the conversion efficiency of sorghum biomass for lignocellulosic bioenergy. Brown midrib 6 and 12 (bmr6 and 12) are impaired the last two steps of monolignol biosynthesis resulting in reduced lignin content and altered ...

  9. Cloning and expression of Tenebrio molitor antifreeze protein in Escherichia coli.

    Science.gov (United States)

    Yue, Chang-Wu; Zhang, Yi-Zheng

    2009-03-01

    A novel antifreeze protein cDNA was cloned by RT-PCR from the larva of the yellow mealworm Tenebrio molitor. The coding fragment of 339 bp encodes a protein of 112 amino acid residues and was fused to the expression vectors pET32a and pTWIN1. The resulted expression plasmids were transformed into Escherischia coli strains BL21 (DE3), ER2566, and Origami B (DE3), respectively. Several strategies were used for expression of the highly disulfide-bonded beta-helix-contained protein with the activity of antifreeze in different expression systems. A protocol for production of refolded and active T. molitor antifreeze protein in bacteria was obtained.

  10. A versatile system for USER cloning-based assembly of expression vectors for mammalian cell engineering.

    Directory of Open Access Journals (Sweden)

    Anne Mathilde Lund

    Full Text Available A new versatile mammalian vector system for protein production, cell biology analyses, and cell factory engineering was developed. The vector system applies the ligation-free uracil-excision based technique--USER cloning--to rapidly construct mammalian expression vectors of multiple DNA fragments and with maximum flexibility, both for choice of vector backbone and cargo. The vector system includes a set of basic vectors and a toolbox containing a multitude of DNA building blocks including promoters, terminators, selectable marker- and reporter genes, and sequences encoding an internal ribosome entry site, cellular localization signals and epitope- and purification tags. Building blocks in the toolbox can be easily combined as they contain defined and tested Flexible Assembly Sequence Tags, FASTs. USER cloning with FASTs allows rapid swaps of gene, promoter or selection marker in existing plasmids and simple construction of vectors encoding proteins, which are fused to fluorescence-, purification-, localization-, or epitope tags. The mammalian expression vector assembly platform currently allows for the assembly of up to seven fragments in a single cloning step with correct directionality and with a cloning efficiency above 90%. The functionality of basic vectors for FAST assembly was tested and validated by transient expression of fluorescent model proteins in CHO, U-2-OS and HEK293 cell lines. In this test, we included many of the most common vector elements for heterologous gene expression in mammalian cells, in addition the system is fully extendable by other users. The vector system is designed to facilitate high-throughput genome-scale studies of mammalian cells, such as the newly sequenced CHO cell lines, through the ability to rapidly generate high-fidelity assembly of customizable gene expression vectors.

  11. Cloning and expression of Pectobacterium carotovorum endo-polygalacturonase gene in Pichia pastoris for production of oligogalacturonates

    Science.gov (United States)

    A bacterial endo-polygalacturonase (endo-PGase) gene from the plant pathogen Pectobacterium carotovorum was cloned into pGAPZaA vector and constitutively expressed in Pichia pastoris. The recombinant endo-PGase secreted by the Pichia clone showed a 1.7 fold increase when the culture medium included ...

  12. Sorghum yield and associated satellite-derived meteorological ...

    African Journals Online (AJOL)

    Sorghum yield and associated satellite-derived meteorological parameters in semi-arid Botswana. ... African Crop Science Journal ... Sorghum (Sorghum bicolor) yield for five seasons (2005/6 to 2009/10) from the Botswana Department of Crop ... Key Words: Coefficient of determination, NDVI, Pearson correlation ...

  13. Cloning of a Gene Whose Expression is Increased in Scrapie and in Senile Plaques in Human Brain

    Science.gov (United States)

    Wietgrefe, S.; Zupancic, M.; Haase, A.; Chesebro, B.; Race, R.; Frey, W.; Rustan, T.; Friedman, R. L.

    1985-12-01

    A complementary DNA library was constructed from messenger RNA's extracted from the brains of mice infected with the scrapie agent. The library was differentially screened with the objectives of finding clones that might be used as markers of infection and finding clones of genes whose increased expression might be correlated with the pathological changes common to scrapie and Alzheimer's disease. A gene was identified whose expression is increased in scrapie. The complementary DNA corresponding to this gene hybridized preferentially and focally to cells in the brains of scrapie-infected animals. The cloned DNA also hybridized to the neuritic plaques found with increased frequency in brains of patients with Alzheimer's disease.

  14. Cloning and heterologous expression of a novel insecticidal gene (tccC1) from Xenorhabdus nematophilus strain

    International Nuclear Information System (INIS)

    Joo Lee, Pom; Ahn, Ji-Young; Kim, Yang-Hoon; Wook Kim, Seung; Kim, Ji-Yeon; Park, Jae-Sung; Lee, Jeewon

    2004-01-01

    We have identified and cloned a novel toxin gene (tccC1/xptB1) from Xenorhabdus nematophilus strain isolated from Korea-specific entomophagous nematode Steinernema glaseri MK. The DNA sequence of cloned toxin gene (3048 bp) has an open reading frame encoding 1016 amino acids with a predicted molecular mass of 111058 Da. The toxin sequence shares 50-96% identical amino acid residues with the previously reported tccC1 cloned from X. nematophilus (AJ308438), Photorhabdus luminescens W14 (AF346499) P. luminescens TTO1 (BX571873), and Yersinia pestis CO92 (NC 0 03143). The toxin gene was successfully expressed in Escherichia coli, and the recombinant toxin protein caused a rapid cessation in mortality of Galleria mellonella larvae (80% death of larvae within 2 days). Conclusively, the heterologous expression of the novel gene tccC1 cloned into E. coli plasmid vector produced recombinant toxin with high insecticidal activity

  15. Cloning of a yeast alpha-amylase promoter and its regulated heterologous expression

    Science.gov (United States)

    Gao, Johnway [Richland, WA; Skeen, Rodney S [Pendleton, OR; Hooker, Brian S [Kennewick, WA; Anderson, Daniel B [Pasco, WA

    2003-04-01

    The present invention provides the promoter clone discovery of an alpha-amylase gene of a starch utilizing yeast strain Schwanniomyces castellii. The isolated alpha-amylase promoter is an inducible promoter, which can regulate strong gene expression in starch culture medium.

  16. Cross-species multiple environmental stress responses: An integrated approach to identify candidate genes for multiple stress tolerance in sorghum (Sorghum bicolor (L. Moench and related model species.

    Directory of Open Access Journals (Sweden)

    Adugna Abdi Woldesemayat

    Full Text Available Crop response to the changing climate and unpredictable effects of global warming with adverse conditions such as drought stress has brought concerns about food security to the fore; crop yield loss is a major cause of concern in this regard. Identification of genes with multiple responses across environmental stresses is the genetic foundation that leads to crop adaptation to environmental perturbations.In this paper, we introduce an integrated approach to assess candidate genes for multiple stress responses across-species. The approach combines ontology based semantic data integration with expression profiling, comparative genomics, phylogenomics, functional gene enrichment and gene enrichment network analysis to identify genes associated with plant stress phenotypes. Five different ontologies, viz., Gene Ontology (GO, Trait Ontology (TO, Plant Ontology (PO, Growth Ontology (GRO and Environment Ontology (EO were used to semantically integrate drought related information.Target genes linked to Quantitative Trait Loci (QTLs controlling yield and stress tolerance in sorghum (Sorghum bicolor (L. Moench and closely related species were identified. Based on the enriched GO terms of the biological processes, 1116 sorghum genes with potential responses to 5 different stresses, such as drought (18%, salt (32%, cold (20%, heat (8% and oxidative stress (25% were identified to be over-expressed. Out of 169 sorghum drought responsive QTLs associated genes that were identified based on expression datasets, 56% were shown to have multiple stress responses. On the other hand, out of 168 additional genes that have been evaluated for orthologous pairs, 90% were conserved across species for drought tolerance. Over 50% of identified maize and rice genes were responsive to drought and salt stresses and were co-located within multifunctional QTLs. Among the total identified multi-stress responsive genes, 272 targets were shown to be co-localized within QTLs

  17. Cloning and Expression of Listeria monocytogenes Listeriolysin O in Lactobacillus plantarum

    Directory of Open Access Journals (Sweden)

    Masoumeh Hayati

    2017-11-01

    Full Text Available Background: The protein listeriolysin O (LLO encoded by hly gene, is one of the most important virulence factors of Listeria monocytogenes. This highly potent immunogenic cholesterol binding toxin has hemolytic activity, responsible for phagosomal membrane disruption and bacterial escape to the cytoplasm and facilitating the stimulation of CD8+ T cells and Th1 response. Recently pathobiotechnological vaccination using probiotic bacteria have been proposed. One of these strategies is expression of LLO in non-pathogenic bacteria such as lactic acid bacteria as delivery strains. Objectives: Our aim in this study was cloning of hly gene in a Lactobacillus species via pNZ8110, an inducible expression vector which is specific for Lactococcus species. Materials and Methods: hly gene was amplified by PCR and cloned into pNZ8110 by restriction enzymes cutting and ligation method. After transformation and propagation in E. coli MC1061 intermediate host, it was successfully electrotransformed into Lactobacillus plantarum. Results: Gel electrophoresis of colony PCR, extracted plasmids and restriction analysis along with sequencing confirmed the transformation. After induction using supernatant of nisin producer Lactococcus lactis NZ9700 strain, Expression of LLO was confirmed by SDS PAGE and western blot. Conclusion: Here, we have employed a nonpathogenic probiotic strain; Lactobacillus plantarum for the first time to express hly gene of Listeria monocytogenes in order to propose a new vaccine candidate.

  18. Genetic analysis of recombinant inbred lines for Sorghum bicolor × Sorghum propinquum.

    Science.gov (United States)

    Kong, Wenqian; Jin, Huizhe; Franks, Cleve D; Kim, Changsoo; Bandopadhyay, Rajib; Rana, Mukesh K; Auckland, Susan A; Goff, Valorie H; Rainville, Lisa K; Burow, Gloria B; Woodfin, Charles; Burke, John J; Paterson, Andrew H

    2013-01-01

    We describe a recombinant inbred line (RIL) population of 161 F5 genotypes for the widest euploid cross that can be made to cultivated sorghum (Sorghum bicolor) using conventional techniques, S. bicolor × Sorghum propinquum, that segregates for many traits related to plant architecture, growth and development, reproduction, and life history. The genetic map of the S. bicolor × S. propinquum RILs contains 141 loci on 10 linkage groups collectively spanning 773.1 cM. Although the genetic map has DNA marker density well-suited to quantitative trait loci mapping and samples most of the genome, our previous observations that sorghum pericentromeric heterochromatin is recalcitrant to recombination is highlighted by the finding that the vast majority of recombination in sorghum is concentrated in small regions of euchromatin that are distal to most chromosomes. The advancement of the RIL population in an environment to which the S. bicolor parent was well adapted (indeed bred for) but the S. propinquum parent was not largely eliminated an allele for short-day flowering that confounded many other traits, for example, permitting us to map new quantitative trait loci for flowering that previously eluded detection. Additional recombination that has accrued in the development of this RIL population also may have improved resolution of apices of heterozygote excess, accounting for their greater abundance in the F5 than the F2 generation. The S. bicolor × S. propinquum RIL population offers advantages over early-generation populations that will shed new light on genetic, environmental, and physiological/biochemical factors that regulate plant growth and development.

  19. Dhurrin content relates to sorghum (Sorghum bicolor (L) Moench) seedling growth in marginal soils.

    Science.gov (United States)

    Dhurrin content in leaves of mature sorghum plant is a quantitative measure of the level of pre-and postflowering drought tolerance (Burke et al., 2013). Postflowering drought tolerance in sorghum is linked to the staygreen (delayed senescence) trait (Howarth, 2000; Rosenow et al., 1977) which has b...

  20. Dhurrin content relates to sorghum [sorghum bicolor (L.) Moench] seedling growth in marginal soils

    Science.gov (United States)

    Dhurrin content in leaves of mature sorghum plant is a quantitative measure of the level of pre-and postflowering drought tolerance (Burke et al., 2013). Postflowering drought tolerance in sorghum is linked to the staygreen (delayed senescence) trait (Howarth, 2000; Rosenow et al., 1977) which has ...

  1. Cloning-free regulated monitoring of reporter and gene expression

    Directory of Open Access Journals (Sweden)

    Demirkaya Omer

    2009-03-01

    Full Text Available Abstract Background The majority of the promoters, their regulatory elements, and their variations in the human genome remain unknown. Reporter gene technology for transcriptional activity is a widely used tool for the study of promoter structure, gene regulation, and signaling pathways. Construction of transcriptional reporter vectors, including use of cis-acting sequences, requires cloning and time-demanding manipulations, particularly with introduced mutations. Results In this report, we describe a cloning-free strategy to generate transcriptionally-controllable linear reporter constructs. This approach was applied in common transcriptional models of inflammatory response and the interferon system. In addition, it was used to delineate minimal transcriptional activity of selected ribosomal protein promoters. The approach was tested for conversion of genes into TetO-inducible/repressible expression cassettes. Conclusion The simple introduction and tuning of any transcriptional control in the linear DNA product renders promoter activation and regulated gene studies simple and versatile.

  2. Cloning and Expressing Recombinant Protective Antigen Domains of B. anthracis

    Science.gov (United States)

    2011-09-01

    future predictive modeling toolkits. 1 1. Introduction The use of Bacillus anthracis as a bio - weapon in the United States in 2001 affirmed the need...for improved sensing and detection of biological weapons of mass destruction (WMD). Protective Antigen (PA) protein of Bacillus anthracis is the...Cloning and Expressing Recombinant Protective Antigen Domains of B. anthracis by Deborah A. Sarkes, Joshua M. Kogot, Irene Val-Addo

  3. Plant resistance in sorghums to the sugarcane aphid Melanaphis sacchari (Hemiptera: Aphididae)

    Science.gov (United States)

    We evaluated ten sorghum lines that were near or in commercial release with the intent of identifying phenotypic expression of host-plant resistance to the sugarcane aphid. Two of the ten entries OL2042 and SP7715 expressed a high degree of resistance to the sugarcane aphid with damage ratings <3.0...

  4. Cloning and Expression of Yak Active Chymosin in

    Directory of Open Access Journals (Sweden)

    Fan Luo

    2016-09-01

    Full Text Available Rennet, a complex of enzymes found in the stomachs of ruminants, is an important component for cheese production. In our study, we described that yak chymosin gene recombinant Pichia pastoris strain could serve as a novel source for rennet production. Yaks total RNA was extracted from the abomasum of an unweaned yak. The yak preprochymosin, prochymosin, and chymosin genes from total RNA were isolated using gene specific primers based on cattle chymosin gene sequence respectively and analyzed their expression pattern byreal time-polymerase chain reaction. The result showed that the chymosin gene expression level of the sucking yaks was 11.45 times higher than one of adult yaks and yak chymosin belongs to Bovidae family in phylogenetic analysis. To express each, the preprochymosin, prochymosin, and chymosin genes were ligated into the expression vector pPICZαA, respectively, and were expressed in Pichia pastoris X33. The results showed that all the recombinant clones of P. pastoris containing the preprochymosin, prochymosin or chymosin genes could produce the active form of recombinant chymosin into the culture supernatant. Heterologous expressed prochymosin (14.55 Soxhlet unit/mL had the highest enzyme activity of the three expressed chymosin enzymes. Therefore, we suggest that the yak chymosin gene recombinant Pichia pastoris strain could provide an alternative source of rennet production.

  5. Cloning, Codon Optimization, and Expression of Yersinia intermedia Phytase Gene in E. coli.

    Science.gov (United States)

    Mirzaei, Maryam; Saffar, Behnaz; Shareghi, Behzad

    2016-06-01

    Phytate is an anti-nutritional factor in plants, which catches the most phosphorus contents and some vital minerals. Therefore, Phytase is added mainly as an additive to the monogastric animals' foods to hydrolyze phytate and increase absorption of phosphorus. Y. intermedia phytase is a new phytase with special characteristics such as high specific activity, pH stability, and thermostability. Our aim was to clone, express, and characterizea codon optimized Y. intermedia phytase gene in E. coli . The Y. intermedia phytase gene was optimized according to the codon usage in E. coli . The sequence was synthesized and sub-cloned in pET-22b (+) vector and transformed into E. coli Bl21 (DE3). The protein was expressed in the presence of IPTG at a final concentration of 1 mM at 30°C. The purification of recombinant protein was performed by Ni 2+ affinity chromatography. Phytase activity and stability were determined in various pH and temperatures. The codon optimized Y. intermedia phytase gene was sub-cloned successfully.The expression was confirmed by SDS-PAGE and Western blot analysis. The recombinant enzyme (approximately 45 kDa) was purified. Specific activity of enzyme was 3849 (U.mg -1 ) with optimal pH 5 and optimal temperature of 55°C. Thermostability (80°C for 15 min) and pH stability (3-6) of the enzyme were 56 and more than 80%, respectively. The results of the expression and enzyme characterization revealed that the optimized Y. intermedia phytase gene has a good potential to be produced commercially andto be applied in animals' foodsindustry.

  6. Rate and Timing Effects of Growth Regulating Herbicides Applications on Grain Sorghum (Sorghum bicolor Growth and Yield

    Directory of Open Access Journals (Sweden)

    Thierry E. Besançon

    2016-01-01

    Full Text Available Dicamba and 2,4-D are among the most common and inexpensive herbicides used to control broadleaf weeds. However, different studies have pointed the risk of crop injury and grain sorghum yield reduction with postemergence applications of 2,4-D. No research data on grain sorghum response to 2,4-D or dicamba exists in the Southeastern United States. Consequently, a study was conducted to investigate crop growth and yield response to 2,4-D (100, 220, and 330 g acid equivalent ha−1 and dicamba (280 g acid equivalent ha−1 applied on 20 to 65 cm tall sorghum. Greater stunting resulted from 2,4-D applied at 330 g acid equivalent ha−1 or below 45 cm tall sorghum whereas lodging prevailed with 2,4-D at 330 g acid equivalent ha−1 and dicamba applied beyond 35 cm tall crop. Regardless of local environmental conditions, 2,4-D applied up to 35 cm tall did not negatively impact grain yield. There was a trend for yields to be somewhat lower when 2,4-D was applied on 45 or 55 cm tall sorghum whereas application on 65 cm tall sorghum systematically decreased yields. More caution should be taken with dicamba since yield reduction has been reported as early as applications made on 35 cm tall sorghum for a potentially dicamba sensitive cultivar.

  7. Isolation, molecular cloning and expression of cellobiohydrolase B (CbhB) from Aspergillus niger in Escherichia coli

    International Nuclear Information System (INIS)

    Woon, J. S. K.; Murad, A. M. A.; Abu Bakar, F. D.

    2015-01-01

    A cellobiohydrolase B (CbhB) from Aspergillus niger ATCC 10574 was cloned and expressed in E. coli. CbhB has an open reading frame of 1611 bp encoding a putative polypeptide of 536 amino acids. Analysis of the encoded polypeptide predicted a molecular mass of 56.2 kDa, a cellulose binding module (CBM) and a catalytic module. In order to obtain the mRNA of cbhB, total RNA was extracted from A. niger cells induced by 1% Avicel. First strand cDNA was synthesized from total RNA via reverse transcription. The full length cDNA of cbhB was amplified by PCR and cloned into the cloning vector, pGEM-T Easy. A comparison between genomic DNA and cDNA sequences of cbhB revealed that the gene is intronless. Upon the removal of the signal peptide, the cDNA of cbhB was cloned into the expression vector pET-32b. However, the recombinant CbhB was expressed in Escherichia coli Origami DE3 as an insoluble protein. A homology model of CbhB predicted the presence of nine disulfide bonds in the protein structure which may have contributed to the improper folding of the protein and thus, resulting in inclusion bodies in E. coli

  8. Isolation, molecular cloning and expression of cellobiohydrolase B (CbhB) from Aspergillus niger in Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    Woon, J. S. K., E-mail: jameswoon@siswa.ukm.edu.my; Murad, A. M. A., E-mail: munir@ukm.edu.my; Abu Bakar, F. D., E-mail: fabyff@ukm.edu.my [School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor (Malaysia)

    2015-09-25

    A cellobiohydrolase B (CbhB) from Aspergillus niger ATCC 10574 was cloned and expressed in E. coli. CbhB has an open reading frame of 1611 bp encoding a putative polypeptide of 536 amino acids. Analysis of the encoded polypeptide predicted a molecular mass of 56.2 kDa, a cellulose binding module (CBM) and a catalytic module. In order to obtain the mRNA of cbhB, total RNA was extracted from A. niger cells induced by 1% Avicel. First strand cDNA was synthesized from total RNA via reverse transcription. The full length cDNA of cbhB was amplified by PCR and cloned into the cloning vector, pGEM-T Easy. A comparison between genomic DNA and cDNA sequences of cbhB revealed that the gene is intronless. Upon the removal of the signal peptide, the cDNA of cbhB was cloned into the expression vector pET-32b. However, the recombinant CbhB was expressed in Escherichia coli Origami DE3 as an insoluble protein. A homology model of CbhB predicted the presence of nine disulfide bonds in the protein structure which may have contributed to the improper folding of the protein and thus, resulting in inclusion bodies in E. coli.

  9. Isolation, molecular cloning and expression of cellobiohydrolase B (CbhB) from Aspergillus niger in Escherichia coli

    Science.gov (United States)

    Woon, J. S. K.; Murad, A. M. A.; Abu Bakar, F. D.

    2015-09-01

    A cellobiohydrolase B (CbhB) from Aspergillus niger ATCC 10574 was cloned and expressed in E. coli. CbhB has an open reading frame of 1611 bp encoding a putative polypeptide of 536 amino acids. Analysis of the encoded polypeptide predicted a molecular mass of 56.2 kDa, a cellulose binding module (CBM) and a catalytic module. In order to obtain the mRNA of cbhB, total RNA was extracted from A. niger cells induced by 1% Avicel. First strand cDNA was synthesized from total RNA via reverse transcription. The full length cDNA of cbhB was amplified by PCR and cloned into the cloning vector, pGEM-T Easy. A comparison between genomic DNA and cDNA sequences of cbhB revealed that the gene is intronless. Upon the removal of the signal peptide, the cDNA of cbhB was cloned into the expression vector pET-32b. However, the recombinant CbhB was expressed in Escherichia coli Origami DE3 as an insoluble protein. A homology model of CbhB predicted the presence of nine disulfide bonds in the protein structure which may have contributed to the improper folding of the protein and thus, resulting in inclusion bodies in E. coli.

  10. Characterization of a pollen-specific cDNA clone from Nicotiana tabacum expressed during microgametogenesis and germination.

    Science.gov (United States)

    Weterings, K; Reijnen, W; van Aarssen, R; Kortstee, A; Spijkers, J; van Herpen, M; Schrauwen, J; Wullems, G

    1992-04-01

    This report describes the isolation and characterization of a cDNA clone representing a gene specifically expressed in pollen. A cDNA library was constructed against mRNA from mature pollen of Nicotiana tabacum. It was screened differentially against cDNA from mRNA of leaf and of pollen. One clone, NTPc303, was further characterized. On northern blot this clone hybridizes to a transcript 2100 nucleotides in length. NTPc303 is abundant in pollen. Expression of the corresponding gene is restricted to pollen, because no other generative or vegetative tissue contains transcripts hybridizing to NTPc303. Expression of NTP303 is evolutionarily conserved: homologous transcripts are present in pollen from various plant species. The first NTP303 transcripts are detectable on northern blot at the early bi-nucleate stage and accumulate until the pollen has reached maturity. During germination and pollen tube growth in vitro new NTP303 transcripts appear. This transcription has been proved by northern blots as well as by pulse labelling experiments. Nucleotide sequence analysis revealed that NTPc303 has an open reading frame coding for a predicted protein of 62 kDa. This protein shares homology to ascorbate oxidase and other members of the blue copper oxidase family. A possible function for this clone during pollen germination is discussed.

  11. Effect of TH-lines and clones on the growth and differentiation of B cell clones in microculture.

    Science.gov (United States)

    Kotloff, D B; Cebra, J J

    1988-02-01

    Antibody isotype expression by B cell clones was analyzed using in vitro microcultures containing low numbers of hapten-gelatin-enriched B cells and higher numbers of hemocyanin-specific helper T cell lines or clones. Twenty-eight to sixty-three percent of clones grown in microculture with haptenated hemocyanin and T cells from established lines expressed IgG and/or IgA isotypes in random mixtures, almost always accompanied by IgM. Helper T cells from hemocyanin-specific clones also supported the expression of non-IgM isotypes by the B cell clones, suggesting that a single specificity of T cell can provide sufficient growth and differentiation factors for the display of isotype switching. A positive correlation between the antibody output of clones and the expression of non-IgM isotypes indicated that the switching process may be associated with cell division. Although memory B cells that give clones expressing IgG and/or IgA in the absence of IgM are also enriched on haptenated gelatin, they are not stimulable under conditions of this microculture assay.

  12. Cloning and Expression of Nano Body Gene against Enterotoxin B of Staphylococcus Aureus

    Directory of Open Access Journals (Sweden)

    Zahra Tavassoli

    2017-02-01

    Full Text Available Background & Objectives: Staphylococcus aureus bacteria causes many different diseases by secretion of various enterotoxins. Therefore, it is necessary to develop ways that facilitate the detection of enterotoxins. Nowadays, immunochemical methods which are based on monoclonal antibody technology are used. The heavy chain antibodies that are called VHH or Nano body were found in blood serum of the Camelidae family. The unique properties of this antibody such as their binding to small molecules like toxins make them attractive candidates for the development of immunodiagnostic tests. The present study was done to achieve a VHH molecules against Staphylococcus enterotoxin B. Materials & Methods: Freighting phage library for isolate private Nano bodies against enterotoxin B was done in previous works. Next, pCANTAB 5E vector that consists VHH, extracted from E.coli bacteria strain xl1blue, and after doing PCR process with relative primers, sub cloning in pET21a(+ as an expression vector with cut sites NdeI and XhoI was done. Transformation in E.coli bacteria strain BL21(DE3 was done. Then, the cells effected with IPTG and producing time, and other terms were optimized. Finally, the expression of the protein with SDS-PAGE and western blot techniques was evaluated. Result: For proving cloning of nano body gene in pET21a (+ vector, nucleotide sequence of gene was analyzed, and transforming to E.coli bacteria strain BL21(DE3 was successful. After inspiration, active protein in cell was seen by SDS-PAGE technique and proved by western blot. Conclusion: cloning, sub cloning, and nonabody expression were surveyed in this research. Production of this protein can help to develop new therapeutic methods and produce vaccine against enterotoxin B of Staphylococcus aureus

  13. Cloning, expression, purification and crystallization of dihydrodipicolinate synthase from Agrobacterium tumefaciens

    International Nuclear Information System (INIS)

    Atkinson, Sarah C.; Dogovski, Con; Dobson, Renwick C. J.; Perugini, Matthew A.

    2012-01-01

    Dihydrodipicolinate synthase from the plant pathogen A. tumefaciens has been cloned, expressed, purified and crystallized in its unliganded form, in the presence of its substrate pyruvate and in the presence of pyruvate and the allosteric inhibitor lysine. Diffraction data for the crystals were collected to a maximum resolution of 1.40 Å. Dihydrodipicolinate synthase (DHDPS) catalyzes the first committed step of the lysine-biosynthesis pathway in bacteria, plants and some fungi. This study describes the cloning, expression, purification and crystallization of DHDPS (NP-354047.1) from the plant pathogen Agrobacterium tumefaciens (AgT-DHDPS). Enzyme-kinetics studies demonstrate that AgT-DHDPS possesses DHDPS activity in vitro. Crystals of AgT-DHDPS were grown in the unliganded form and in forms with substrate bound and with substrate plus allosteric inhibitor (lysine) bound. X-ray diffraction data sets were subsequently collected to a maximum resolution of 1.40 Å. Determination of the structure with and without substrate and inhibitor will offer insight into the design of novel pesticide agents

  14. SOME CONSIDERATIONS ON THE PROSPECTS OF SORGHUM CROP

    Directory of Open Access Journals (Sweden)

    Agatha POPESCU

    2014-10-01

    Full Text Available The paper purpose was to analyze the sorghum statement at world, EU and Romania level in order to establish the main trends in the future of this crop. Sorghum is an important cereal coming on the 5th position after maize, rice, wheat and barley at world level due to its importance in human nutrition, animal feed, in producing bioethanol and green energy, and due to its good impact on environment. It is cultivated on all the continents, in the tropical, subtropical and temperate areas due to its resistance to drought, production potential, low inputs and production cost. It is an alternative to maize crop being more utilized as substituent in animal diets. The world sorghum production reached 63,811 thousand metric tons in 2014, the main producers being the USA, Mexico, Nigeria, India, Argentina, Ethiopia, Sudan and China. The world consumption of sorghum reached 63,148 thousand metric tons and it is continuously increasing. The sorghum exports accounted for 7,690 thousand metric tons in 2014, of which the USA export represents 4,600 thousand metric tons. Besides the USA, other exporting countries are Argentina, Australia, Ethiopia, India, Nigeria, Uruguay, while the main importing countries are China, Japan, Chile, Colombia, Mexico, the EU, Sudan. In 2014, the EU produced 576 thousand metric tons sorghum, imported 200 thousand metric tons, and consumed 770 thousand metric tons. The main EU producers of sorghum are France, Italy, Romania, Spain and Hungary. In 2012, Romania cultivated 20,000 ha with sorghum crop, 18 times more than in 2077. Also, in 2012, Romania produced 37.5 thousand tons of sorghum grains, by 31 times more than in 2007. The sorghum yield was 1,875 kg/ha by 66% higher in 2012 compared to 2007. Therefore, these figures show the increasing importance of sorghum crop at world level. Because Romania is situated in suitable geographical area for producing sorghum, it could increase production and become a more important supplier

  15. Fermentation and enzyme treatments for sorghum

    Directory of Open Access Journals (Sweden)

    Patrícia Fernanda Schons

    2012-03-01

    Full Text Available Sorghum (Sorghum bicolor Moench is the fifth most produced cereal worldwide. However, some varieties of this cereal contain antinutritional factors, such as tannins and phytate that may form stable complexes with proteins and minerals which decreases digestibility and nutritional value. The present study sought to diminish antinutritional tannins and phytate present in sorghum grains. Three different treatments were studied for that purpose, using enzymes tannase (945 U/Kg sorghum, phytase (2640 U/Kg sorghum and Paecilomyces variotii (1.6 X 10(7 spores/mL; A Tannase, phytase and Paecilomyces variotii, during 5 and 10 days; B An innovative blend made of tanase and phytase for 5 days followed by a Pv increase for 5 more days; C a third treatment where the reversed order of B was used starting with Pv for 5 days and then the blend of tannase and phytase for 5 more days. The results have shown that on average the three treatments were able to reduce total phenols and both hydrolysable and condensed tannins by 40.6, 38.92 and 58.00 %, respectively. Phytase increased the amount of available inorganic phosphorous, on the average by 78.3 %. The most promising results concerning tannins and phytate decreases were obtained by the enzymes combination of tannase and phytase. The three treatments have shown effective on diminishing tannin and phytate contents in sorghum flour which leads us to affirm that the proposed treatments can be used to increase the nutritive value of sorghum grains destined for either animal feeds or human nutrition.

  16. Fermentation and enzyme treatments for sorghum.

    Science.gov (United States)

    Schons, Patrícia Fernanda; Battestin, Vania; Macedo, Gabriela Alves

    2012-01-01

    Sorghum (Sorghum bicolor Moench) is the fifth most produced cereal worldwide. However, some varieties of this cereal contain antinutritional factors, such as tannins and phytate that may form stable complexes with proteins and minerals which decreases digestibility and nutritional value. The present study sought to diminish antinutritional tannins and phytate present in sorghum grains. Three different treatments were studied for that purpose, using enzymes tannase (945 U/Kg sorghum), phytase (2640 U/Kg sorghum) and Paecilomyces variotii (1.6 X 10(7) spores/mL); A) Tannase, phytase and Paecilomyces variotii, during 5 and 10 days; B) An innovative blend made of tanase and phytase for 5 days followed by a Pv increase for 5 more days; C) a third treatment where the reversed order of B was used starting with Pv for 5 days and then the blend of tannase and phytase for 5 more days. The results have shown that on average the three treatments were able to reduce total phenols and both hydrolysable and condensed tannins by 40.6, 38.92 and 58.00 %, respectively. Phytase increased the amount of available inorganic phosphorous, on the average by 78.3 %. The most promising results concerning tannins and phytate decreases were obtained by the enzymes combination of tannase and phytase. The three treatments have shown effective on diminishing tannin and phytate contents in sorghum flour which leads us to affirm that the proposed treatments can be used to increase the nutritive value of sorghum grains destined for either animal feeds or human nutrition.

  17. Cloning and Expression of 31kDa Outer Membrane Protein of Brucella melitansis in E.coli

    Directory of Open Access Journals (Sweden)

    Sayeneh Khodadadi

    2012-04-01

    Full Text Available Background & Objectives: The identification of Brucella spp. antigens with the capacity to elicit a protective immune response is of the great interest for the researchers. So, characterization and assessment of diverse antigens of Brucella need to be evaluated. In this study, we report the cloning and expression of the gene coding for 31 KDa OMP (OMP31 of Brucella melitensis 16M.   Methods: Brucella melitensis Omp31 gene was amplified with specific primers, cloned into pJET1/2 and subsequently subcloned in pET28a (+ vector. Both these recombinant plasmids were sequenced and then after, expression of recombinant protein was induced by 1mM IPTG. Western blot analysis was also performed by polyclonal rabbit antiserum.   Results: Omp31 successfully was cloned in both plasmid vectors. The recombinant Omp31 was expressed in E.coli host and purified with significant yield. Western blot results along with those of sequencing ensured accurate production of recombinant omp31 and retaining of its partial epitopes.   Conclusion: Our results show that, an expression host such as E. coli is suitable for omp31 production.

  18. Cloning and Expression of Ontak Immunotoxin Using Intein Tag

    Directory of Open Access Journals (Sweden)

    SA Moosavizadeh

    2016-06-01

    Full Text Available Introduction: Inteins (INT are internal parts of a number of proteins in yeast and some other unicellular eukaryotes, which can be separated from the immature protein during protein splicing process. After identifying the mechanism of intein action, applications of these sequences are be considered in the single- step purification of recombinant proteins and different intein tags were developed. The most important advantage of using intein tags in purification of recombinant proteins than other affinity tags is no requirement of expensive protease enzymes and following additional steps to remove protease that make intein tags economically are considered more important. In the present study, denileukin diftitox immunotoxin (brand name Ontak, be fused with an intein tag and it was inserted in pTXB1 plasmid. Methods: In this study, with respect to multiple cloning sites (MCS of pTXB1, specific primers were designed. Polymerase Chain Reaction (PCR was performed and encoding sequence of ONTAK was cloned using restriction sites of NdeI and SapI. Recombinant vector (PTX-IDZ was transformed into E. coli strain ER2566 and expression of gene was studied. Results: The accuracy of recombinant construct was confirmed by PCR and enzymatic digestion. The produced recombinant proteins were confirmed by SDS-PAGE and Western blotting. Conclusion: Restriction site of SapI guarantees no additional residues incorporate in primary protein sequence. Also, the expression of this construct was analyzed in compare with fused protein to poly-His tag. According to the appropriate expression of fused protein in both constructs it was expected that one step- purification of considered drug protein will be success in the following steps.

  19. Hepatocyte specific expression of human cloned genes

    Energy Technology Data Exchange (ETDEWEB)

    Cortese, R

    1986-01-01

    A large number of proteins are specifically synthesized in the hepatocyte. Only the adult liver expresses the complete repertoire of functions which are required at various stages during development. There is therefore a complex series of regulatory mechanisms responsible for the maintenance of the differentiated state and for the developmental and physiological variations in the pattern of gene expression. Human hepatoma cell lines HepG2 and Hep3B display a pattern of gene expression similar to adult and fetal liver, respectively; in contrast, cultured fibroblasts or HeLa cells do not express most of the liver specific genes. They have used these cell lines for transfection experiments with cloned human liver specific genes. DNA segments coding for alpha1-antitrypsin and retinol binding protein (two proteins synthesized both in fetal and adult liver) are expressed in the hepatoma cell lines HepG2 and Hep3B, but not in HeLa cells or fibroblasts. A DNA segment coding for haptoglobin (a protein synthesized only after birth) is only expressed in the hepatoma cell line HepG2 but not in Hep3B nor in non hepatic cell lines. The information for tissue specific expression is located in the 5' flanking region of all three genes. In vivo competition experiments show that these DNA segments bind to a common, apparently limiting, transacting factor. Conventional techniques (Bal deletions, site directed mutagenesis, etc.) have been used to precisely identify the DNA sequences responsible for these effects. The emerging picture is complex: they have identified multiple, separate transcriptional signals, essential for maximal promoter activation and tissue specific expression. Some of these signals show a negative effect on transcription in fibroblast cell lines.

  20. Differential Regulation of Gene Expression of Alveolar Epithelial Cell Markers in Human Lung Adenocarcinoma-Derived A549 Clones

    Directory of Open Access Journals (Sweden)

    Hiroshi Kondo

    2015-01-01

    Full Text Available Stem cell therapy appears to be promising for restoring damaged or irreparable lung tissue. However, establishing a simple and reproducible protocol for preparing lung progenitor populations is difficult because the molecular basis for alveolar epithelial cell differentiation is not fully understood. We investigated an in vitro system to analyze the regulatory mechanisms of alveolus-specific gene expression using a human alveolar epithelial type II (ATII cell line, A549. After cloning A549 subpopulations, each clone was classified into five groups according to cell morphology and marker gene expression. Two clones (B7 and H12 were further analyzed. Under serum-free culture conditions, surfactant protein C (SPC, an ATII marker, was upregulated in both H12 and B7. Aquaporin 5 (AQP5, an ATI marker, was upregulated in H12 and significantly induced in B7. When the RAS/MAPK pathway was inhibited, SPC and thyroid transcription factor-1 (TTF-1 expression levels were enhanced. After treatment with dexamethasone (DEX, 8-bromoadenosine 3′5′-cyclic monophosphate (8-Br-cAMP, 3-isobutyl-1-methylxanthine (IBMX, and keratinocyte growth factor (KGF, surfactant protein B and TTF-1 expression levels were enhanced. We found that A549-derived clones have plasticity in gene expression of alveolar epithelial differentiation markers and could be useful in studying ATII maintenance and differentiation.

  1. Enhanced ethanol production from stalk juice of sweet sorghum by ...

    African Journals Online (AJOL)

    Sweet sorghum (sugar sorghum, Sorghum bicolor) is one kind of non-grain energy crops. As a novel green regenerated high-energy crop with high utility value, high yield of biomass, the sweet sorghum is widely used and developed in China. Stalk juice of sweet sorghum was used as the main substrate for ethanol ...

  2. Effect of heat moisture treatment (HMT) on product quality of sorghum starch

    Science.gov (United States)

    Haryani, Kristinah; Hadiyanto, Handayani, Noera; Nugraheni, Dwi; Suryanto

    2015-12-01

    Sorghum is a cereal plant that rich of nutrition contents. The high content of carbohydrate in sorghum make this plant can be processed into one of the processed food i.e vermicelli. To give better quality, it is necessary to use flour substitution from sorghum starch. The aim of this study was to evaluate the treatment of natural sorghum starch substitution, the addition of CMC, and a comparison of the natural starch with starch sorghum forage sorghum against solid losses value, rehydration weight and texture profiles. The variable used in this study: amount of natural sorghum starch subtituion (10%, 20%, 30%, 40%, 50%), the addition of CMC (0.1%; 0.2%; 0.3%; 0.4%; 0.5%) and substituting sorghum starch Natural: HMT sorghum starch (1: 1; 1: 2; 1: 3; 1: 4; 1: 5) and the quality parameters were evaluated. The result indicated that to substitute sorghum starch naturally at a rate of 50% had the best results with a value of solid losses 5.1% (white sorghum) 5.83% (red sorghum) and weighing rehydration 301.82% (white sorghums) 293.16% (red sorghum), the addition of CMC with 0.5% concentration of 3.96% solid losses value (red sorghum) 4:21% (white sorghums) and weight rehydration 252.71% (white sorghums) 244.45% (red sorghums).

  3. Nutritive value of diferents silage sorghum (Sorghum bicolor L. Moench cultivares - doi: 10.4025/actascianimsci.v34i2.12853

    Directory of Open Access Journals (Sweden)

    Luiz Henrique dos Santos Gomes

    2012-03-01

    Full Text Available Nutrition values of silages from different sorghum cultivars are evaluated. Five 26-kg castrated crossbred lambs, housed in pens equipped with feces and urine collectors for the study of their metabolism, were employed in a 5 x 5 Latin square experimental design. Treatments consisted of silage from five different sorghum cultivars: IPA 1011 and IPA 2564 (grain sorghum, IPA 2502 (dual purpose sorghum, IPA FS-25 and IPA 467 (forage sorghum. Protein level was corrected to 12% by adding a mixture of urea: ammonium sulfate (9:1. Treatments IPA 1011, IPA 2564 and IPA 2502 provided high intake of dry matter, total carbohydrate and total digestible nutrients, and low intake of neutral detergent fiber. Cultivars IPA 1011 and IPA 2564 provided high apparent crude protein digestibility coefficient, whereas cultivars IPA 1011 and IPA 2564 had high total digestible nutrient levels. All cultivars provided positive nitrogen. Owing to nutrient intake and digestibility values, grain sorghum silages evidenced high potential in ruminant nutrition.

  4. Cloning, Expression and Purification of the Recombinant HIV-1 Tat-Nef Fusion Protein in Prokaryotic Expression System

    OpenAIRE

    Somayeh Kadkhodayan; Shiva Irani; Seyed Mehdi Sadat; Fatemeh Fotouhi; Azam Bolhassani

    2016-01-01

    Abstract Background: Nef is one of the HIV-1 critical proteins, because it is essential for viral replication and AIDS disease progression and induction of immune response against it can partially inhibit viral infection. Moreover, a domain of the HIV-1 Trans-Activator of Transcription (Tat, 48-60 aa) could act as a cell penetrating peptide (CPP). In current study, cloning and expression of Tat-Nef fusion protein was performed in E. coli for the first time. The protein expression was confi...

  5. Comparative Analysis of CDPK Family in Maize, Arabidopsis, Rice, and Sorghum Revealed Potential Targets for Drought Tolerance Improvement

    Directory of Open Access Journals (Sweden)

    Shikha Mittal

    2017-12-01

    Full Text Available Calcium dependent protein kinases (CDPKs play significant role in regulation of plant growth and development in response to various stresses including drought. A set of 32 CDPK genes identified in maize were further used for searching of orthologs in the model plant Arabidopsis (72 and major food crops such as rice (78 and sorghum (91. We comprehensively studied the phylogenetic relationship, annotations, gene duplications, gene structure, divergence time, 3-D protein structures and tissue-specific drought induced expression of CDPK genes in all four species. Variation in intron frequency in the studied species was one of the reasons for the functional diversity of CDPK genes to various stress responses. Protein kinase and protein kinase C phosphorylation site domains were the most conserved motifs identified in all species. Four groups were identified from the sequence-based phylogenetic analysis, in which maize CDPKs were clustered in group III. Expression data showed that the CDPK genes were highly expressed in leaf of maize, rice, and sorghum whereas in Arabidopsis the maximum expression was observed in root. The expression assay showed 5, 6, 11, and 9 were the commonly and differentially expressed drought-related orthologous genes in maize, Arabidopsis, rice, and sorghum, respectively. 3-D protein structure were predicted for the nine genes (Arabidopsis: 2, maize: 2, rice: 3, and sorghum: 2 showing differential expression in at least three species. The predicted 3-D structures were further evaluated and validated by Ramachandran plot, ANOLEA, ProSA, and Verify-3D. The superimposed 3-D structure of drought-related orthologous proteins retained similar folding pattern owing to their conserved nature. Functional annotation revealed the involvement of CDPK genes in various pathways such as osmotic homeostasis, cell protection, and root growth. The interactions of CDPK genes in various pathways play crucial role in imparting drought tolerance

  6. Functional cDNA expression cloning: Pushing it to the limit

    Science.gov (United States)

    OKAYAMA, Hiroto

    2012-01-01

    The 1970s and the following decade are the era of the birth and early development of recombinant DNA technologies, which have entirely revolutionized the modern life science by providing tools that enable us to know the structures of genes and genomes and to dissect their components and understand their functions at the molecular and submolecular levels. One major objective of the life sciences is to achieve molecular and chemical understandings of the functions of genes and their encoded proteins, which are responsible for the manifestation of all biological phenomena in organisms. In the early 1980s, I developed, together with Paul Berg, a new technique that enables the cloning of full-length complementary DNAs (cDNAs) on the basis of their functional expression in a given cell of interest. I review the development, application and future implications in the life sciences of this gene-cloning technique. PMID:22450538

  7. CDNA cloning, characterization and expression of an endosperm-specific barley peroxidase

    DEFF Research Database (Denmark)

    Rasmussen, Søren Kjærsgård; Welinder, K.G.; Hejgaard, J.

    1991-01-01

    A barley peroxidase (BP 1) of pI ca. 8.5 and M(r) 37000 has been purified from mature barley grains. Using antibodies towards peroxidase BP 1, a cDNA clone (pcR7) was isolated from cDNA expression library. The nucleotide sequence of pcR7 gave a derived amino acid sequence identical to the 158 C...

  8. Modification of Sorghum Starch-Cellulose Bioplastic with Sorghum Stalks Filler

    Directory of Open Access Journals (Sweden)

    Yuli Darni

    2017-05-01

    Full Text Available This study evaluated the feasibility of bioplastics production by various ratio of sorghum starch and cellulose from red seaweed Eucheuma spinossum, and the use of glycerol as plasticizer and sorghum stalks as filler. Solid-liquid matrix transition should be far over the operating temperature of gelatinization and extracted at 95oC in order to avoid the loss of conductivity. The analyzed variables were starch and cellulose seaweed Eucheuma spinossum and the addition of variation of filler. Sorghum stalk could be expected to affect the mechanical and physical properties of bioplastics. A thin sheet of plastic (plastic film was obtained as a result that have been tested mechanically to obtain the best condition for the formulation of starch-cellulose 8.5:1.5 (g/g. From the result of morphological studies, the fillers in the mixture composites were more randomly in each product and the addition of filler can increase mechanical properties of bioplastics. Chemical modification had a major effect on the mechanical properties. The phenomena of degradation and thermoplasticization were visible at chemical changes that can be observed in FTIR spectrum test results.

  9. Cloning and Expression of a Cytosolic HSP90 Gene in Chlorella vulgaris

    Directory of Open Access Journals (Sweden)

    Zhengyi Liu

    2014-01-01

    Full Text Available Heat shock protein 90 (HSP90, a highly conserved molecular chaperone, plays essential roles in folding, keeping structural integrity, and regulating the subset of cytosolic proteins. We cloned the cDNA of Chlorella vulgaris HSP90 (named CvHSP90 by combining homology cloning with rapid amplification of cDNA ends (RACE. Sequence analysis indicated that CvHSP90 is a cytosolic member of the HSP90 family. Quantitative RT-PCR was applied to determine the expression level of messenger RNA (mRNA in CvHSP90 under different stress conditions. C. vulgaris was kept in different temperatures (5–45°C for 1 h. The mRNA expression level of CvHSP90 increased with temperature from 5 to 10°C, went further from 35 to 40°C, and reached the maximum at 40°C. On the other hand, for C. vulgaris kept at 35°C for different durations, the mRNA expression level of CvHSP90 increased gradually and reached the peak at 7 h and then declined progressively. In addition, the expression level of CvHSP90 at 40 or 45 in salinity (‰ was almost fourfold of that at 25 in salinity (‰ for 2 h. Therefore, CvHSP90 may be a potential biomarker to monitor environment changes.

  10. Cloning and expression of a rat brain α2B-adrenergic receptor

    International Nuclear Information System (INIS)

    Flordellis, C.S.; Handy, D.E.; Bresnahan, M.R.; Zannis, V.I.; Gavras, H.

    1991-01-01

    The authors isolated a cDNA clone (RBα 2B ) and its homologous gene (GRα 2B ) encoding an α 2B -adrenergic receptor subtype by screening a rat brain cDNA and a rat genomic library. Nucleotide sequence analysis showed that both clones code for a protein of 458 amino acids, which is 87% homologous to the human kidney glycosylated adrenergic receptor (α 2 -C4) and divergent from the rat kidney nonglycosylated α 2B subtype (RNGα 2 ). Transient expression of RBα 2B in COS-7 cells resulted in high-affinity saturable binding for [ 3 H]rauwolscine and a high receptor number in the membranes of transfected COS-7 cells. Pharmacological analysis demonstrated that the expressed receptor bound adrenergic ligands with the following order of potency: rauwolscine > yohimbine > prazosin > oxymetazoline, with a prazosin-to-oxymetazoline K i ratio of 0.34. This profile is characteristic of the α 2B -adrenergic receptor subtype. Blotting analysis of rat brain mRNA gave one major and two minor mRNA species, and hybridization with strand-specific probes showed that both DNA strands of GRα 2B may be transcriptionally active. These findings show that rat brain expresses an α 2B -adrenergic receptor subtype that is structurally different from the rat kidney nonglycosylated α 2B subtype. Thus the rat expresses at least two divergent α 2B -adrenergic receptors

  11. Automated cloning methods.; TOPICAL

    International Nuclear Information System (INIS)

    Collart, F.

    2001-01-01

    Argonne has developed a series of automated protocols to generate bacterial expression clones by using a robotic system designed to be used in procedures associated with molecular biology. The system provides plate storage, temperature control from 4 to 37 C at various locations, and Biomek and Multimek pipetting stations. The automated system consists of a robot that transports sources from the active station on the automation system. Protocols for the automated generation of bacterial expression clones can be grouped into three categories (Figure 1). Fragment generation protocols are initiated on day one of the expression cloning procedure and encompass those protocols involved in generating purified coding region (PCR)

  12. Maturation curves of sweet sorghum genotypes

    Directory of Open Access Journals (Sweden)

    Renan Silva e Souza

    2016-02-01

    Full Text Available ABSTRACT Sweet sorghum [Sorghum bicolor (L. Moench] stands out as a complementary crop to sugarcane Saccharum spp. for the production of ethanol, since it has juicy stems with directly fermentable sugars. Due to this fact, there is a need for the analysis of sweet sorghum properties in order to meet the agro-industry demand. This work aimed to develop and study the maturation curves of seven sweet sorghum cultivars in ten harvest dates. The results showed a significant difference between cultivars and harvest dates for all parameters analysed (p≤0.01. Regarding the sugar content, the cultivars BRS508, XBWS80147 and CMSX629 showed the highest means for the total reducing sugars (TRS and recoverable sugar (RS. In the production of ethanol per tonne of biomass (EP, the cultivars BRS508 and CMSX629 presented the best results.

  13. SILAGE QUALITY OF CORN AND SORGHUM ADDED WITH FORAGE PEANUTS

    Directory of Open Access Journals (Sweden)

    WALKÍRIA GUIMARÃES CARVALHO

    2016-01-01

    Full Text Available Corn and sorghum are standard silage crops because of their fermentative characteristics. While corn and sorghum silages have lower crude protein (CP contents than other crops, intercropping with legumes can increase CP content. Furthermore, one way to increase CP content is the addition of legumes to silage. Consequently, the research objective was to evaluate the fermentative and bromatological characteristics of corn (Zea mays and sorghum (Sorghum bicolor silages added with forage peanuts (Arachis pintoi. The experimental design was completely randomized with four replicates. The treatments consisted of corn silage, sorghum silage, forage peanut silage, corn silage with 30% forage peanut, and sorghum silage with 30% forage peanut. The results showed that the corn and sorghum added with peanut helped to improve the silage fermentative and bromatological characteristics, proving to be an efficient technique for silage quality. The forage peanut silage had lower fermentative characteristics than the corn and sorghum silages. However, the forage peanut silage had a greater CP content, which increased the protein contents of the corn and sorghum silages when intercropped with forage peanuts.

  14. [Gene clone and expression of Barx1 in different tooth of the mini-pig at embryonic day 40].

    Science.gov (United States)

    Zhang, Ying; Yin, Ji-rong; Yang, Kai

    2012-10-01

    To partially clone and compare the quantitative expression of tooth development-related gene Barx1 in different teeth of the mini-pig embryo at embryonic day 40, and to investigate the relationship between Barx1 spatial quantitative expression and tooth morphogenesis. The mini-pig Barx1 genes was partially cloned and the mRNA sequences of human Barx1 genes was aligned with expressed sequence tags (EST) of pig by basic local alignment search tool (BLAST), which were assembled with DNAman v5.2.2. With designed primers, Barx1 was partially cloned in use of reverse transcription polymerase chain reaction (PCR), and tested by BLAST with all the species in NCBI database and confirmed as one part of target gene. Laser capture microdissection was used to collect tooth samples from frozen sections which were prepared before in -80°C freezer. Real-time PCR was carried out to analyze quantitative expression in different teeth. Partial mini-pig Barx1 gene of 698 bp was cloned. Real-time PCR showed that, glyceraldehyde-3-phosphate dehydrogenase used as loading control, the figures of 2(-ΔCT) of lower deciduous incisor, canine, the third premolar and molar were 0.000 249, 0.000 715, 0.026 096 and 0.112 656, respectively. There was a trend of increasing expression from anterior to posterior teeth. Barx1 gene could be related to the number or differentiation of tooth cusps.

  15. Mapping and characterisation of the sorghum cell suspension ...

    African Journals Online (AJOL)

    Here we reported the first secretomic study of sorghum (Sorghum bicolor), a naturally drought tolerant cereal crop. In this study, we used a gel-based proteomic approach in combination with mass spectrometry to separate and identify proteins secreted into the culture medium of sorghum cell suspensions, a first step ...

  16. Analysis of nuclear reprogramming in cloned miniature pig embryos by expression of Oct-4 and Oct-4 related genes

    International Nuclear Information System (INIS)

    Lee, Eugine; Lee, So Hyun; Kim, Sue

    2006-01-01

    Xenotransplantation is a rapidly expanding field of research and cloned miniature pigs have been considered as a model animal for it. However, the efficiency of somatic cell nuclear transfer (SCNT) is extremely low, with most clones resulting in early lethality and several kinds of aberrant development. A possible explanation for the developmental failure of SCNT embryos is insufficient reprogramming of the somatic cell nucleus by the oocyte. In order to test this, we analyzed the reprogramming capacity of differentiated fibroblast cell nuclei and embryonic germ cell nuclei with Oct-4 and Oct-4 related genes (Ndp5211, Dppa2, Dppa3, and Dppa5), which are important for embryonic development, Hand1 and GATA-4, which are important for placental development, as molecular markers using RT-PCR. The Oct-4 expression level was significantly lower (P < 0.05) in cloned hatched blastocysts derived from fibroblasts and many of fibroblast-derived clones failed to reactivate at least one of the tested genes, while most of the germ cell clones and control embryos correctly expressed these genes. In conclusion, our results suggest that the reprogramming of fibroblast-derived cloned embryos is highly aberrant and this improper reprogramming could be one reason of the early lethality and post-implantation anomalies of somatic cell-derived clones

  17. Insights into Alpha-Hemolysin (Hla) Evolution and Expression among Staphylococcus aureus Clones with Hospital and Community Origin

    DEFF Research Database (Denmark)

    Tavares, Ana; Nielsen, Jesper B; Boye, Kit

    2014-01-01

    BACKGROUND: Alpha-hemolysin (Hla) is a major virulence factor in the pathogenesis of Staphylococcus aureus infection, being active against a wide range of host cells. Although hla is ubiquitous in S. aureus, its genetic diversity and variation in expression in different genetic backgrounds...... and SCCmec typing. The internal regions of hla and the hla promoter were sequenced and gene expression was assessed by RT-PCR. RESULTS: Alpha-hemolysin encoding- and promoter sequences were diverse, with 12 and 23 different alleles, respectively. Based on phylogenetic analysis, we suggest that hla may have...... in the RNAIII binding site were not associated to hla expression. Although expression rates of hla were in general strain-specific, we observed CA clones showed significantly higher hla expression (p = 0.003) when compared with HA clones. CONCLUSION: We propose that the hla gene has evolved together...

  18. Sweet Sorghum Crop. Effect of the Compost Application

    International Nuclear Information System (INIS)

    Negro, M. J.; Solano, M. L.; Carrasco, J.; Ciria, P.

    1998-01-01

    A 3 year-plot experiments were performed to determined the possible persistence of the positive effects of treating soil with compost. For this purpose, a sweet sorghum bagasse compost has been used. Experiments were achieved with sweet sorghum (Sorghum bicolor. L. Moench) vr Dale as energy crop. Similar sorghum productivities were obtained both in plots with consecutive compost applications and in plots amended with mineral fertilizers. No residual effect after three years has been detected. It could be due to the low dose of compost application. (Author) 27 refs

  19. Expressão fenotípica de clones de seringueira na região noroeste do estado de São Paulo Phenotypic expression of rubber tree clones in the northwestern region of São Paulo state

    Directory of Open Access Journals (Sweden)

    Paulo de Souza Gonçalves

    2006-01-01

    Full Text Available O desenvolvimento de novos clones de seringueira [Hevea brasiliensis (Willd. ex Adr. de Juss. Muell.-Arg.] com alto potencial de produção aliado a outros caracteres secundários desejáveis é de fundamental importância para uma heveicultura sustentável e competitiva. O objetivo deste trabalho foi avaliar a expressão fenotípica de caracteres superiores em 17 clones de seringueira, tendo em vista a escolha dos mais promissores. Em campo, o experimento obedeceu ao delineamento de blocos ao acaso com três repetições e parcelas lineares de seis plantas. Pelos resultados, verificou-se que o clone IAC 40 foi o mais produtivo, com média de 2.316 kg de borracha seca ha-1 ano-1 no período de seis anos, seguido pelo clone IAC 300 (1.921 kg, enquanto o clone-testemunha, RRIM 600 produziu 1.493 kg. Observou-se na maior parte dos clones, crescimento superior em relação à testemunha. A porcentagem de plantas aptas à sangria variou de 40% (IAC 329 a 100% (IAC 327. Exceto nos clones IAC 56, IAC 331 e IAN 3156 com 7,21 mm, 7,18 mm e 6,40 mm respectivamente, em todos os demais notou-se espessura de casca virgem inferior ao clone RRIM 600 (6,38 mm. Com exceção do IAN 3156, os demais clones tiveram baixa incidência de secamento de painel. O bom desempenho de todos os clones IAC e amazônicos (IAN, Fx e RO permite que sejam recomendados para plantio em pequena escala, ao tempo em que serão avaliados para futura recomendação em grande escala envolvendo diferentes ambientes do Estado de São Paulo.The development of new clones with high production combined to other desirable secondary characters is fundamental for a sustainable and competitive rubber tree cultivation. The objective of this study was to evaluate, during a period of 13 years, the phenotypic expression of superior characters of 17 clones of rubber tree grown in the plateau region of São Paulo State, Brazil. The treatments were arranged in a randomized block design with three

  20. Novel storage technologies for raw and clarified syrup biomass feedstocks from sweet sorghum (Sorghum bicolor L. Moench)

    Science.gov (United States)

    Attention is currently focused on developing sustainable supply chains of sugar feedstocks for new, flexible biorefineries. Fundamental processing needs identified by industry for the large-scale manufacture of biofuels and bioproducts from sweet sorghum (Sorghum bicolor L. Moench) include stabiliz...

  1. Identification of Dw1, a Regulator of Sorghum Stem Internode Length.

    Directory of Open Access Journals (Sweden)

    Josie Hilley

    Full Text Available Sorghum is an important C4 grain and grass crop used for food, feed, forage, sugar, and biofuels. In its native Africa, sorghum landraces often grow to approximately 3-4 meters in height. Following introduction into the U.S., shorter, early flowering varieties were identified and used for production of grain. Quinby and Karper identified allelic variation at four loci designated Dw1-Dw4 that regulated plant height by altering the length of stem internodes. The current study used a map-based cloning strategy to identify the gene corresponding to Dw1. Hegari (Dw1dw2Dw3dw4 and 80M (dw1dw2Dw3dw4 were crossed and F2 and HIF derived populations used for QTL mapping. Genetic analysis identified four QTL for internode length in this population, Dw1 on SBI-09, Dw2 on SBI-06, and QTL located on SBI-01 and SBI-07. The QTL on SBI-07 was ~3 Mbp upstream of Dw3 and interacted with Dw1. Dw1 was also found to contribute to the variation in stem weight in the population. Dw1 was fine mapped to an interval of ~33 kbp using HIFs segregating only for Dw1. A polymorphism in an exon of Sobic.009G229800 created a stop codon that truncated the encoded protein in 80M (dw1. This polymorphism was not present in Hegari (Dw1 and no other polymorphisms in the delimited Dw1 locus altered coding regions. The recessive dw1 allele found in 80M was traced to Dwarf Yellow Milo, the progenitor of grain sorghum genotypes identified as dw1. Dw1 encodes a putative membrane protein of unknown function that is highly conserved in plants.

  2. Morphological responses of forage sorghums to salinity and ...

    African Journals Online (AJOL)

    The response of forage sorghum [Sorghum bicolor (L.) Moench] varieties to salinity and irrigation frequency were studied from December 2007 to December 2009. Two forage sorghum varieties (Speedfeed and KFS4) were grown under salinity levels of 0, 5, 10 and 15 dS m-1 and irrigated when the leaf water potential ...

  3. The Sorghum bicolor genome and the diversification of grasses

    Energy Technology Data Exchange (ETDEWEB)

    Paterson, Andrew H.; Bowers, John E.; Bruggmann, Remy; dubchak, Inna; Grimwood, Jane; Gundlach, Heidrun; Haberer, Georg; Hellsten, Uffe; Mitros, Therese; Poliakov, Alexander; Schmutz, Jeremy; Spannagl, Manuel; Tang, Haibo; Wang, Xiyin; Wicker, Thomas; Bharti, Arvind K.; Chapman, Jarrod; Feltus, F. Alex; Gowik, Udo; Grigoriev, Igor V.; Lyons, Eric; Maher, Christopher A.; Martis, Mihaela; Marechania, Apurva; Otillar, Robert P.; Penning, Bryan W.; Salamov, Asaf. A.; Wang, Yu; Zhang, Lifang; Carpita, Nicholas C.; Freeling, Michael; Gingle, Alan R.; hash, C. Thomas; Keller, Beat; Klein, Patricia; Kresovich, Stephen; McCann, Maureen C.; Ming, Ray; Peterson, Daniel G.; ur-Rahman, Mehboob-; Ware, Doreen; Westhoff, Peter; Mayer, Klaus F. X.; Messing, Joachim; Rokhsar, Daniel S.

    2008-08-20

    Sorghum, an African grass related to sugar cane and maize, is grown for food, feed, fibre and fuel. We present an initial analysis of the approx730-megabase Sorghum bicolor (L.) Moench genome, placing approx98percent of genes in their chromosomal context using whole-genome shotgun sequence validated by genetic, physical and syntenic information. Genetic recombination is largely confined to about one-third of the sorghum genome with gene order and density similar to those of rice. Retrotransposon accumulation in recombinationally recalcitrant heterochromatin explains the approx75percent larger genome size of sorghum compared with rice. Although gene and repetitive DNA distributions have been preserved since palaeopolyploidization approx70 million years ago, most duplicated gene sets lost one member before the sorghum rice divergence. Concerted evolution makes one duplicated chromosomal segment appear to be only a few million years old. About 24percent of genes are grass-specific and 7percent are sorghum-specific. Recent gene and microRNA duplications may contribute to sorghum's drought tolerance.

  4. Agronomic and morphological performance of sorghum (sorghum ...

    African Journals Online (AJOL)

    SARAH

    2013-03-30

    Mar 30, 2013 ... occasional frost limits growth and seed set of un- adapted cultivars (Arkel, 1979) making seed multiplication of un-adapted varieties unsuccessful. Previous studies have shown that sorghum cultivars adapted to high altitude, low rainfall areas. Journal of Applied Biosciences 63: 4720 – 4726. ISSN 1997– ...

  5. Cloning and expression of chaetomium thermophilum xylanase 11-A gene in prokaryote

    International Nuclear Information System (INIS)

    Wajid, S.; Latif, F.; Afzal, S.; Rajoka, I.

    2008-01-01

    The xylanase gene was cloned into pET32a(+) and expressed in E. coli BL21 under T7 promotor alongwith fusion protein. The SDS-PAGE and western blot analysis showed a protein of 42 kDa. The best expression of xylanase enzyme was found by using xylose as carbon source and lactose as an inducer. The maximum activity of xylanase expressed in E. coli was 6.02 U/mL in the presence of 2% xylose in DS medium. The activity of recombinant xylanase was observed on 1% xylan LB agar plates, showed halos of xylan clearance when lactose was used as an inducer. (author)

  6. Assessment of Genetic Variability in Sorghum Accessions (Sorghum ...

    African Journals Online (AJOL)

    ADOWIE PERE

    The polymorphic information content (PIC) of individual primer ranged from 0.34 to 0.70 with a mean value of 0.54 indicating enough ... Keywords: Sorghum; Simple Sequence Repeat markers; Genetic variation; Polymorphic Information Content;. Coefficient of ... based techniques include Restriction Fragment Length.

  7. Productivity and Competitiveness of Sorghum Production in ...

    African Journals Online (AJOL)

    showed that sorghum production in the study areas yielded profitable returns ... Keywords: Sorghum, Profitability, Competitiveness, Investment Potential, .... Guinness Ghana Brewery Limited to estimate cost and returns at the marketing sector ...

  8. A sorghum (Sorghum bicolor mutant with altered carbon isotope ratio.

    Directory of Open Access Journals (Sweden)

    Govinda Rizal

    Full Text Available Recent efforts to engineer C4 photosynthetic traits into C3 plants such as rice demand an understanding of the genetic elements that enable C4 plants to outperform C3 plants. As a part of the C4 Rice Consortium's efforts to identify genes needed to support C4 photosynthesis, EMS mutagenized sorghum populations were generated and screened to identify genes that cause a loss of C4 function. Stable carbon isotope ratio (δ13C of leaf dry matter has been used to distinguishspecies with C3 and C4 photosynthetic pathways. Here, we report the identification of a sorghum (Sorghum bicolor mutant with a low δ13C characteristic. A mutant (named Mut33 with a pale phenotype and stunted growth was identified from an EMS treated sorghum M2 population. The stable carbon isotope analysis of the mutants showed a decrease of 13C uptake capacity. The noise of random mutation was reduced by crossing the mutant and its wildtype (WT. The back-cross (BC1F1 progenies were like the WT parent in terms of 13C values and plant phenotypes. All the BC1F2 plants with low δ13C died before they produced their 6th leaf. Gas exchange measurements of the low δ13C sorghum mutants showed a higher CO2 compensation point (25.24 μmol CO2.mol-1air and the maximum rate of photosynthesis was less than 5μmol.m-2.s-1. To identify the genetic determinant of this trait, four DNA pools were isolated; two each from normal and low δ13C BC1F2 mutant plants. These were sequenced using an Illumina platform. Comparison of allele frequency of the single nucleotide polymorphisms (SNPs between the pools with contrasting phenotype showed that a locus in Chromosome 10 between 57,941,104 and 59,985,708 bps had an allele frequency of 1. There were 211 mutations and 37 genes in the locus, out of which mutations in 9 genes showed non-synonymous changes. This finding is expected to contribute to future research on the identification of the causal factor differentiating C4 from C3 species that can be used

  9. Development of SSR Markers Linked to Low Hydrocyanic Acid Content in Sorghum-Sudan Grass Hybrid Based on BSA Method.

    Science.gov (United States)

    Xiao-Xia, Yu; Zhi-Hua, Liu; Zhuo, Yu; Yue, Shi; Xiao-Yu, Li

    2016-01-01

    Sorghum-Sudan grass hybrid containing high hydrocyanic acid content can cause hydrocyanic acid poisoning to the livestock and limit the popularization of this forage crop. Molecular markers associated with low hydrocyanic acid content can speed up the process of identification of genotypes with low hydrocyanic acid content. In the present study, 11 polymorphic SSR primers were screened and used for bulked segregant analysis and single marker analysis. Three SSR markers Xtxp7230, Xtxp7375 and Bnlg667960 associated with low hydrocyanic acid content were rapidly identified by BSA. In single marker analysis, six markers Xtxp7230, Xtxp7375, Bnlg667960, Xtxp67-11, Xtxp295-7 and Xtxp12-9 were linked to low hydrocyanic acid content, which explained the proportion of phenotypic variation from 7.6 % to 41.2 %. The markers identified by BSA were also verified by single marker analysis. The three SSR marker bands were then cloned and sequenced for sequence homology analysis in NCBI. It is the first report on the development of molecular markers associated with low hydrocyanic acid content in sorghum- Sudan grass hybrid. These markers will be useful for genetic improvement of low hydrocyanic acid sorghum-Sudan grass hybrid by marker-assisted breeding.

  10. Molecular Cloning, Expression and Characterization of Plasmid Encoding Rhomboid 4 (ROM4 of Tachyzoite of Toxoplasma gondii RH Strain

    Directory of Open Access Journals (Sweden)

    Mohammad Taghi RAHIMI

    2017-12-01

    Full Text Available AbstractBackground: The objective of this study was to clone, express and characterize the gene encoding rhomboid 4 (ROM4 proteins, a vital gene in surface adhesion and host cell invasion process of tachyzoite of T. gondii in an appropriate expression vector and eukaryotic cell for production of recombinant protein.Methods: Toxoplasma RNA was isolated from tachyzoites (RH strain and complementary DNA was synthesized. Oligonucleotide primer pair was designed based on Toxoplasma ROM4 gene sequence with XhoI and EcoRI restriction sites at 5´ end of forward and reverse primers, respectively. ROM4 gene was amplified by PCR, cloned into pTG19-T vector and the recombinant plasmid was sequenced. The gene was subcloned into pcDNA3 plasmid and expressed in CHO cells as eukaryotic cell. SDS-PAGE and western blotting were performed for protein determination and verification.Results: Cloning of ROM4 gene in pTG19-T vector was confirmed by colony-PCR and enzymatic digestion. The results of enzymatic digestion and gene sequencing confirmed successful cloning and subcloning procedures. The nucleotide sequence of the cloned ROM4 gene showed 99% homology compared to the corresponding sequences of original gene. SDS-PAGE and western blotting analyses of the purified protein revealed a single band having expected size of 65 kDa.Conclusion: This eukaryotic expression system is an appropriate system for high-level recombinant protein production of ROM4 gene from T. gondii tachyzoites used as antigenic component for serological assay and vaccine development.

  11. The productive potentials of sweet sorghum ethanol in China

    International Nuclear Information System (INIS)

    Zhang, Caixia; Xie, Gaodi; Li, Shimei; Ge, Liqiang; He, Tingting

    2010-01-01

    As one of the important non-grain energy crops, sweet sorghum has attracted the attention of scientific community and decision makers of the world since decades. But insufficient study has been done about the spatial suitability distribution and ethanol potential of sweet sorghum in China. This paper attempts to probe into the spatial distribution and ethanol potential of sweet sorghum in China by ArcGIS methods. Data used for the analysis include the spatial data of climate, soil, topography and land use, and literatures relevant for sweet sorghum studies. The results show that although sweet sorghum can be planted in the majority of lands in China, the suitable unused lands for large-scale planting (unit area not less than 100 hm 2 ) are only as much as 78.6 x 10 4 hm 2 ; and the productive potentials of ethanol from these lands are 157.1 x 10 4 -294.6 x 10 4 t/year, which can only meet 24.8-46.4% of current demand for E10 (gasoline mixed with 10% ethanol) in China (assumption of the energy efficiency of E10 is equivalent to that of pure petroleum). If all the common grain sorghum at present were replaced by sweet sorghum, the average ethanol yield of 244.0 x 10 4 t/year can be added, and thus the productive potentials of sweet sorghum ethanol can satisfy 63.2-84.9% of current demand for E10 of China. In general, Heilongjiang, Jilin, Inner Mongolia and Liaoning rank the highest in productive potentials of sweet sorghum ethanol, followed by Hebei, Shanxi, Sichuan, and some other provinces. It is suggested that these regions should be regarded as the priority development zones for sweet sorghum ethanol in China.

  12. Genetic diversity in sorghum transpiration efficiency

    Science.gov (United States)

    Sorghum is the fifth most important grain crop and is becoming increasingly important as a biofuel feedstock due to its superior tolerance to water deficit stress. Sorghum is commonly grown under rain-fed conditions in the Southern Plains and other semi-arid regions in the world. Thus, its product...

  13. Chemical control of wild sorghum (sorghum arundinaceum Del. Stapf. in faba bean (vicia faba L.) in the Northern State of Sudan

    International Nuclear Information System (INIS)

    Bedry, K. A. M.; Elamin, A. E. M.

    2011-01-01

    An experiment was conducted at Merowe Research Station farm, in the Northern State, Sudan, during 2008/2009 and 2009/2010 seasons. The objectives of the experiment were to determine the damage inflicted by a wild sorghum species (Sorghum arundinaceum (Del.) Stapf. ) on the yield of faba bean (Vicia faba L.) and to evaluate the efficacy of the post-emergence herbicide clodinafop-propargyl (Topik) on wild sorghum and its effect on faba bean yield. The wild sorghum reduced faba bean crop stand and straw and seed yields by 53% - 76%, 76% - 79% and 88% - 91%, respectively, compared with the hand-weeded control. Faba bean was tolerant to the herbicide. The herbicide, at all rates, effected complete (100%) and persistent control of the wild sorghum and resulted in faba bean seed yield comparable to the hand-weeded control. The lowest dose (0.075 kg a.i/ha) of the herbicide used was equal to 75% of the dose recommended for the control of wild sorghum in wheat. It is concluded that clodinafop-propargyl at 0.075 kg a.e/ha could be used in controlling wild sorghum in faba bean. At this rate, the marginal rate of return was about 35 which indicating that every monetary unit (SDG 1) invested in the mentioned treatment would be returned back, plus additional amount of 35 SDG.(Author)

  14. Cloning and expression study of BnaLCR78 in Brassica napus

    International Nuclear Information System (INIS)

    Zhuang, L.; Ze, L. Y.; Cheng, W. Y.

    2016-01-01

    BnaLCR78 genes of three types of rape were cloned in rape (Brassica napus), and encoded protein structure was analyzed, the Results showed that the protein had a conserved coding domain which was analogues among LCR family of Arabidopsis. The expression patterns of genes of three types of rape in varying tissues and in specific same tissues were analyzed using quantitative method. The Results showed that their expression patterns differ from that of former research in Brassica napus, which may result from the difference of sampling time. We speculated that the gene might be involved in transpiration and transportation and distribution of nutrient, oil content in seed. (author)

  15. Molecular cloning and expression of bovine kappa-casein in Escherichia coli

    International Nuclear Information System (INIS)

    Kang, Y.C.; Richardson, T.

    1988-01-01

    A cDNA library was constructed using poly(A) + RNA from bovine mammary gland. This cDNA library of 6000 clones was screened employing colony hybridization using 32 P-labelled oligonucleotide probes and restriction endonuclease mapping. The cDNA from the selected plasmid, pKR76, was sequenced using the dideoxy-chain termination method. The cDNA insert of pKR76 carries the full-length sequence, which codes for mature kappa-casein protein. The amino acid sequence deduced from the cDNA sequence fits the published amino acid sequence with three exceptions; the reported pyroglutamic acid at position 1, tyrosine at position 35, and aspartic acid at position 81 are, respectively, a glutamine, a histidine, and an asparagine in the clone containing pKR76. The MspI-, NlaIV-cleaved fragment (630 base pair) from the kappa-casein cDNA insert has been subcloned into expression vectors pUC18 and pKK233-2, which contain a lac promoter and a trc promoter, respectively. Escherichia coli cells carrying the recombinant expression plasmids were shown to produce kappa-casein protein having the expected mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and being recognized by specific antibodies raised against natural bovine kappa-casein

  16. Cloning and expression of chaetomium thermophilum xylanase 11-A

    International Nuclear Information System (INIS)

    Andleeb, S.; Latif, F.; Afzal, S.; Mukhtar, Z.; Mansoor, S.; Rajoka, I.

    2008-01-01

    The various thermophilic fungi like Chaetomium thermophile has potential to secrete xylanase and cellulase enzymes. In the present study eukaryotic expression system of Pichia pastoris (yeast) was used to express xylanase gene. The xylanase (Xyn 11-A) gene was isolated from C. thermophile strain NIBGE-1. Primers were designed to amplify the gene, ligated into P. pastoris pPIC3.5K vector, the resultant recombinant clone pSSZ810 was transformed into the genome of P. pastoris GS115 strain through electroporation. Transformants were selected on yeast peptone dextrose medium (YPD) plates containing antibiotic geneticin (100 mg/ml) upto final concentration of 0.75 mg/ml. The maximum activity of xylanase 2.04 U/ml after incubation of 2 hours at 50 degree C was observed in the presence of 100% methanol inducer upto final concentration of 30 macro L (0.5%) as compared to control. HPLC analysis represented high peak of xylose as compared to control. SDS-PAGE indicated approx. 28 kDa protein of expressed xylanase gene. (author)

  17. Cloning and expression of the human N-methyl-D-aspartate receptor subunit NR3A

    DEFF Research Database (Denmark)

    Eriksson, Maria; Nilsson, Anna; Froelich-Fabre, Susanne

    2002-01-01

    Native N-methyl-D-aspartate (NMDA) receptors are heteromeric assemblies of four or five subunits. The NMDA receptor subunits, NR1, NR2A, NR2B, NR2C, and NR2D have been cloned in several species, including man. The NR3A subunit, which in rodents is predominantly expressed during early development......, seems to function by reducing the NMDA receptor response. The human homologue to the rat NR3A, however, had not been cloned. In order to study the functions of the human NR3A (hNR3A), we have cloned and sequenced the hNR3A. It was found to share 88% of the DNA sequence with the rat gene, corresponding...

  18. 7339 BASELINE SURVEY ON FACTORS AFFECTING SORGHUM ...

    African Journals Online (AJOL)

    muuicathy

    2013-01-01

    Jan 1, 2013 ... factors affecting sorghum production and the sorghum farming ... The informal seed system includes methods such as retaining seed on-farm from ..... Jaetzold R and H Schmidt Farm Management Handbook of Kenya, Ministry.

  19. Effects of Sorghum [Sorghum bicolor (L. Moench] Crude Extracts on Starch Digestibility, Estimated Glycemic Index (EGI, and Resistant Starch (RS Contents of Porridges

    Directory of Open Access Journals (Sweden)

    Dilek Lemlioglu-Austin

    2012-09-01

    Full Text Available Bran extracts (70% aqueous acetone of specialty sorghum varieties (tannin, black, and black with tannin were used to investigate the effects of sorghum phenolic compounds on starch digestibility, Estimated Glycemic Index (EGI, and Resistant Starch (RS of porridges made with normal corn starch, enzyme resistant high amylose corn starch, and ground whole sorghum flours. Porridges were cooked with bran extracts in a Rapid Visco-analyser (RVA. The cooking trials indicated that bran extracts of phenolic-rich sorghum varieties significantly reduced EGI, and increased RS contents of porridges. Thus, there could be potential health benefits associated with the incorporation of phenolic-rich sorghum bran extracts into foods to slow starch digestion and increase RS content.

  20. Cloning and molecular analyses of a gibberellin 20-oxidase gene expressed specifically in developing seeds of watermelon.

    Science.gov (United States)

    Kang, H G; Jun, S H; Kim, J; Kawaide, H; Kamiya, Y; An, G

    1999-10-01

    To understand the biosynthesis and functional role of gibberellins (GAs) in developing seeds, we isolated Cv20ox, a cDNA clone from watermelon (Citrullus lanatus) that shows significant amino acid homology with GA 20-oxidases. The complementary DNA clone was expressed in Escherichia coli as a fusion protein, which oxidized GA(12) at C-20 to the C(19) compound GA(9), a precursor of bioactive GAs. RNA-blot analysis showed that the Cv20ox gene was expressed specifically in developing seeds. The gene was strongly expressed in the integument tissues, and it was also expressed weakly in inner seed tissues. In parthenocarpic fruits induced by 1-(2-chloro-4-pyridyl)-3-phenylurea treatment, the expression pattern of Cv20ox did not change, indicating that the GA 20-oxidase gene is expressed primarily in the maternal cells of developing seeds. The promoter of Cv20ox was isolated and fused to the beta-glucuronidase (GUS) gene. In a transient expression system, beta-glucuronidase staining was detectable only in the integument tissues of developing watermelon seeds.

  1. Genetic architecture of kernel composition in global sorghum germplasm

    Science.gov (United States)

    Sorghum [Sorghum bicolor (L.) Moench] is an important cereal crop for dryland areas in the United States and for small-holder farmers in Africa. Natural variation of sorghum grain composition (protein, fat, and starch) between accessions can be used for crop improvement, but the genetic controls are...

  2. De novo transcriptome assembly of Sorghum bicolor variety Taejin

    Directory of Open Access Journals (Sweden)

    Yeonhwa Jo

    2016-06-01

    Full Text Available Sorghum (Sorghum bicolor, also known as great millet, is one of the most popular cultivated grass species in the world. Sorghum is frequently consumed as food for humans and animals as well as used for ethanol production. In this study, we conducted de novo transcriptome assembly for sorghum variety Taejin by next-generation sequencing, obtaining 8.748 GB of raw data. The raw data in this study can be available in NCBI SRA database with accession number of SRX1715644. Using the Trinity program, we identified 222,161 transcripts from sorghum variety Taejin. We further predicted coding regions within the assembled transcripts by the TransDecoder program, resulting in a total of 148,531 proteins. We carried out BLASTP against the Swiss-Prot protein sequence database to annotate the functions of the identified proteins. To our knowledge, this is the first transcriptome data for a sorghum variety derived from Korea, and it can be usefully applied to the generation of genetic markers.

  3. Molecular cloning and expression of the calmodulin gene from guinea pig hearts.

    Science.gov (United States)

    Feng, Rui; Liu, Yan; Sun, Xuefei; Wang, Yan; Hu, Huiyuan; Guo, Feng; Zhao, Jinsheng; Hao, Liying

    2015-06-01

    The aim of the present study was to isolate and characterize a complementary DNA (cDNA) clone encoding the calmodulin (CaM; GenBank accession no. FJ012165) gene from guinea pig hearts. The CaM gene was amplified from cDNA collected from guinea pig hearts and inserted into a pGEM®-T Easy vector. Subsequently, CaM nucleotide and protein sequence similarity analysis was conducted between guinea pigs and other species. In addition, reverse transcription-polymerase chain reaction (RT-PCR) was performed to investigate the CaM 3 expression patterns in different guinea pig tissues. Sequence analysis revealed that the CaM gene isolated from the guinea pig heart had ∼90% sequence identity with the CaM 3 genes in humans, mice and rats. Furthermore, the deduced peptide sequences of CaM 3 in the guinea pig showed 100% homology to the CaM proteins from other species. In addition, the RT-PCR results indicated that CaM 3 was widely and differentially expressed in guinea pigs. In conclusion, the current study provided valuable information with regard to the cloning and expression of CaM 3 in guinea pig hearts. These findings may be helpful for understanding the function of CaM3 and the possible role of CaM3 in cardiovascular diseases.

  4. CLONING, EXPRESSION, AND MUTATIONAL ANALYSIS OF RAT S-ADENOSYL-1-METHIONINE: ARSENIC (III) METHYLTRANSFERASE

    Science.gov (United States)

    CLONING, EXPRESSION, AND MUTATIONAL ANALYSIS OF RAT S-ADENOSYL-L-METHIONINE: ARSENIC(III) METHYLTRANSFERASEStephen B. Waters, Ph.D., Miroslav Styblo, Ph.D., Melinda A. Beck, Ph.D., University of North Carolina at Chapel Hill; David J. Thomas, Ph.D., U.S. Environmental...

  5. Biological and water-use efficiencies of sorghum-groundnut intercrop

    African Journals Online (AJOL)

    In order to compare water-use efficiency of sole crops and intercrops, 2 experiments were conducted in 2 consecutive years with sorghum (Sorghum bicolor L. Moench) and groundnut (Arachis hypogaea L.) on a loamy, Grossarenic Paleudult. In a randomized block, split-plot design, sorghum (SS), groundnut (GG), ...

  6. Cloning and expression of cDNA coding for bouganin.

    Science.gov (United States)

    den Hartog, Marcel T; Lubelli, Chiara; Boon, Louis; Heerkens, Sijmie; Ortiz Buijsse, Antonio P; de Boer, Mark; Stirpe, Fiorenzo

    2002-03-01

    Bouganin is a ribosome-inactivating protein that recently was isolated from Bougainvillea spectabilis Willd. In this work, the cloning and expression of the cDNA encoding for bouganin is described. From the cDNA, the amino-acid sequence was deduced, which correlated with the primary sequence data obtained by amino-acid sequencing on the native protein. Bouganin is synthesized as a pro-peptide consisting of 305 amino acids, the first 26 of which act as a leader signal while the 29 C-terminal amino acids are cleaved during processing of the molecule. The mature protein consists of 250 amino acids. Using the cDNA sequence encoding the mature protein of 250 amino acids, a recombinant protein was expressed, purified and characterized. The recombinant molecule had similar activity in a cell-free protein synthesis assay and had comparable toxicity on living cells as compared to the isolated native bouganin.

  7. Pengujian Parameter Biji Sorghum dan Pengaruh Analisa Total Asam Laktat dan pH pada Tepung Sorghum Terfermentasi Menggunakan Baker’s Yeast (Saccharomyces Cereviceae

    Directory of Open Access Journals (Sweden)

    Amelinda Angelina

    2013-09-01

    Full Text Available Sorghum, Sorghum bicolor (L Moench, adalah sereal paling penting kelima setelah beras, jagung, barley dan gandum. Hasil penelitian menunjukkan bahwa kemampuan substitusi biji sorghum terhadap tepung terigu bisa mencapai 50-75%, walaupun nilai protein pembentuk glutennya tidak dapat menyamai tepung terigu. Tujuan dari penelitian ini adalah mempelajari pengaruh waktu fermentasi terhadap penurunan total asam laktat, nilai pH, dan jumlah total khamir (baker’s yeast tanpa menggunakan nutrient kimia tambahan . Analisa komposisi biji sorghum yang diinvestigasi dalam keadaan wet basis dari laboratorium menghasilkan kadar air, lemak, serat, protein, karbohidrat, dan abu masing-masing sebesar 12.85%, 3.10%, 0.56%, 5.87%, 75.82%, dan 1.79%. Untuk nilai energi total dengan metode bomb kalori didapatkan 4375.94 kcal/kg. Pengujian biji sorghum menghasilkan C-organik sebesar 12,47%. Berdasarkan analisa didapatkan hasil optimal dalam membuat tepung sorghum terfermentasi pada proses fermentasi 60 jam dengan jumlah yeast yang dihasilkan 1,7 x 105 sel/ml dengan kondisi yield % asam laktat 0,214%.

  8. Cloning, functional expression, and characterization of a PKA-activated gastric Cl- channel.

    Science.gov (United States)

    Malinowska, D H; Kupert, E Y; Bahinski, A; Sherry, A M; Cuppoletti, J

    1995-01-01

    cDNA encoding a Cl- channel was isolated from a rabbit gastric library, sequenced, and expressed in Xenopus oocytes. The predicted protein (898 amino acids, relative molecular mass 98,433 Da) was overall 93% similar to the rat brain ClC-2 Cl- channel. However, a 151-amino acid stretch toward the COOH-terminus was 74% similar to ClC-2 with six amino acids deleted. Two new potential protein kinase A (PKA) phosphorylation sites (also protein kinase C phosphorylation sites) were introduced. cRNA-injected Xenopus oocytes expressed a Cl- channel that was active at pHtrans 3 and had a linear current-voltage (I-V) curve and a slope conductance of 29 +/- 1 pS at 800 mM CsCl. A fivefold Cl- gradient caused a rightward shift in the I-V curve with a reversal potential of +30 +/- 3 mV, indicating anion selectivity. The selectivity was I- > Cl- > NO3-. The native and recombinant Cl- channel were both activated in vitro by PKA catalytic subunit and ATP. The electrophysiological and regulatory properties of the cloned and the native channel were similar. The cloned protein may be the Cl- channel involved in gastric HCl secretion.

  9. Optimization of cell line development in the GS-CHO expression system using a high-throughput, single cell-based clone selection system.

    Science.gov (United States)

    Nakamura, Tsuyoshi; Omasa, Takeshi

    2015-09-01

    Therapeutic antibodies are commonly produced by high-expressing, clonal and recombinant Chinese hamster ovary (CHO) cell lines. Currently, CHO cells dominate as a commercial production host because of their ease of use, established regulatory track record, and safety profile. CHO-K1SV is a suspension, protein-free-adapted CHO-K1-derived cell line employing the glutamine synthetase (GS) gene expression system (GS-CHO expression system). The selection of high-producing mammalian cell lines is a crucial step in process development for the production of therapeutic antibodies. In general, cloning by the limiting dilution method is used to isolate high-producing monoclonal CHO cells. However, the limiting dilution method is time consuming and has a low probability of monoclonality. To minimize the duration and increase the probability of obtaining high-producing clones with high monoclonality, an automated single cell-based clone selector, the ClonePix FL system, is available. In this study, we applied the high-throughput ClonePix FL system for cell line development using CHO-K1SV cells and investigated efficient conditions for single cell-based clone selection. CHO-K1SV cell growth at the pre-picking stage was improved by optimizing the formulation of semi-solid medium. The efficiency of picking and cell growth at the post-picking stage was improved by optimization of the plating time without decreasing the diversity of clones. The conditions for selection, including the medium formulation, were the most important factors for the single cell-based clone selection system to construct a high-producing CHO cell line. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  10. Optimization of extraction of polyphenols from Sorghum Moench ...

    African Journals Online (AJOL)

    phenolic acid were assayed using high performance liquid (HPLC). ... quantification of antioxidants and phenolic compounds from Sorghum M, ... Keywords: Response surface methodology, Sorghum moench, Polyphenols, Antioxidants.

  11. Clonal analysis of the T-cell response to in vivo expressed Mycobacterium tuberculosis protein Rv2034, using a CD154 expression based T-cell cloning method.

    Directory of Open Access Journals (Sweden)

    Susanna Commandeur

    Full Text Available Tuberculosis (TB, caused by Mycobacterium tuberculosis (Mtb, remains a leading cause of death worldwide. A better understanding of the role of CD4+ and CD8+ T cells, which are both important to TB protection, is essential to unravel the mechanisms of protection and to identify the key antigens seen by these T cells. We have recently identified a set of in vivo expressed Mtb genes (IVE-TB which is expressed during in vivo pulmonary infection in mice, and shown that their encoded antigens are potently recognized by polyclonal T cells from tuberculin skin test-positive, in vitro ESAT-6/CFP10-responsive individuals. Here we have cloned T cells specific for one of these newly identified in vivo expressed Mtb (IVE-TB antigens, Rv2034. T cells were enriched based on the expression of CD154 (CD40L, which represents a new method for selecting antigen-specific (low frequency T cells independent of their specific function. An Rv2034-specific CD4+ T-cell clone expressed the Th1 markers T-bet, IFN-γ, TNF-α, IL-2 and the cytotoxicity related markers granzyme B and CD107a as measured by flow cytometry. The clone specifically recognized Rv2034 protein, Rv2034 peptide p81-100 and Mtb lysate. Remarkably, while the recognition of the dominant p81-100 epitope was HLA-DR restricted, the T-cell clone also recognized a neighboring epitope (p88-107 in an HLA-DR- as well as HLA-DQ1-restricted fashion. Importantly, the T-cell clone was able to inhibit Mtb outgrowth from infected monocytes significantly. The characterization of the polyfunctional and Mtb inhibitory T-cell response to IVE-TB Rv2034 at the clonal level provides detailed further insights into the potential of IVE-TB antigens as new vaccine candidate antigens in TB. Our new approach allowed the identification of T-cell subsets that likely play a significant role in controlling Mtb infection, and can be applied to the analysis of T-cell responses in patient populations.

  12. Peculiarities in covering the requirements for seed material of sorghum crops

    Directory of Open Access Journals (Sweden)

    С. І. Мельник

    2017-12-01

    Full Text Available Purpose. To assess the demand for sorghum seed material and sufficiency of domestic seeds. Results. The analysis of the State register for the period of 2002–2012 showed that there was the tendency not only towards increasing quantity of sorghum crops in general but their substitution by hybrids of foreign breeding. During the period from 2002 to 2017, 72 sorghum varieties were entered on the State register in total, among them only 12 varieties were of domestic breeding, the rest 60 was presented by foreign breeding institutions. Investigation results allowed to determine that the production of base and prebase seeds of sorghum in 2010 amounted to 1,3 t, in 2016 was 43 t. During the same period the production of sugar sorghum increased from 0,2 to 12,0 t, grass sorghum – from 4,0 to 83 t. In 2017, requirements of acreage of such crops as grass sorghum and broomcorn were completely satisfied by the amount of grown seeds. At the same time, the need for seeds of sorghum and sugar sorghum can not be covered completely at the expense of domestic varieties reproduction. In 2017, general demand for sorghum seeds was 400,5 t, among which only 42,0 t was of domestic production. The rest demand for seeds will be met at the expense of import of foreign breeding seeds into the country to be grown and prepared for sowing abroad. Conclusions. In the Register of plant varieties suitable for dissemination in Ukraine, there are 72 sorghum varieties among them only 12 varieties were of domestic breeding, that is 17%, as compared to 83% of recommended great sorghum varieties of foreign breeding. In Ukraine, the area occupied by sorghum cultivation was 22,8 thou ha in 2005, up to 2017 it increased to 89,0 thou ha, and accordingly the demand for seeds run up from 102,6 to 400,5 t. The area occupied by the sugar sorghum in 2005 amounted to only 2,6 thou ha, in 2017 – 20,0 thou ha, that accordingly determined increase of demand for seed material from 13,0 to 99

  13. Cloning, bacterial expression and crystallization of Fv antibody fragments

    Science.gov (United States)

    E´, Jean-Luc; Boulot, Ginette; Chitarra, V´ronique; Riottot, Marie-Madeleine; Souchon, H´le`ne; Houdusse, Anne; Bentley, Graham A.; Narayana Bhat, T.; Spinelli, Silvia; Poljak, Roberto J.

    1992-08-01

    The variable Fv fragments of antibodies, cloned in recombinant plasmids, can be expressed in bacteria as functional proteins having immunochemical properties which are very similar or identical with those of the corresponding parts of the parent eukaryotic antibodies. They offer new possibilities for the study of antibody-antigen interactions since the crystals of Fv fragments and of their complexes with antigen reported here diffract X-rays to a higher resolution that those obtained with the cognate Fab fragments. The Fv approach should facilitate the structural study of the combining site of antibodies and the further characterization of antigen-antibody interactions by site-directed mutagenesis experiments.

  14. Cloning, expression, and crystallization of Cpn60 proteins from Thermococcus litoralis.

    Science.gov (United States)

    Osipiuk, J; Sriram, M; Mai, X; Adams, M W; Joachimiak, A

    2000-01-01

    Two genes of the extreme thermophilic archaeon Thermococcus litoralis homologous to those that code for Cpn60 chaperonins were cloned and expressed in Escherichia coli. Each of the Cpn60 subunits as well as the entire Cpn60 complex crystallize in a variety of morphological forms. The best crystals diffract to 3.6 A resolution at room temperature and belong to the space group 1422 with unit cell parameters a = b = 193.5 A, c = 204.2 A.

  15. Enhanced ethanol production from stalk juice of sweet sorghum by ...

    African Journals Online (AJOL)

    user

    2012-03-15

    Mar 15, 2012 ... Sweet sorghum (sugar sorghum, Sorghum bicolor) is one kind of non-grain energy ... government that only ''non-grain” materials can be used ... In this work, ... inoculated (10%, v/v) into fermentation medium prepared with the.

  16. Cloning, expression, purification and crystallization of dihydrodipicolinate synthase from the grapevine Vitis vinifera

    International Nuclear Information System (INIS)

    Atkinson, Sarah C.; Dogovski, Con; Newman, Janet; Dobson, Renwick C. J.; Perugini, Matthew A.

    2011-01-01

    Dihydrodipicolinate synthase from the common grapevine V. vinifera has been cloned, expressed, purified and crystallized in the presence of the substrate pyruvate by in-drop hexahistidine-tag cleavage. A diffraction data set has been collected to a resolution of 2.2 Å. Dihydrodipicolinate synthase (DHDPS) catalyses the first committed step of the lysine-biosynthesis pathway in bacteria, plants and some fungi. This study describes the cloning, expression, purification and crystallization of DHDPS from the grapevine Vitis vinifera (Vv-DHDPS). Following in-drop cleavage of the hexahistidine tag, cocrystals of Vv-DHDPS with the substrate pyruvate were grown in 0.1 M Bis-Tris propane pH 8.2, 0.2 M sodium bromide, 20%(w/v) PEG 3350. X-ray diffraction data in space group P1 at a resolution of 2.2 Å are presented. Preliminary diffraction data analysis indicated the presence of eight molecules per asymmetric unit (V M = 2.55 Å 3 Da −1 , 52% solvent content). The pending crystal structure of Vv-DHDPS will provide insight into the molecular evolution in quaternary structure of DHDPS enzymes

  17. Effect of Fungicide Applications on Grain Sorghum (Sorghum bicolor L. Growth and Yield

    Directory of Open Access Journals (Sweden)

    Dan D. Fromme

    2017-01-01

    Full Text Available Field studies were conducted in the upper Texas Gulf Coast and in central Louisiana during the 2013 through 2015 growing seasons to evaluate the effects of fungicides on grain sorghum growth and development when disease pressure was low or nonexistent. Azoxystrobin and flutriafol at 1.0 L/ha and pyraclostrobin at 0.78 L/ha were applied to the plants of two grain sorghum hybrids (DKS 54-00, DKS 53-67 at 25% bloom and compared with the nontreated check for leaf chlorophyll content, leaf temperature, and plant lodging during the growing season as well as grain mold, test weight, yield, and nitrogen and protein content of the harvested grain. The application of a fungicide had no effect on any of the variables tested with grain sorghum hybrid responses noted. DKS 53-67 produced higher yield, greater test weight, higher percent protein, and N than DKS 54-00. Results of this study indicate that the application of a fungicide when little or no disease is present does not promote overall plant health or increase yield.

  18. Cloning, Expression, Characterization, and Computational Approach for Cross-Reactivity Prediction of Manganese Superoxide Dismutase Allergen from Pistachio Nut

    Directory of Open Access Journals (Sweden)

    Reihaneh Noorbakhsh

    Full Text Available ABSTRACT: Background: Tree nut allergy is one of the common potentially life-threatening food allergies in children and adults. Recombinant food allergens offer new perspectives to solve problems of clinical and molecular allergology in diagnosis, research, and therapy of food allergies. So far, superoxide dismutase (s has been identified as a panallergen and studied in different allergenic sources. Manganese Superoxide Dismutase (MnSOD has also been reported in pistachio that may cause allergic reactions in atopic subjects. The aim of this study was to describe the cloning, expression, and purification of MnSOD from pistachio nut. Methods: The pistachio MnSOD was cloned and expressed in E. coli BL21 (DE3 using a vector pET-32b (+. A recombinant protein was purified by metal precipitation. The protein immunoreactivity was evaluated using patients' IgE binding by means of ELISA and immunoblotting assays. Results: The MnSOD gene from pistachio was successfully cloned and expressed in E. coli. The purified pistachio MnSOD was recognized by IgE in 10 (40% out of the 25 sera tested. Our results also showed that this protein might trigger some cross-reactions toward IgE antibodies and thus could be considered as a panallergen. Conclusions: For the first time recombinant manganese superoxide dismutase from nut source was expressed as a possible allergen. This pistachio allergen could be a possible basis for cross-reactivity with MnSOD from other sources. KEY WORDS: cloning, cross-reaction, Manganese Superoxide Dismutase (MnSOD, pistachio (Pistacia vera, recombinant allergen

  19. Next-generation sequencing technology for genetics and genomics of sorghum

    DEFF Research Database (Denmark)

    Luo, Hong; Mocoeur, Anne Raymonde Joelle; Jing, Hai-Chun

    2014-01-01

    and grain sorghum. NGS has also been used to examine the transcriptomes of sorghum under various stress conditions. Besides identifying interesting transcriptonal adpatation to stress conditions, these study show that sugar could potentially act as an osmitic adjusting factor via transcriptional regulation....... Furthermore, miRNAs are found to be important adaptation to both biotic and abiotic stresses in sorghum. We discuss the use of NGS for further genetic improvement and breeding in sorghum....

  20. SALL4 expression in gonocytes and spermatogonial clones of postnatal mouse testes.

    Directory of Open Access Journals (Sweden)

    Kathrin Gassei

    Full Text Available The spermatogenic lineage is established after birth when gonocytes migrate to the basement membrane of seminiferous tubules and give rise to spermatogonial stem cells (SSC. In adults, SSCs reside within the population of undifferentiated spermatogonia (A(undiff that expands clonally from single cells (A(single to form pairs (A(paired and chains of 4, 8 and 16 A(aligned spermatogonia. Although stem cell activity is thought to reside in the population of A(single spermatogonia, new research suggests that clone size alone does not define the stem cell pool. The mechanisms that regulate self-renewal and differentiation fate decisions are poorly understood due to limited availability of experimental tools that distinguish the products of those fate decisions. The pluripotency factor SALL4 (sal-like protein 4 is implicated in stem cell maintenance and patterning in many organs during embryonic development, but expression becomes restricted to the gonads after birth. We analyzed the expression of SALL4 in the mouse testis during the first weeks after birth and in adult seminiferous tubules. In newborn mice, the isoform SALL4B is expressed in quiescent gonocytes at postnatal day 0 (PND0 and SALL4A is upregulated at PND7 when gonocytes have colonized the basement membrane and given rise to spermatogonia. During steady-state spermatogenesis in adult testes, SALL4 expression overlapped substantially with PLZF and LIN28 in A(single, A(paired and A(aligned spermatogonia and therefore appears to be a marker of undifferentiated spermatogonia in mice. In contrast, co-expression of SALL4 with GFRα1 and cKIT identified distinct subpopulations of A(undiff in all clone sizes that might provide clues about SSC regulation. Collectively, these results indicate that 1 SALL4 isoforms are differentially expressed at the initiation of spermatogenesis, 2 SALL4 is expressed in undifferentiated spermatogonia in adult testes and 3 SALL4 co-staining with GFRα1 and c

  1. Nutrient and carbohydrate partitioning in sorghum stover

    International Nuclear Information System (INIS)

    Powell, J.M.; Hons, F.M.; McBee, G.G.

    1991-01-01

    Sorghum [Sorghum bicolor (L.) Moench] stover has been demonstrated to be a potential biomass energy source. Complete aboveground crop removal, however, can result in soil degradation. Differential dry matter, nutrient, and carbohydrate partitioning by sorghum cultivars may allow management strategies that return certain parts to the field while removing other portions for alternative uses, such as energy production. A field study was conducted to determine N,P,K, nonstructural carbohydrate, cellulose hemicellulose, and lignin distributions in stover of three diverse sorghum cultivars of differing harvest indices. Determinations were based on total vegetative biomass; total blades; total stalks; and upper middle, and lower blades and stalks. Concentrations of N and P were higher in blades than stalks and generally declines from upper to lower stover parts. Large carbohydrate and lignin concentration differences were observed on the basis of cultivar and stover part. Greater nutrient partitioning to the upper third of the intermediate and forage-type sorghum stovers was observed as compared to the conventional grain cultivar. Stover carbohydrates for all cultivars were mainly contained in the lower two-thirds of the stalk fraction. A system was proposed for returning upper stover portion to soil, while removing remaining portions for alternative uses

  2. Tapping the US sweet sorghum collection to identify biofuel germplasm

    Science.gov (United States)

    The narrow genetic base in sweet sorghum [Sorghum bicolor (L.) Moench] breeding programs is limiting the development of new varieties for biofuel production. Therefore, the identification of genetically diverse sweet sorghum germplasm in the U.S. National Plant Germplasm System (NPGS) collection is...

  3. High-throughput cloning and expression in recalcitrant bacteria

    NARCIS (Netherlands)

    Geertsma, Eric R.; Poolman, Bert

    We developed a generic method for high-throughput cloning in bacteria that are less amenable to conventional DNA manipulations. The method involves ligation-independent cloning in an intermediary Escherichia coli vector, which is rapidly converted via vector-backbone exchange (VBEx) into an

  4. An investigative study of indigenous sweet sorghum varieties for bioethanol production: the case of Kenya local sorghum varieties

    Energy Technology Data Exchange (ETDEWEB)

    Wangai, L.K.; Mbeo, C.O. [Kenya Industrial Research and Development Inst., Nairobi (Kenya); Kamau, C.K. [Kenya Agricurtural Research Inst.(s), Machakos (Kenya)

    2012-11-01

    There are over 500 sorghum genotypes grown locally in Kenya. This study was an investigation and selection of suitable sorghum genotypes for sustainable bio-ethanol production in Kenya. For the study, 500 genotypes of sorghum were planted and grown using the recommended agricultural practices. Random sampling of 230 genotypes was done and the samples analysed for juice and sugar content. The 26 best yielding genotypes were selected and grown again in duplicate for further detailed study. Data on date of flowering, pest resistance, {sup 0}brix, wet and dry weight, plant population, ratooning, grain yield and juice yield and juice sugar content were recorded and analyzed using GENstat. Sampling was done for each genotype when about 50% of the crop had flowered and there after, every 2 weeks until the grains dried. Crushing was done with a three roller mill crusher [8]. The sugar content was measured using a digital refractometer. Sugar yield obtained ranged between 10.3{sup 0}Brix and 19.3{sup 0}Brix and juice yield between 268 litres/hectare and 11390 litres/hectare. Five indigenous sorghum varieties, GBK-007130, GBK-007076, GBK-007102, GBK-007296, GBK-007098 were found to have the highest sugar and juice yields and were considered the most suitable sweet sorghum genotypes among those studied, for bio-ethanol production in Kenya.

  5. Identification, cloning, and expression of a GHF9 cellulase from Tribolium castaneum (Coleoptera: Tenebrionidae)

    Science.gov (United States)

    The availability of sequenced insect genomes has allowed for discovery and functional characterization of novel genes and proteins. We report use of the Tribolium castaneum (Herbst) (red flour beetle) genome to identify, clone, express, and characterize a novel endo-ß-1,4-glucanase we named TcEG1 (...

  6. In Vitro Screening for Drought Tolerance in Different Sorghum (Sorghum bicolor (L. Moench Varieties

    Directory of Open Access Journals (Sweden)

    Yohannes Tsago

    2013-08-01

    Full Text Available Drought is one of the complex environmental factors affecting growth and yield of sorghum in arid and semi-arid areas of the world. Sixteen elite sorghum (Sorghum bicolor (L Moench genotypes were evaluated for their genetic potential to drought tolerance at callus induction and plant regeneration stage for drought tolerance. The non-ionic water soluble polymer polyethylene glycol (PEG of molecular weight 6000 was used as osmoticum to simulate water stress. The factorial experiment was laid down in a completely randomized design which comprised of a combination of two factors (genotypes and five PEG stress level; 0, 0.5, 1.0, 1.5, and 2.0% (w/v treatments. Data were recorded for callus induction efficiency, callus fresh weight, embryogenic callus percentage and plant regeneration percentage. Significant differences were observed among the genotypes, treatments and their interactions for the evaluated plant traits suggesting a great amount of variability for drought tolerance in sorghum. The correlation analysis also revealed strong and significant association between embryogenic callus percent and plant regeneration percent as well as between embryogenic callus percent and plant regeneration percent. By taking into consideration all the measured traits, Mann Whitney rank sum test revealed that 76T1#23 and Teshale followed by Meko, Gambella-1107 and Melkam showed better drought stress tolerance. Therefore they are recommended to be used as parents for genetic analysis, gene mapping and improvement of drought tolerance while Chelenko, Hormat and Raya appear to be drought sensitive.

  7. Cloning and Molecular Analyses of a Gibberellin 20-Oxidase Gene Expressed Specifically in Developing Seeds of Watermelon1

    Science.gov (United States)

    Kang, Hong-Gyu; Jun, Sung-Hoon; Kim, Junyul; Kawaide, Hiroshi; Kamiya, Yuji; An, Gynheung

    1999-01-01

    To understand the biosynthesis and functional role of gibberellins (GAs) in developing seeds, we isolated Cv20ox, a cDNA clone from watermelon (Citrullus lanatus) that shows significant amino acid homology with GA 20-oxidases. The complementary DNA clone was expressed in Escherichia coli as a fusion protein, which oxidized GA12 at C-20 to the C19 compound GA9, a precursor of bioactive GAs. RNA-blot analysis showed that the Cv20ox gene was expressed specifically in developing seeds. The gene was strongly expressed in the integument tissues, and it was also expressed weakly in inner seed tissues. In parthenocarpic fruits induced by 1-(2-chloro-4-pyridyl)-3-phenylurea treatment, the expression pattern of Cv20ox did not change, indicating that the GA 20-oxidase gene is expressed primarily in the maternal cells of developing seeds. The promoter of Cv20ox was isolated and fused to the β-glucuronidase (GUS) gene. In a transient expression system, β-glucuronidase staining was detectable only in the integument tissues of developing watermelon seeds. PMID:10517828

  8. Structure and chemistry of the sorghum grain

    Science.gov (United States)

    Sorghum is grown around the world and often under harsh and variable environmental conditions. Combined with the high degree of genetic diversity present in sorghum, this can result in substantial variability in grain composition and grain quality. While similar to other cereal grains such as maize ...

  9. Expression of brown-midrib in a spontaneous sorghum mutant is linked to a 5'-UTR deletion in lignin biosynthesis gene SbCAD2

    Science.gov (United States)

    Brown midrib (bmr) mutants in sorghum (Sorghum bicolor (L.) Moench) and several other C4 grasses are associated with reduced lignin concentration, altered lignin composition and improved cell wall digestibility, which are desirable properties in biomass development for the emerging lignocellulosic b...

  10. Cloning, high-level expression, purification and crystallization of peptide deformylase from Leptospira interrogans.

    Science.gov (United States)

    Li, Yikun; Ren, Shuangxi; Gong, Weimin

    2002-05-01

    A new peptide deformylase (PDF; EC 3.5.1.27) gene from Leptospira interrogans was identified and cloned into expression plasmid pET22b(+) and was highly expressed in Escherichia coli BL21(DE3). With DEAE-Sepharose anion-exchange chromatography followed by Superdex G-75 size-exclusion chromatography, 60 mg of PDF from L. interrogans was purified from 1 l of cell culture. Crystallization screening of the purified enzyme resulted in two crystal forms, from one of which a 3 A resolution X-ray diffraction data set has been collected.

  11. The regulation of science and the Charter of Rights: would a ban on non-reproductive human cloning unjustifiably violate freedom of expression?

    Science.gov (United States)

    Billingsley, Barbara; Caulfield, Timothy

    2004-01-01

    Non-Reproductive Human Cloning (NRHC) allows researchers to develop and clone cells, including non-reproductive cells, and to research the etiology and transmission of disease. The ability to clone specific stem cells may also allow researchers to clone cells with genetic defects and analyze those cells with more precisions. Despite those potential benefits, Parliament has banned such cloning due to a myriad of social and ethical concerns. In May 2002, the Canadian Government introduced Bill C-13 on assisted human reproductive technologies. Bill C-13 deals with both the scientific and the clinical use of human reproductive materials, and it prohibits a number of other activities, including NRHC. Although the Supreme Court of Canada has never ruled on whether scientific experiments area form of expression, academic support exists for this notion. The authors go through the legal analysis that would be required to find that scientific experiments are expression, focusing in part on whether NRHC could be considered violent and thus fall outside the protection of section 2(b). The latter question is complicated by the ongoing policy debate over whether an "embryonic cell" is property of human life. The authors then consider whether a ban on NRHC could be justified under section 1 of the Charter. They conclude that both the breadth of the legislative purpose and the proportionality of the measure are problematic. Proportionality is a specific concern because the ban could be viewed as an outright denial of scientific freedom of expression. Although consistent with current jurisprudence on freedom of expression, this paper runs against the flow of government policy in the areas of regulation and prohibition of non-reproductive human cloning. As there has been no Charter litigation to date on whether scientific research is a form of expression, the authors introduce a new way of looking at the legality of the regulation of new reproductive technologies.

  12. EVALUATION OF TWO VARIETIES OF SORGHUM FOR STARCH EXTRACTION

    Directory of Open Access Journals (Sweden)

    Leyanis Rodríguez Rodríguez

    2015-01-01

    Full Text Available In Cuba, the wet milling process for the extraction of starch is made from corn, cereal which is currently imported, that is why it is required to substitute it for another grain of national production as it is the case of sorghum. Given the similarities of the two grains in their starch content and considering the potential of sorghum for the food industry, it is developed in this work an assessment process, taking into account two sorghum varieties: red (CIAPR-132 and white (UDG-110. In this sense, a factorial design of the type 2k-1 is made, where the independent variables of most influence in the laboratory process are considered, such as: (x1 type of sorghum, (x2 soaking time and (x3 solution concentration. It is considered that there is no interaction between them and it is taken as the response variable the starch yield in the extraction process. We conclude that the type of sorghum and soaking time are the most influential variables, obtaining the best results for white sorghum subjected for 48 hours to soak in a solution of SO2 at a concentration of 1800 ppm.

  13. Performance of Sorghum Varieties under Variable Rainfall in Central Tanzania.

    Science.gov (United States)

    Msongaleli, Barnabas M; Tumbo, S D; Kihupi, N I; Rwehumbiza, Filbert B

    2017-01-01

    Rainfall variability has a significant impact on crop production with manifestations in frequent crop failure in semiarid areas. This study used the parameterized APSIM crop model to investigate how rainfall variability may affect yields of improved sorghum varieties based on long-term historical rainfall and projected climate. Analyses of historical rainfall indicate a mix of nonsignificant and significant trends on the onset, cessation, and length of the growing season. The study confirmed that rainfall variability indeed affects yields of improved sorghum varieties. Further analyses of simulated sorghum yields based on seasonal rainfall distribution indicate the concurrence of lower grain yields with the 10-day dry spells during the cropping season. Simulation results for future sorghum response, however, show that impacts of rainfall variability on sorghum will be overridden by temperature increase. We conclude that, in the event where harms imposed by moisture stress in the study area are not abated, even improved sorghum varieties are likely to perform poorly.

  14. Isolation and partial characterization of peripheral blood CD4+ T cell clones expressing γδT cell receptors

    International Nuclear Information System (INIS)

    Kyoizumi, Seishi; Akiyama, Mitoshi; Hirai, Yuko; Kusunoki, Yoichiro.

    1990-06-01

    Rare T cell clones bearing both CD4 and T cell receptors (TCRγ and TCRδ) were obtained from human peripheral blood by cell sorting using anti-CD4 and anti-TCRδ1 antibodies. All the clones established were reactive with anti-TCRγδ1 antibody, whereas only about 20 % of the clones showed reactivity with anti-δTCS1 antibody. Unlike CD4 + T cells bearing TCRαβ, all the clones tested were lectin-dependent and showed CD3 antibody-redirected cytolytic activity. About 60 % exhibited natural killer cell-like activity. Immunoprecipitation analysis of TCRγδ showed that each clone expressed either a disulfide-linked or nondisulfide-linked heterodimer consisting of 37-44 kilodalton TCRγ and TCRδ chains. Southern blot analyses of TCRγ and TCRδ genes revealed some identical rearrangement patterns, suggesting the limited heterogeneity of CD4 + TCRγδ + T cells in peripheral blood. (author)

  15. Evaluation of the multi-seeded (msd) mutant of sorghum for ethanol production

    Science.gov (United States)

    Grain sorghum [Sorghum bicolor (L.) Moench], a cost effective crop in semiarid regions, is an underestimated supplement to corn in starch based ethanol production. Twenty three multi-seeded (msd) mutant sorghums and one wild type sorghum BTx623 were evaluated for ethanol production and effect of che...

  16. Molecular cloning, expression analysis and sequence prediction of ...

    African Journals Online (AJOL)

    CCAAT/enhancer-binding protein beta as an essential transcriptional factor, regulates the differentiation of adipocytes and the deposition of fat. Herein, we cloned the whole open reading frame (ORF) of bovine C/EBPβ gene and analyzed its putative protein structures via DNA cloning and sequence analysis. Then, the ...

  17. Comparative analysis of CDPK family in maize, Arabidopsis, rice and sorghum revealed potential targets for drought tolerance improvement

    Science.gov (United States)

    Mittal, Shikha; Mallikarjuna, Mallana Gowdra; Rao, Atmakuri R.; Jain, Prashant A.; Dash, Prasanta K.; Thirunavukkarasu, Nepolean

    2017-12-01

    Calcium dependent protein kinases (CDPKs) play major role in regulation of plant growth and development in response to various stresses including drought. A set of 32 CDPK genes identified in maize were further used for searching of orthologs in the model plant Arabidopsis (72) and major food crops such as rice (78) and sorghum (91). We comprehensively investigated the phylogenetic relationship, annotations, gene duplications, gene structure, divergence time, 3-D protein structures and tissue-specific drought induced expression of CDPK genes in all four species. Variation in intron frequency among these species likely contributed to the functional diversity of CDPK genes to various stress responses. Protein kinase and protein kinase C phosphorylation site domains were the most conserved motifs identified in all species. Four groups were identified from the sequence-based phylogenetic analysis, in which maize CDPKs were clustered in group III. The time of divergence (Ka/Ks) analysis revealed that the CDPKs were evolved through stabilizing selection. Expression data showed that the CDPK genes were highly expressed in leaf of maize, rice, and sorghum whereas in Arabidopsis the maximum expression was observed in root. 3-D protein structure were predicted for the nine genes (Arabidopsis: 2, maize: 2, rice: 3 and sorghum: 2) showing differential expression in at least three species. The predicted 3-D structures were further evaluated and validated by Ramachandran plot, ANOLEA, ProSA and Verify-3D. The superimposed 3-D structure of drought-related orthologous proteins retained similar folding pattern owing to their conserved nature. Functional annotation revealed the involvement of CDPK genes in various pathways such as osmotic homeostasis, cell protection and root growth. The interactions of CDPK genes in various pathways play crucial role in imparting drought tolerance through different ABA and MAPK signalling cascades. Our studies suggest that these selected candidate

  18. Sorghum bagasse as substrate for cellulase production by submerged and solid-state cultures of Trichoderma

    Directory of Open Access Journals (Sweden)

    Teodor Vintilă

    2014-05-01

    Full Text Available Sweet sorghum bagasse was used as cellulosic substrate in submerged and solid-state cultures of Trichoderma for cellulase production. Submerged liquid cultures (SLC were obtained by inoculation of Mandels media containing 1% cellulose with spores suspension of Trichoderma. Solid-state cultures (SSC were carried out in Erlenmayer flasks, where the substrate was distributed 1 cm layers. Comparing the yields of cellulases produced by Trichoderma strains in the systems applied in this study, using as substrate sorghum bagasse, we found the solid-state cultures as the system to produce the highest cellulase yields. The local strain of T. viride CMIT3.5. express high productivity in SSC system in laboratory conditions. The cellulolytic enzymes have maximum activity at 50oC, pH 4,8. The results recommend solid-state cultures of Trichoderma on sorghum bagasse as systems for producing cellulolytic products with higher activity than submerged cultures of Trichoderma on the same substrate.

  19. Cloning, expression, and characterization of cadmium and manganese uptake genes from Lactobacillus plantarum

    Energy Technology Data Exchange (ETDEWEB)

    Hao, Z.; Chen, S.; Wilson, D.B.

    1999-11-01

    An Mn{sup 2+} and Cd{sup 2+} uptake gene, mntA, was cloned from Lactobacillus plantarum ATCC 14917 into Escherichia coli. Its expression conferred on E. coli cells increased Cd{sup 2+} sensitivity as well as energy-dependent Cd{sup 2+} uptake activity. Both transcription and translation of mntA were induced by Mn{sup 2+} starvation in L. plantarum, as indicated by reverse transcriptase PCR and immunoblotting. Two Cd{sup 2+} uptake systems have been identified in L. plantarum: one is a high-affinity Mn{sup 2+} and Cd{sup 2+} uptake system that is expressed in Mn{sup 2+}-starved cells, and the other is a nonsaturable Cd{sup 2+} uptake system that is expressed in Cd{sup 2+}-sufficient cells. MntA was not detected in an Mn{sup 2+}-dependent mutant of L. plantarum which had lost high-affinity Mn{sup 2+} and Cd{sup 2+} uptake activity. The results suggest that mntA is the gene encoding the high-affinity Mn{sup 2+} and Cd{sup 2+} transporter. On the basis of its predicted amino acid sequence, MntA belongs to the family of P-type cation-translocating ATPases. The topology and potential Mn{sup 2+}- and Cd{sup 2+}-binding sites of MntA are discussed. A second clone containing a low-affinity Cd{sup 2+} transport system was also isolated.

  20. Molecular cloning of chicken metallothionein. Deduction of the complete amino acid sequence and analysis of expression using cloned cDNA

    Energy Technology Data Exchange (ETDEWEB)

    Wei, D; Andrews, G K

    1988-01-25

    A cDNA library was constructed using RNA isolated from the livers of chickens which had been treated with zinc. This library was screened with a RNA probe complementary to mouse metallothionein-I (MT), and eight chicken MT cDNA clones were obtained. All of the cDNA clones contained nucleotide sequences homologous to regions of the longest (375 bp) cDNA clone. The latter contained an open reading frame of 189 bp, and the deduced amino acid sequence indicates a protein of 63 amino acids of which 20 are cysteine residues. Amino acid composition and partial amino acid sequence analyses of purified chicken MT protein agreed with the amino acid composition and sequence deduced from the cloned cDNA. Amino acid sequence comparison establish that chicken MT shares extensive homology with mammalian MTs. Southern blot analysis of chicken DNA indicates that the chicken MT gene is not a part of a large family of related sequences, but rather is likely to be a unique gene sequence. In the chicken liver, levels of chicken MT mRNA were rapidly induced by metals (Cd/sup 2 +/, Zn/sup 2 +/, Cu/sup 2 +/), glucocorticoids and lipopolysaccharide. MT mRNA was present in low levels in embryonic liver and increased to high levels during the first week after hatching before decreasing again to the basal levels found in adult liver. The results of this study establish that MT is highly conserved between birds and mammals and is regulated in the chicken by agents which also regulate expression of mammalian MT genes. However, in contrast to the mammals, the results suggest the existence of a single isoform of MT in the chicken.

  1. Bioethanol production from dried sweet sorghum stalk

    Energy Technology Data Exchange (ETDEWEB)

    Almodares, A.; Etemadifar, Z.; Ghoreishi, F.; Yosefi, F. [Biology Dept. Univ. of Isfahan, Isfahan (Iran, Islamic Republic of)], e-mail: aalmodares@yahoo.com

    2012-11-01

    Bioethanol as a renewable transportation fuel has a great potential for energy and clean environment. Among crops sweet sorghum is one of the best feedstock for ethanol production under hot and dry climatic conditions. Because it has higher tolerance to salt and drought comparing to sugarcane and corn that are currently used for bio-fuel production in the world. Generally mills are used to extract the juice from sweet sorghum stalks. Three roller mills extract around nearly 50 percent of the juice and more mills is needed to extract higher percentage of the juice. More over under cold weather the stalks become dry and juice is not extracted from the stalk, therefore reduce harvesting period. In this study stalks were harvested, leaves were stripped from the stalks and the stalks were chopped to nearly 4 mm length and sun dried. The dry stalks were grounded to 60 mesh powder by a mill. Fermentation medium consists of 15-35% (w/w) sweet sorghum powder, micronutrients and active yeast inoculum from 0.5-1% (w/w) by submerge fermentation method. The fermentation time and temperature were 48-72 hours and 30 deg, respectively. The results showed the highest amount of ethanol (14.5 % w/w sorghum) was produced with 10% sweet sorghum powder and 1% of yeast inoculum, three day fermentation at 30 deg.

  2. Sorghum stem yield and soluble carbohydrates under different ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-11-19

    Nov 19, 2008 ... Key words: Sweet sorghum, grain sorghum, salinity, stem yield, ... The effect of salinity on the stem yield and sucrose was .... growth and polyamine metabolism in two citrus rootstocks with ... Growth and osmoregulation in two.

  3. Expression cloning of camelid nanobodies specific for Xenopus embryonic antigens.

    Directory of Open Access Journals (Sweden)

    Keiji Itoh

    Full Text Available Developmental biology relies heavily on the use of conventional antibodies, but their production and maintenance involves significant effort. Here we use an expression cloning approach to identify variable regions of llama single domain antibodies (known as nanobodies, which recognize specific embryonic antigens. A nanobody cDNA library was prepared from lymphocytes of a llama immunized with Xenopus embryo lysates. Pools of bacterially expressed cDNAs were sib-selected for the ability to produce specific staining patterns in gastrula embryos. Three different nanobodies were isolated: NbP1 and NbP3 stained yolk granules, while the reactivity of NbP7 was predominantly restricted to the cytoplasm and the cortex. The isolated nanobodies recognized specific protein bands in immunoblot analysis. A reverse proteomic approach identified NbP1 target antigen as EP45/Seryp, a serine protease inhibitor. Given the unique stability of nanobodies and the ease of their expression in diverse systems, we propose that nanobody cDNA libraries represent a promising resource for molecular markers for developmental biology.

  4. Apoptosis Gene Hunting Using Retroviral Expression Cloning: Identification of Vacuolar ATPase Subunit E

    Directory of Open Access Journals (Sweden)

    Claire L. Anderson

    2003-01-01

    Full Text Available Over the past 10-15 years there has been an explosion of interest in apoptosis. The delayed realisation that cell death is an essential part of life for any multicellular organism has meant that, despite the recent and rapid developments of the last decade, the precise biochemical pathways involved in apoptosis remain incomplete and potentially novel genes may, as yet, remain undiscovered. The hunt is therefore on to bridge the remaining gaps in our knowledge. Our contribution to this research effort utilises a functional cloning approach to isolate important regulatory genes involved in apoptosis. This mini-review focuses on the use and advantages of a retroviral expression cloning strategy and describes the isolation and identification of one such potential apoptosis regulatory gene, namely that encoding vacuolar ATPase subunit E.

  5. Purification, cDNA Cloning, and Developmental Expression of the Nodule-Specific Uricase from Phaseolus vulgaris L. 1

    Science.gov (United States)

    Sánchez, Federico; Campos, Francisco; Padilla, Jaime; Bonneville, Jean-Marc; Enríquez, Consuelo; Caput, Daniel

    1987-01-01

    Nodule-specific uricase (uricase II) from Phaseolus vulgaris L. was purified to homogeneity by chromatographic methods. Purification data indicated that uricase II is approximately 2% of the total soluble protein from mature nodules. Specific antiserum was raised and used to determine the developmental expression and for immunoselection of polysomes. Uricase II was antigenically detected early in nodule development, 2 to 3 days before nitrogen fixation. Uricase-encoding cDNA clones were isolated by hybridizing a nodule-specific pUC9 cDNA library with labeled mRNA from immunoselected polysomes and a 35,000 molecular weight uricase II-encoding cDNA from soybean. An homologous clone (pNF-UR07) was used to assess the expression pattern of the specific transcript during development. Northern-blot analysis indicated that uricase II mRNA is exclusively expressed in nodule tissue. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 PMID:16665575

  6. CLONING, EXPRESSION, AND CHARACTERIZATION OF RAT S-ADENOSYL-L-METHIONINE: ARSENIC (III) METHYLTRANSFERASE (CYT19)

    Science.gov (United States)

    CLONING, EXPRESSION, AND CHARACTERIZATION OF RAT S-ADENOSYL-L-METHIONINE: ARSENIC(III) METHYLTRANSFERASE (cyt19)Stephen B. Waters1 , Felicia Walton1 , Miroslav Styblo1 , Karen Herbin-Davis2, and David J. Thomas2 1 School of Medicine, University of North Carolina at Chape...

  7. Increased growth and root Cu accumulation of Sorghum sudanense by endophytic Enterobacter sp. K3-2: Implications for Sorghum sudanense biomass production and phytostabilization.

    Science.gov (United States)

    Li, Ya; Wang, Qi; Wang, Lu; He, Lin-Yan; Sheng, Xia-Fang

    2016-02-01

    Endophytic bacterial strain K3-2 was isolated from the roots of Sorghum sudanense (an bioenergy plant) grown in a Cu mine wasteland soils and characterized. Strain K3-2 was identified as Enterobacter sp. based on 16S rRNA gene sequence analysis. Strain K3-2 exhibited Cu resistance and produced 1-aminocyclopropane-1-carboxylate (ACC) deaminase, indole-3-acetic acid (IAA), siderophores, and arginine decarboxylase. Pot experiments showed that strain K3-2 significantly increased the dry weight and root Cu accumulation of Sorghum sudanense grown in the Cu mine wasteland soils. Furthermore, increase in total Cu uptake (ranging from 49% to 95%) of the bacterial inoculated-Sorghum sudanense was observed compared to the control. Notably, most of Cu (83-86%) was accumulated in the roots of Sorghum sudanense. Furthermore, inoculation with strain K3-2 was found to significantly increase Cu bioconcentration factors and the proportions of IAA- and siderophore-producing bacteria in the root interiors and rhizosphere soils of Sorghum sudanense compared with the control. Significant decrease in the available Cu content was also observed in the rhizosphere soils of the bacterial-inoculated Sorghum sudanense. The results suggest that the endophytic bacterial strain K3-2 may be exploited for promoting Sorghum sudanense biomass production and Cu phytostabilization in the Cu mining wasteland soils. Copyright © 2015 Elsevier Inc. All rights reserved.

  8. Radiation balance in the sweet sorghum crop

    International Nuclear Information System (INIS)

    Assis, F.N. de; Mendez, M.E.G.; Martins, S.R.; Verona, L.A.

    1987-01-01

    The fluxes of incident solar radiation, reflected and net radiation were measured during the growing cicle of two fields of sweet sorghum (Sorghum bicolor L.), cus. BR-501 and BR-503, maintained under convenient irrigation level. Resultant data allowed to estimate the crop albedo as well as the estimates of Rn. (M.A.C.) [pt

  9. Evaluation of sweet sorghum (Sorghum bicolor L. [Moench]) on several population density for bioethanol production

    Science.gov (United States)

    Suwarti; Efendi, R.; Massinai, R.; Pabendon, M. B.

    2018-03-01

    Sweet sorghum (Sorghum bicolor L. [Moench]) crop management that is use for raw source of bioethanol for industrial purpose in Indonesia is less developed. The aim of this research was to evaluated sweet sorghum variety at several population to determine optimum density for juice production. Experiment design was set on split-plot design with three replications, conducted on August to December 2016 at the Indonesian Cereals Research Institute Research Station, Maros South Sulawesi. Main plot were six variation of plant row, and sub plot were three sweet sorghum varieties. Result of the study showed that plant population was high significanty affect to stalk weight, total biomass yield, leaf weight, and also significantly affect bagass weight and juice volume. Varieties were high significantly different in plant height, juice volume, and number of nodes. Super 1 variety on population at 166,667 plants/ha (P1) was obtained the highest juice volume (19,445 lHa-1), meanwhile the highest brix value obtained from Numbu at the same plants population. Furthermore juice volume had significant correlation with biomass weight at the r=0.73. Based on ethanol production, Super 2 and Numbu had the highest volume at 83.333 plants/ha density (P3) and Super 1 at 166.667 plants/ha density with the ethanol volume were 827.68 l Ha-1, 1116.50 l/ha and 993.62 l Ha-1 respectively.

  10. EFFECT OF MECHANICAL CONDITIONING ON THIN-LAYER DRYING OF ENERGY SORGHUM (Sorghum bicolor (L.) Moench)

    Energy Technology Data Exchange (ETDEWEB)

    Ian J. Bonner; Kevin L. Kenney

    2012-10-01

    Cellulosic energy varieties of Sorghum bicolor (L.) Moench show promise as a bioenergy feedstock, however, high moisture content at the time of harvest results in unacceptable levels of degradation when stored in aerobic conditions. To safely store sorghum biomass for extended periods in baled format, the material must be dried to inhibit microbial growth. One possible solution is allowing the material to dry under natural in-field conditions. This study examines the differences in thin-layer drying rates of intact and conditioned sorghum under laboratory-controlled temperatures and relative humidity levels (20 degrees C and 30 degrees C from 40% to 85% relative humidity), and models experimental data using the Page’s Modified equation. The results demonstrate that conditioning drastically accelerates drying times. Relative humidity had a large impact on the time required to reach a safe storage moisture content for intact material (approximately 200 hours at 30 degrees C and 40% relative humidity and 400 hours at 30 degrees C and 70% relative humidity), but little to no impact on the thin-layer drying times of conditioned material (approximately 50 hours for all humidity levels < 70% at 30 degrees C). The drying equation parameters were influenced by temperature, relative humidity, initial moisture content, and material damage, allowing drying curves to be empirically predicted. The results of this study provide valuable information applicable to the agricultural community and to future research on drying simulation and management of energy sorghum.

  11. Genome-Wide Identification and Analysis of Arabidopsis Sodium Proton Antiporter (NHX and Human Sodium Proton Exchanger (NHE Homologs in Sorghum bicolor

    Directory of Open Access Journals (Sweden)

    P. Hima Kumari

    2018-05-01

    Full Text Available Na+ transporters play an important role during salt stress and development. The present study is aimed at genome-wide identification, in silico analysis of sodium-proton antiporter (NHX and sodium-proton exchanger (NHE-type transporters in Sorghum bicolor and their expression patterns under varied abiotic stress conditions. In Sorghum, seven NHX and nine NHE homologs were identified. Amiloride (a known inhibitor of Na+/H+ exchanger activity binding motif was noticed in both types of the transporters. Chromosome 2 was found to be a hotspot region with five sodium transporters. Phylogenetic analysis inferred six ortholog and three paralog groups. To gain an insight into functional divergence of SbNHX/NHE transporters, real-time gene expression was performed under salt, drought, heat, and cold stresses in embryo, root, stem, and leaf tissues. Expression patterns revealed that both SbNHXs and SbNHEs are responsive either to single or multiple abiotic stresses. The predicted protein–protein interaction networks revealed that only SbNHX7 is involved in the calcineurin B-like proteins (CBL- CBL interacting protein kinases (CIPK pathway. The study provides insights into the functional divergence of SbNHX/NHE transporter genes with tissue specific expressions in Sorghum under different abiotic stress conditions.

  12. Changes in protein and starch digestibility in sorghum flour during heat-moisture treatments.

    Science.gov (United States)

    Vu, Thanh-Hien; Bean, Scott; Hsieh, Chao-Feng; Shi, Yong-Cheng

    2017-11-01

    Heat-moisture treatment (HMT) has been used to modify properties of sorghum starches. However, information is limited on the effects of HMT on the digestibility of starch and the concurrent changes in protein in sorghum flour. The objectives of this research were to identify heat-moisture conditions to increase the resistant starch (RS) content of sorghum flour and investigate changes in sorghum proteins and starch structure. Sorghum flours with different moisture contents (0, 125, 200, and 300 g kg -1 w.b.) were heated at three temperatures (100, 120 and 140 °C) and times (1, 2 and 4 h). HMT of sorghum flour increased its RS level. The flour treated at 200 g kg -1 moisture and 100 °C for 4 h had a high RS content (221 g kg -1 vs. 56 g kg -1 for the untreated flour). Starch was not gelatinized when sorghum flours heated at moisture content of 200 g kg -1 or below. Sorghum protein digestibility and solubility decreased during HMT. The increase in RS of sorghum flour upon HMT was attributed to enhanced amylose-lipid complexes and heat induced structural changes in its protein fraction. HMT can be used to increase RS content in sorghum flour without gelatinizing its starch, thereby providing sorghum flour with unique food applications. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  13. Sorghum as a renewable feedstock for production of fuels and industrial chemicals

    Directory of Open Access Journals (Sweden)

    Nhuan P. Nghiem

    2016-01-01

    Full Text Available Considerable efforts have been made in the USA and other countries to develop renewable feedstocks for production of fuels and chemicals. Among these, sorghum has attracted strong interest because of its many good characteristics such as rapid growth and high sugar accumulation, high biomass production potential, excellent nitrogen usage efficiency, wide adaptability, drought resistance, and water lodging tolerance and salinity resistance. The ability to withstand severe drought conditions and its high water usage efficiency make sorghum a good renewable feedstock suitable for cultivation in arid regions, such as the southern US and many areas in Africa and Asia. Sorghum varieties include grain sorghum, sweet sorghum, and biomass sorghum. Grain sorghum, having starch content equivalent to corn, has been considered as a feedstock for ethanol production. Its tannin content, however, may cause problems during enzyme hydrolysis. Sweet sorghum juice contains sucrose, glucose and fructose, which are readily fermentable by Saccharomyces cerevisiae and hence is a good substrate for ethanol fermentation. The enzyme invertase, however, needs to be added to convert sucrose to glucose and fructose if the juice is used for production of industrial chemicals in fermentation processes that employ microorganisms incapable of metabolizing sucrose. Biomass sorghum requires pretreatment prior to enzymatic hydrolysis to generate fermentable sugars to be used in the subsequent fermentation process. This report reviews the current knowledge on bioconversion of sorghum to fuels and chemicals and identifies areas that deserve further studies.

  14. Cloning, expression, crystallization and preliminary X-ray data analysis of norcoclaurine synthase from Thalictrum flavum

    International Nuclear Information System (INIS)

    Pasquo, Alessandra; Bonamore, Alessandra; Franceschini, Stefano; Macone, Alberto; Boffi, Alberto; Ilari, Andrea

    2008-01-01

    The cloning, expression, crystallization and preliminary X-ray data analysis of norcoclaurine synthase from T. flavum, a protein which catalyzes the first committed step in the biosynthesis of benzylisoquinoline alkaloids, are reported. Norcoclaurine synthase (NCS) catalyzes the condensation of 3,4-dihydroxyphenylethylamine (dopamine) and 4-hydroxyphenylacetaldehyde (4-HPAA) as the first committed step in the biosynthesis of benzylisoquinoline alkaloids in plants. The protein was cloned, expressed and purified. Crystals were obtained at 294 K by the hanging-drop vapour-diffusion method using ammonium sulfate and sodium chloride as precipitant agents and diffract to better than 3.0 Å resolution using a synchrotron-radiation source. The crystals belong to the trigonal space group P3 1 21, with unit-cell parameters a = b = 86.31, c = 118.36 Å. A selenomethionine derivative was overexpressed, purified and crystallized in the same space group. A complete MAD data set was collected at 2.7 Å resolution. The model is under construction

  15. [Cloning, Expression and Immunodiagnostic Evaluation of the Fasciola gigantica Thioredoxin Peroxidase].

    Science.gov (United States)

    Wang, Yue-qi; Zhou, Yan; Cheng, Na; Chen, Mu-xin; Ai, Lin; Liu, Yu-hua; Zhang, Jian-guo; Luo, Jia-jun; Xu, Xue-nian

    2015-04-01

    To immunoscreen the gene encoding thioredoxin peroxidase (TPx) from a cDNA library made from adult Fasciola gigantica worms, clone and express the gene, and evaluate the immunodiagnostic value of TPx recombinant protein. The A ZAP cDNA library was immunoscreened with pooled serum of fascioliasis gigantica patients. The obtained positive clones were sequenced and analyzed by multiple sequence alignment. The full-length (rFgTPx) and N-termianal truncated (rFgTPx_nt) sequence of FgTPx was subcloned into prokaryotic plasmid pET28a(+) with a non-fusion expression technique, respectively. The recombinant proteins of rFgTPx and rFgTPx_nt were purified by His-bind affinity column (Ni-NTA). rFgTPx and rFgTPx_nt were used in indirect ELISA to test the antibody response of the serum samples. Sera of 27 fascioliasis gigantica patients, 15 patients with schistosomaisis japonica, 15 clonorchiasis sinensis patients, and 32 healthy donors were tested by using the recombinant protein based ELISA. The TPx recombinant proteins were obtained through expression, purification and renaturation, the relative molecular mass of rFgTPx and rFgTPx_nt were Mr 30,000 and Mr 26,000, respectively. The total diagnostic coincidence rate, sensitivity and specificity of rFgTPx_nt-based ELISA was 87.6% (78/89), 66.7% (18/27), and 96.8% (60/62), respectively. The cross reaction with Schistosoma japonicum and Clonorchis sinensis was 0 and 1/15 for rFgTPx_nt, respectively. Before and after treatment, A450 value of the serum samples from fascioliasis patients was 0.233 ± 0.088 and 0.129 ± 0.072, respectively (t = 4.27, P Fasciola gigantica infection.

  16. Effect of sowing date on grain quality of sorghum ( Sorghum bicolor ...

    African Journals Online (AJOL)

    IVHAA) while minerals; iron and zinc were determined using Atomic Absorption Spectrophotometry. Significant site by variety by sowing date interactions at P < 0.05 level of probability were obtained for protein, iron and zinc content of sorghum ...

  17. Rapid in silico cloning of genes using expressed sequence tags (ESTs).

    Science.gov (United States)

    Gill, R W; Sanseau, P

    2000-01-01

    Expressed sequence tags (ESTs) are short single-pass DNA sequences obtained from either end of cDNA clones. These ESTs are derived from a vast number of cDNA libraries obtained from different species. Human ESTs are the bulk of the data and have been widely used to identify new members of gene families, as markers on the human chromosomes, to discover polymorphism sites and to compare expression patterns in different tissues or pathologies states. Information strategies have been devised to query EST databases. Since most of the analysis is performed with a computer, the term "in silico" strategy has been coined. In this chapter we will review the current status of EST databases, the pros and cons of EST-type data and describe possible strategies to retrieve meaningful information.

  18. Cloning and expression of clt genes encoding milk-clotting proteases from Myxococcus xanthus 422.

    Science.gov (United States)

    Poza, M; Prieto-Alcedo, M; Sieiro, C; Villa, T G

    2004-10-01

    The screening of a gene library of the milk-clotting strain Myxococcus xanthus 422 constructed in Escherichia coli allowed the description of eight positive clones containing 26 open reading frames. Only three of them (cltA, cltB, and cltC) encoded proteins that exhibited intracellular milk-clotting ability in E. coli, Saccharomyces cerevisiae, and Pichia pastoris expression systems.

  19. Efficacy of herbicide seed treatments for controlling Striga infestation of Sorghum

    NARCIS (Netherlands)

    Tuinstra, M.R.; Soumana, S.; Al-Khatib, K.; Kapran, I.; Toure, A.; Ast, van A.; Bastiaans, L.; Ochanda, N.W.; Salami, I.; Kayentao, M.; Dembele, S.

    2009-01-01

    Witchweed (Striga spp.) infestations are the greatest obstacle to sorghum [Sorghum bicolor (L.) Moench] grain production in many areas in Africa. The objective of this study was to evaluate the efficacy of herbicide seed treatments for controlling Striga infestation of sorghum. Seeds of an

  20. Evaluation of sorghum genotypes under drought stress conditions ...

    African Journals Online (AJOL)

    Seven genotypes of sorghum (Sorghum bicolour (L.) Moench) were studied in both drought and normal conditions. In each condition, the genotypes were evaluated using a split plot based randomized complete block design with three replications. Drought tolerance indices including stability tolerance index (STI), mean ...

  1. Performance of Broiler Chicks Fed Irradiated Sorghum Grains

    International Nuclear Information System (INIS)

    Farag, M.D.D.; Farag, M.F. S. El-D.; Afify, A.S.

    2003-01-01

    Substitution of yellow corn with raw sorghum grains in chick diets resulted in decreases in live body weight, accumulative feed consumption and efficiency of feed utilization as compared with reference diet. Relative to raw sorghum diet, inclusion of sorghum grains irradiated at 60 and 100 kGy and/or supplemented with PEG in chick diets resulted in increases in accumulative feed consumption an efficiency feed utilization. The study suggested that irradiation treatment up to 100 kGy up grade broiler chicks performance and the combinations between radiation and PEG treatments sustain the effect of each other

  2. Tapping the US historic sweet sorghum collection to identify biofuel germplasm

    Science.gov (United States)

    Sweet sorghum [Sorghum bicolor (L.) Moench] has gained an important role as a viable alternative to fossil fuels and a more profitable option than maize and sugarcane. Nevertheless, the actual narrow genetic base in sweet sorghum breeding programs is limiting the development of new biofuel varietie...

  3. PAV markers in Sorghum bicolour

    DEFF Research Database (Denmark)

    Shen, Xin; Liu, Zhiquan; Mocoeur, Anne Raymonde Joelle

    2015-01-01

    Abstract Genic presence/absence variants (PAVs) correlate closely to the phenotypic variation, impacting plant genome sizes and the adaption to the environment. To shed more light on their genome-wide patterns, functions and to test the possibility of using them as molecular markers, we analyzed...... enriched in stress responses and protein modification. We used 325 polymorphic PAVs in two sorghum inbred lines Ji2731 and E-Tian, together with 49 SSR markers, and constructed a genetic map, which consisted of 10 linkage groups corresponding to the 10 chromosomes of sorghum and spanned 1430.3 cM in length...

  4. Study on genotypic variation for ethanol production from sweet sorghum juice

    Energy Technology Data Exchange (ETDEWEB)

    Ratnavathi, C.V.; Suresh, K.; Kumar, B.S. Vijay; Pallavi, M.; Komala, V.V.; Seetharama, N. [Directorate of Sorghum Research, Rajendranagar, Hyderabad 500030, Andhra Pradesh (India)

    2010-07-15

    Sugarcane molasses is the main source for ethanol production in India. Sweet sorghum with its juicy stem containing sugars equivalent to that of sugarcane is a very good alternative for bio-ethanol production to meet the energy needs of the country. Sweet sorghum is drought resistant, water logging resistant and saline-alkaline tolerant. Growing sweet sorghum for ethanol production is relatively easy and economical and ethanol produced from sweet sorghum is eco-friendly. In view of this, it is important to identify superior genotypes for ethanol production in terms of percent juice brix, juice extractability, total fermentable sugars, ethanol yield and fermentation efficiency. This paper presents the study on the variability observed for the production of ethanol by various sweet sorghum genotypes in a laboratory fermentor. Five Sweet Sorghum (Sorghum bicolor L. Moench) genotypes were evaluated for ethanol production from stalk juice (Keller, SSV 84, Wray, NSSH 104 and BJ 248). Sweet sorghum juice differs from cane juice mainly in its higher content of starch and aconitic acid. Data were collected for biomass yield; stalk sugar yield and ethanol production in five genotypes. Maximum ethanol production of 9.0%w/v ethanol was obtained with Keller variety (20% sugar concentration was used), and decreased for other genotypes. A distiller's strain of Saccharomyces cerevisiae (gifted by Seagram Distilleries Ltd.) was employed for fermentation. The fermentation efficiency (FE) was 94.7% for this strain. High biomass of yeast was obtained with BJ 248 variety. When the similar experiments were conducted with unsterile sweet sorghum juice (15% sugar concentration) 6.47%w/v ethanol was produced. (author)

  5. Factors influencing beta-amylase activity in sorghum malt

    CSIR Research Space (South Africa)

    Taylor, JRN

    1993-09-01

    Full Text Available isozyme of pI approximately 4.4-4.5, unlike the many isozymes all of higher pI in barley. However, like barley, sorghum beta-amylase was more temperature-labile than its alpha-amylase. Beta-amylase activity in sorghum malt was increased by germination time...

  6. The environment strongly affects estimates of heterosis in hybrid sweet sorghum

    Science.gov (United States)

    Sweet sorghum (Sorghum bicolor (L.) Moench) has potential as a biofuel feedstock but hybrid cultivars are needed to support an industry based on this crop. The purpose of this study was to compare five inbred sweet sorghum lines and 15 hybrids derived from them, and to determine the extent of envir...

  7. Molecular cloning and expression of the IL-10 gene from guinea pigs.

    Science.gov (United States)

    Dirisala, Vijaya R; Jeevan, Amminikutty; Bix, Gregory; Yoshimura, Teizo; McMurray, David N

    2012-04-25

    The Guinea pig (Cavia porcellus) is one of the most relevant small animals for modeling human tuberculosis (TB) in terms of susceptibility to low dose aerosol infection, the organization of granulomas, extrapulmonary dissemination and vaccine-induced protection. It is also considered to be a gold standard for a number of other infectious and non-infectious diseases; however, this animal model has a major disadvantage due to the lack of readily available immunological reagents. In the present study, we successfully cloned a cDNA for the critical Th2 cytokine, interleukin-10 (IL-10), from inbred Strain 2 guinea pigs using the DNA sequence information provided by the genome project. The complete open reading frame (ORF) consists of 537 base pairs which encodes a protein of 179 amino acids. This cDNA sequence exhibited 87% homology with human IL-10. Surprisingly, it showed only 84% homology with the previously published IL-10 sequence from the C4-deficient (C4D) guinea pig, leading us to clone IL-10 cDNA from the Hartley strain of guinea pig. The IL-10 gene from the Hartley strain showed 100% homology with the IL-10 sequence of Strain 2 guinea pigs. In order to validate the only published IL-10 sequence existing in Genbank reported from C4D guinea pigs, genomic DNA was isolated from tissues of C4D guinea pigs. Amplification with various sets of primers showed that the IL-10 sequence reported from C4D guinea pigs contained numerous errors. Hence the IL-10 sequence that is being reported by us replaces the earlier sequence making our IL-10 sequence to be the first one accurate from guinea pig. Recombinant guinea pig IL-10 proteins were subsequently expressed in both prokaryotic and eukaryotic cells, purified and were confirmed by N-terminal sequencing. Polyclonal anti-IL-10 antibodies were generated in rabbits using the recombinant IL-10 protein expressed in this study. Taken together, our results indicate that the DNA sequence information provided by the genome project

  8. Estimation of in situ mating systems in wild sorghum (Sorghum ...

    Indian Academy of Sciences (India)

    The high outcrossing rates of wild/weedy sorghum populations in Ethiopia indicate a high potential for crop genes (including transgenes) to spread within the wild pool. Therefore, effective risk management strategies may be needed if the introgression of transgenes or other crop genes from improved cultivars into wild or ...

  9. cDNA cloning and mRNA expression of cat and dog Cdkal1

    Directory of Open Access Journals (Sweden)

    Sako T

    2012-08-01

    Full Text Available Ichiro Yamamoto, Shingo Ishikawa, Li Gebin, Hiroshi Takemitsu, Megumi Fujiwara, Nobuko Mori, Yutaka Hatano, Tomoko Suzuki, Akihiro Mori, Nobuhiro Nakao, Koh Kawasumi, Toshinori Sako, Toshiro AraiLaboratory of Veterinary Biochemistry, Nippon Veterinary and Life Science University, Tokyo, JapanAbstract: The cyclin-dependent kinase 5 regulatory subunit–associated protein 1–like 1 (CDKAL1 gene encodes methylthiotransferase, and the gene contains risk variants for type 2 diabetes in humans. In this study, we performed complementary DNA cloning for Cdkal1 in the cat and dog and characterized the tissue expression profiles of its messenger RNA. Cat and dog Cdkal1 complementary DNA encoded 576 and 578 amino acids, showing very high sequence homology to mammalian CDKAL1 (>88.4%. Real-time polymerase chain reaction analyses revealed that Cdkal1 messenger RNA is highly expressed in smooth muscle and that tissue distribution of Cdkal1 is similar in cats and dogs. Genotyping analysis of single-nucleotide polymorphism for cat Cdkal1 revealed that obese cats had different tendencies from normal cats. These findings suggest that the cat and dog Cdkal1 gene is highly conserved among mammals and that cat Cdkal1 may be a candidate marker for genetic diagnosis of obesity.Keywords: cat, dog, Cdkal1, obese, cDNA cloning, Q-PCR

  10. [Cloning, prokaryotic expression and antibacterial assay of Tenecin gene encoding an antibacterial peptide from Tenebrio molitor].

    Science.gov (United States)

    Liu, Ying; Jiang, Yu-xin; Li, Chao-pin

    2011-12-01

    To clone tenecin gene, an antibacterial peptide gene, from Tenebrio molitor for its prokaryotic expression and explore the molecular mechanism for regulating the expression of antibacterial peptide in Tenebrio molitor larvae. The antibacterial peptide was induced from the larvae of Tenebrio molitor by intraperitoneal injection of Escherichia coli DH-5α (1×10(8)/ml). RT-PCR was performed 72 h after the injection to clone Tenecin gene followed by sequencing and bioinformatic analysis. The recombinant expression vector pET-28a(+)-Tenecin was constructed and transformed into E. coli BL21(DE3) cells and the expression of tenecin protein was observed after IPTG induction. Tenecin expression was detected in transformed E.coli using SDS-PAGE after 1 mmol/L IPTG induction. Tenecin gene, which was about 255 bp in length, encoded Tenecin protein with a relative molecular mass of 9 kD. Incubation of E.coli with 80, 60, 40, and 20 µg/ml tenecin for 18 h resulted in a diameter of the inhibition zone of 25.1∓0.03, 20.7∓0.06, 17.2∓0.11 and 9.3∓0.04 mm, respectively. Tenecin protein possesses strong antibacterial activity against E. coli DH-5α, which warrants further study of this protein for its potential as an antibacterial agent in clinical application.

  11. Lactic acid fermentation of two sorghum varieties is not affected by ...

    African Journals Online (AJOL)

    The study was conducted to investigate sorghum grain variety differences in lactic acid fermentation based on their differences in phenolic contents. The study wa s conductedas a 2 x 5 x 4 factorial design with three factors: Factor 1: Sorghum variety (white and red sorghum); Factor 2: Control treatment without lactic acid ...

  12. Epicoccum nigrum the new pathogen of sorghum seed in Serbia

    Directory of Open Access Journals (Sweden)

    Ristić Danijela

    2012-01-01

    Full Text Available Sixteen samples of sorghum seed (Sorghum bicolor (L. Moench. 'Alba', 'Gold', 'Prima' and 'Reform' were analyzed in the localities of Bački Petrovac and Čantavir in the period 2009-2011. Tipresence of species belonging to the genera Epicoccum, Fusarium, Alternaria, Aspergillus and Penicillium was established in single and mixed infections. From the infected sorghum seed, monosporial cultures identified as Epicoccum nigrum based on morphology, proved their pathogenicity on artificially inoculated sorghum seedlings. Molecular identification was performed by PCR and amplification of the ITS region of ribosomal DNA. Gene sequences of selected isolates 291-09 (JQ619838 and 315-09 (JQ619839 exhibited 99-100% nucleotide identity with the sequences of 31 isolates of E. nigrum deposited in the GenBank. It obtained results represent the first detailed characterization of E. nigrum in Serbia. The presence of a large number of phytopathogenic fungi on sorghum seed should be further investigated in order to clarify their relationships and relative significance.

  13. Incorporating a Sorghum Habitat for Enhancing Lady Beetles (Coleoptera: Coccinellidae in Cotton

    Directory of Open Access Journals (Sweden)

    P. G. Tillman

    2012-01-01

    Full Text Available Lady beetles (Coleoptera: Coccinellidae prey on insect pests in cotton. The objective of this 2 yr on-farm study was to document the impact of a grain sorghum trap crop on the density of Coccinellidae on nearby cotton. Scymnus spp., Coccinella septempunctata (L., Hippodamia convergens Guérin-Méneville, Harmonia axyridis (Pallas, Coleomegilla maculata (De Geer, Cycloneda munda (Say, and Olla v-nigrum (Mulsant were found in sorghum over both years. Lady beetle compositions in sorghum and cotton and in yellow pyramidal traps were similar. For both years, density of lady beetles generally was higher on cotton with sorghum than on control cotton. Our results indicate that sorghum was a source of lady beetles in cotton, and thus incorporation of a sorghum habitat in farmscapes with cotton has great potential to enhance biocontrol of insect pests in cotton.

  14. Economic feasibility of producing sweet sorghum as an ethanol feedstock in the southeastern United States

    International Nuclear Information System (INIS)

    Linton, Joseph A.; Miller, J. Corey; Little, Randall D.; Petrolia, Daniel R.; Coble, Keith H.

    2011-01-01

    This study examines the feasibility of producing sweet sorghum (Sorghum bicolor (L.) Moench) as an ethanol feedstock in the southeastern United States through representative counties in Mississippi. We construct enterprise budgets along with estimates of transportation costs to estimate sweet sorghum producers' breakeven costs for producing and delivering sweet sorghum biomass. This breakeven cost for the sweet sorghum producer is used to estimate breakeven costs for the ethanol producer based on wholesale ethanol price, production costs, and transportation and marketing costs. Stochastic models are developed to estimate profits for sweet sorghum and competing crops in two representative counties in Mississippi, with sweet sorghum consistently yielding losses in both counties. -- Highlights: → We examine the economic feasibility of sweet sorghum as an ethanol feedstock. → We construct enterprise budgets along with estimates of transportation costs. → We estimate breakeven costs for producing and delivering sweet sorghum biomass. → Stochastic models determine profits for sweet sorghum in two Mississippi counties.

  15. Cloning and expression analysis of a novel ammonium transporter gene from eichhornia

    International Nuclear Information System (INIS)

    Li, Y.; Yan, G.; Zheng, L.

    2014-01-01

    In order to explore the molecular mechanism for Eichhornia crassipes to transport ammonium from outside, we cloned a novel ammonium transporter (EcAMT) gene from E. crassipes and identified its function by using yeast complementation experiment. The full-length cDNA of EcAMT contains a 1506 nucletide-long open reading frame which encodes a protein of 501 amino acids. Bioinformatics analysis predicted that EcAMT had 8 transmembrane regions. The expressions of EcAMT gene under three different nitrogen conditions were evaluated by quantitative reverse transcriptase PCR (qRT-PCR) and the results showed that the expression of EcAMT gene was up-regulated under nitrogen starvation. Our study results revealed some molecular mechanism of E. crassipes to absorb the ammonium in eutrophic water. (author)

  16. PHYTOCHEMICAL STUDY OF A TINCTORIAL PLANT OF BENIN TRADITIONAL PHARMACOPOEIA: THE RED SORGHUM (Sorghum caudatum) OF BENIN

    OpenAIRE

    PASCAL D. C. AGBANGNAN; CHRISTINE TACHON; HELENE BONIN; ANNA CHROSTOWKA; ERIC FOUQUET; DOMINIQUE C. K. SOHOUNHLOUE

    2012-01-01

    The full phytochemical screening of red sorghum from Benin (Sorghum caudatum) achieved in this work reveals the presence of leucoanthocyanins, flavonoides, free quinones, combined anthracene derivatives, sterols and terpenes in higher concentration in the leaf sheath and marrow of stem than in the seed. Catechin tannin content is 11.4% in the leaf sheath (slightly higher than that of red wine), 5.8% in the marrow and 2.8% in the seed. Gallic tannins, saponins and the mucilage present in the l...

  17. Grain sorghum is a viable feedstock for ethanol production.

    Science.gov (United States)

    Wang, D; Bean, S; McLaren, J; Seib, P; Madl, R; Tuinstra, M; Shi, Y; Lenz, M; Wu, X; Zhao, R

    2008-05-01

    Sorghum is a major cereal crop in the USA. However, sorghum has been underutilized as a renewable feedstock for bioenergy. The goal of this research was to improve the bioconversion efficiency for biofuels and biobased products from processed sorghum. The main focus was to understand the relationship among "genetics-structure-function-conversion" and the key factors impacting ethanol production, as well as to develop an energy life cycle analysis model (ELCAM) to quantify and prioritize the saving potential from factors identified in this research. Genetic lines with extremely high and low ethanol fermentation efficiency and some specific attributes that may be manipulated to improve the bioconversion rate of sorghum were identified. In general, ethanol yield increased as starch content increased. However, no linear relationship between starch content and fermentation efficiency was found. Key factors affecting the ethanol fermentation efficiency of sorghum include protein digestibility, level of extractable proteins, protein and starch interaction, mash viscosity, amount of phenolic compounds, ratio of amylose to amylopectin, and formation of amylose-lipid complexes in the mash. A platform ELCAM with a base case showed a positive net energy value (NEV) = 25,500 Btu/gal EtOH. ELCAM cases were used to identify factors that most impact sorghum use. For example, a yield increase of 40 bu/ac resulted in NEV increasing from 7 million to 12 million Btu/ac. An 8% increase in starch provided an incremental 1.2 million Btu/ac.

  18. Characterizing Sorghum Panicles using 3D Point Clouds

    Science.gov (United States)

    Lonesome, M.; Popescu, S. C.; Horne, D. W.; Pugh, N. A.; Rooney, W.

    2017-12-01

    To address demands of population growth and impacts of global climate change, plant breeders must increase crop yield through genetic improvement. However, plant phenotyping, the characterization of a plant's physical attributes, remains a primary bottleneck in modern crop improvement programs. 3D point clouds generated from terrestrial laser scanning (TLS) and unmanned aerial systems (UAS) based structure from motion (SfM) are a promising data source to increase the efficiency of screening plant material in breeding programs. This study develops and evaluates methods for characterizing sorghum (Sorghum bicolor) panicles (heads) in field plots from both TLS and UAS-based SfM point clouds. The TLS point cloud over experimental sorghum field at Texas A&M farm in Burleston County TX were collected using a FARO Focus X330 3D laser scanner. SfM point cloud was generated from UAS imagery captured using a Phantom 3 Professional UAS at 10m altitude and 85% image overlap. The panicle detection method applies point cloud reflectance, height and point density attributes characteristic of sorghum panicles to detect them and estimate their dimensions (panicle length and width) through image classification and clustering procedures. We compare the derived panicle counts and panicle sizes with field-based and manually digitized measurements in selected plots and study the strengths and limitations of each data source for sorghum panicle characterization.

  19. Sorghum as an alternative of cultivation to maize; Sorghumhirse als Anbaualternative zum Mais

    Energy Technology Data Exchange (ETDEWEB)

    Jaekel, Kerstin; Theiss, Markus; Poetzschke, Karen [Saechsisches Landesamt fuer Umwelt, Landwirtschaft und Geologie (LfULG), Dresden (Germany)] [and others

    2013-10-01

    Due to their high dry matter yield potential Sorghum bicolor and Sorghum bicolor x sudanense are well fitted as feedstock for biogas production. Similar to maize, both species show a high efficiency in their use of water (C4-plants). However, Sorghum has a higher drought tolerance in comparison with maize but is more sensitive to low temperatures. Hence a cultivation of Sorghum is recommendable especially in dry and relatively warm regions, including recultivated areas and even on loess soil, provided that the required temperatures are given. Due to the fact that Sorghum is not affected by the corn root worm, it also could gain relevance in regions were the cultivation of maize is restricted. Furthermore, Sorghum is usable as a catch crop as well as a main crop because of its variable sowing time. Catch crop cultivation, however, yields a significantly lower amount of dry matter and -quality which is a result of its shorter vegetation period. Owing to its higher crude fiber concentration Sorghum achieves a lower theoretically attainable specific methane yield (Weissbach) than maize. Thus only on rare occasions Sorghum does achieve methane yields per hectare that are comparable to maize. Eventually, the competitiveness of Sorghum greatly depends on provision of enhanced cultivars achieved through genetic improvement. (orig.)

  20. Cloning and expression of the receptor for human urokinase plasminogen activator, a central molecule in cell surface, plasmin dependent proteolysis

    DEFF Research Database (Denmark)

    Roldan, A.L.; Cubellis, M.V.; Masucci, M.T.

    1990-01-01

    , and therefore the capacity of cells to migrate and invade neighboring tissues. We have isolated a 1.4 kb cDNA clone coding for the entire human uPAR. An oligonucleotide synthesized on the basis of the N-terminal sequence of the purified protein was used to screen a cDNA library made from SV40 transformed human......, a size very close to that of the cloned cDNA. Expression of the uPAR cDNA in mouse cells confirms that the clone is complete and expresses a functional uPA binding protein, located on the cell surface and with properties similar to the human uPAR. Caseinolytic plaque assay, immunofluorescence analysis......The surface receptor for urokinase plasminogen activator (uPAR) has been recognized in recent years as a key molecule in regulating plasminogen mediated extracellular proteolysis. Surface plasminogen activation controls the connections between cells, basement membrane and extracellular matrix...

  1. Effects of main traits of sweet sorghum irradiated by carbon ions

    International Nuclear Information System (INIS)

    Li Wenjian; He Jingyu; Liu Qingfang; Yu Lixia; Dong Xicun

    2009-01-01

    To investigate the influence of carbon ion irradiation on important agronomic characters of sweet sorghum, dry seeds of Sweet Sorghum BJ0601 and BJ0602 were irradiated by 100 MeV/u 12 C +6 ion beam to different doses at Heavy Ion Accelerator National Laboratory in Lanzhou (HIANLL). When matured, the main traits of sweet sorghum were measured. The correlation coefficient of five main agronomic characters, i.e. number of node, plant height, stalk diameter, sugar content and stem weight per plant, were analyzed using the SPSS 13.0 software. The results indicated that the obvious influence of sweet sorghum irradiated by carbon ion beam was observed. In addition, the correlation of main traits was studied. This study may provide rudimental data to select novel variety of sweet sorghum suited for fuel ethanol production. In addition, the average of sugar content of early mutant BJ0601-1 is higher than BJ0601 in M2, and the sugar content of sweet sorghum may be improved by carbon ion beam irradiation. (authors)

  2. Mycobacterium tuberculosis HspX/EsxS Fusion Protein: Gene Cloning, Protein Expression, and Purification in Escherichia coli.

    Science.gov (United States)

    Khademi, Farzad; Yousefi-Avarvand, Arshid; Derakhshan, Mohammad; Meshkat, Zahra; Tafaghodi, Mohsen; Ghazvini, Kiarash; Aryan, Ehsan; Sankian, Mojtaba

    2017-10-01

    The purpose of this study was to clone, express, and purify a novel multidomain fusion protein of Micobacterium tuberculosis (Mtb) in a prokaryotic system. An hspX/esxS gene construct was synthesized and ligated into a pGH plasmid, E. coli TOP10 cells were transformed, and the vector was purified. The vector containing the construct and pET-21b (+) plasmid were digested with the same enzymes and the construct was ligated into pET-21b (+). The accuracy of cloning was confirmed by colony PCR and sequencing. E. coli BL21 cells were transformed with the pET-21b (+)/hspX/esxS expression vector and protein expression was evaluated. Finally, the expressed fusion protein was purified on a Ni-IDA column and verified by SDS-PAGE and western blotting. The hspX/esxS gene construct was inserted into pET-21b (+) and recombinant protein expression was induced with IPTG in E. coli BL21 cells. Various concentrations of IPTG were tested to determine the optimum concentration for expression induction. The recombinant protein was expressed in insoluble inclusion bodies. Three molar guanidine HCl was used to solubilize the insoluble protein. An HspX/EsxS Mtb fusion protein was expressed in E. coli and the recombinant protein was purified. After immunological analysis, the HspX/EsxS fusion protein might be an anti-tuberculosis vaccine candidate in future clinical trial studies.

  3. Molecular cloning and expression in Saccharomyces cerevisiae and Neurospora crassa of the invertase gene from Neurospora crassa.

    Science.gov (United States)

    Carú, M; Cifuentes, V; Pincheira, G; Jiménez, A

    1989-10-01

    A plasmid (named pCN2) carrying a 7.6 kb BamHI DNA insert was isolated from a Neurospora crassa genomic library raised in the yeast vector YRp7. Saccharomyces cerevisiae suco and N. crassa inv strains transformed with pNC2 were able to grow on sucrose-based media and expressed invertase activity. Saccharomyces cerevisiae suco (pNC2) expressed a product which immunoreacted with antibody raised against purified invertase from wild type N. crassa, although S. cerevisiae suc+ did not. The cloned DNA hybridized with a 7.6 kb DNA fragment from BamHI-restricted wild type N. crassa DNA. Plasmid pNC2 transformed N. crassa Inv- to Inv+ by integration either near to the endogenous inv locus (40% events) or at other genomic sites (60% events). It appears therefore that the cloned DNA piece encodes the N. crassa invertase enzyme. A 3.8 kb XhoI DNA fragment, derived from pNC2, inserted in YRp7, in both orientation, was able to express invertase activity in yeast, suggesting that it contains an intact invertase gene which is not expressed from a vector promoter.

  4. Effects of nitrogen fertilizer application and solar radiation on the growth response of sorghum [Sorghum bicolor] seedlings to soil moisture

    International Nuclear Information System (INIS)

    Sumi, A.; Katayama, T.C.

    2000-01-01

    The effects of nitrogen fertilizer application and solar radiation on the growth response to soil moisture were examined in sorghum seedlings grown in culture boxes. The effects of soil moisture (f) and amount of nitrogen fertilizer application (g) on the increment of total dry matter weight of sorghum seedling (ΔW) were represented satisfactorily by the following reciprocal equation, 1/ΔW = A/(f - f 0 ) + B(g + g 0 )/(f - f 0 ) + C/[(f - f 0 ) (g + g 0 )] + D/(g + g 0 ) + E, where f 0 and g 0 were the uppermost value of unavailable soil moisture and the amount of nitrogen supplied from soil and seeds. A, B, C, D and E were coefficients. The effects of soil moisture (f) and solar radiation (S) on ΔW were expressed approximately by the following reciprocal equation, 1/ΔW = A/(S - S 0 ) + B/(f - f 0 ) + C(f - f 0 ) + D, where S 0 was the daily compensation point. These results indicated that the effects of solar radiation and soil moisture are additive, but the interaction between soil moisture and nitrogen fertilizer is not negligible. The transpiration efficiency was unaffected by soil moisture, nitrogen fertilizer and solar radiation

  5. PHYTOCHEMICAL STUDY OF A TINCTORIAL PLANT OF BENIN TRADITIONAL PHARMACOPOEIA: THE RED SORGHUM (Sorghum caudatum OF BENIN

    Directory of Open Access Journals (Sweden)

    PASCAL D. C. AGBANGNAN

    2012-06-01

    Full Text Available The full phytochemical screening of red sorghum from Benin (Sorghum caudatum achieved in this work reveals the presence of leucoanthocyanins, flavonoides, free quinones, combined anthracene derivatives, sterols and terpenes in higher concentration in the leaf sheath and marrow of stem than in the seed. Catechin tannin content is 11.4% in the leaf sheath (slightly higher than that of red wine, 5.8% in the marrow and 2.8% in the seed. Gallic tannins, saponins and the mucilage present in the leaf sheath and marrow, are virtually absent in the seed. Marrow and leaf sheath extracts (1 g/50 mL showed a concentration of anthocyanins (147 mg/L and 213.5 mg/L similar to that of rosy wine and red wine with short maceration. The grain of sorghum is four times, respectively two times less rich in phenolic compounds than the leaf sheath and the marrow of stem.

  6. Cloning the interleukin 1 receptor from human T cells

    International Nuclear Information System (INIS)

    Sims, J.E.; Acres, R.B.; Grubin, C.E.; McMahan, C.J.; Wignall, J.M.; March, C.J.; Dower, S.K.

    1989-01-01

    cDNA clones of the interleukin 1 (IL-1) receptor expressed in a human T-cell clone have been isolated by using a murine IL-1 receptor cDNA as a probe. The human and mouse receptors show a high degree of sequence conservation. Both are integral membrane proteins possessing a single membrane-spanning segment. Similar to the mouse receptor, the human IL-1 receptor contains a large cytoplasmic region and an extracellular, IL-1 binding portion composed of three immunoglobulin-like domains. When transfected into COS cells, the human IL-1 receptor cDNA clone leads to expression of two different affinity classes of receptors, with K a values indistinguishable from those determined for IL-1 receptors in the original T-cell clone. An IL-1 receptor expressed in human dermal fibroblasts has also been cloned and sequenced and found to be identical to the IL-1 receptor expressed in T cells

  7. A versatile and efficient high-throughput cloning tool for structural biology.

    Science.gov (United States)

    Geertsma, Eric R; Dutzler, Raimund

    2011-04-19

    Methods for the cloning of large numbers of open reading frames into expression vectors are of critical importance for challenging structural biology projects. Here we describe a system termed fragment exchange (FX) cloning that facilitates the high-throughput generation of expression constructs. The method is based on a class IIS restriction enzyme and negative selection markers. FX cloning combines attractive features of established recombination- and ligation-independent cloning methods: It allows the straightforward transfer of an open reading frame into a variety of expression vectors and is highly efficient and very economic in its use. In addition, FX cloning avoids the common but undesirable feature of significantly extending target open reading frames with cloning related sequences, as it leaves a minimal seam of only a single extra amino acid to either side of the protein. The method has proven to be very robust and suitable for all common pro- and eukaryotic expression systems. It considerably speeds up the generation of expression constructs compared to traditional methods and thus facilitates a broader expression screening.

  8. Complementation of radiation-sensitive Ataxia telangiectasia cells after transfection of cDNA expression libraries and cosmid clones from wildtype cells

    International Nuclear Information System (INIS)

    Fritz, E.

    1994-06-01

    In this Ph.D.-thesis, phenotypic complementation of AT-cells (AT5BIVA) by transfection of cDNA-expression-libraries was adressed: After stable transfection of cDNA-expression-libraries G418 resistant clones were selected for enhanced radioresistance by a fractionated X-ray selection. One surviving transfectant clone (clone 514) exhibited enhanced radiation resistance in dose-response experiments and further X-ray selections. Cell cycle analysis revealed complementation of untreated and irradiated 514-cells in cell cycle progression. The rate of DNA synthesis, however, is not diminished after irradiation but shows the reverse effect. A transfected cDNA-fragment (AT500-cDNA) was isolated from the genomic DNA of 514-cells and proved to be an unknown DNA sequence. A homologous sequence could be detected in genomic DNA from human cell lines, but not in DNA from other species. The cDNA-sequence could be localized to human chromosome 11. In human cells the cDNA sequence is part of two large mRNAs. 4 different cosmid clones containing high molecular genomic DNA from normal human cells could be isolated from a library, each hybridizing to the AT500-cDNA. After stable transfection into AT-cells, one cosmid-clone was able to confer enhanced radiation resistance both in X-ray selections and dose-response experiments. The results indicate that the cloned cDNA-fragment is based on an unknown gene from human chromosome 11 which partially complements the radiosensitivity and the defective cell cycle progression in AT5BIVA cells. (orig.) [de

  9. Mutation breeding in sorghum (sorghum bicolor L.) for improving plant as ruminant feed

    International Nuclear Information System (INIS)

    H, Soeranto

    1998-01-01

    Mutation breeding using gamma irradiation in sorghum was aimed at improving the quality and production of sorghum plant as ruminant feed. Seeds of local sorghum variety Keris with moisture of about 14% were irradiated with gamma rays from Cobalt-60 source using the dose levels up to 0.5 kgy. The MI plant were grown in Pasar Jumat, the M2 and M3 were grown in Citayam experimental station. The M2 plants were harvested 40 days after sowing by cutting plants 20 cm above ground surface. Two weeks later observations for the ability of plants to produce new buds (buds variable). The plants green products in green products in from of their dry weight (product variable) were collected 40 days after harvesting and drying process in oven at 105 0 C for 24 hours. Plant selections with intensity of 20% were done for the bud variable among samples of M2 plants. Selection responses in the M3 were found to vary from the lowest at 0.5 kgy population (R s = 0.8507). The share of genetic factors to selection responses in bud variable varied from 7.25% at 0,5 kgy population to 22.35% at 0.3 kgy population. Selection for bud variable gave directly impact in increasing product variable in the M3. (author)

  10. Sorghum bi-color

    African Journals Online (AJOL)

    sunny

    2014-11-12

    Nov 12, 2014 ... Biomass materials require reduction and densification for the purpose of handling and space requirements. Guinea corn (Sorghum bi-color) is a major source of biomass material in the tropic regions. The densification process involves some ... a closed-end die, the temperature and the use of binder.

  11. Cloning and expression of a human kidney cDNA for an α2-adrenergic receptor subtype

    International Nuclear Information System (INIS)

    Regan, J.W.; Kobilka, T.S.; Yang-Feng, T.L.; Caron, M.G.; Lefkowitz, R.J.; Kobilka, B.K.

    1988-01-01

    An α 2 -adrenergic receptor subtype has been cloned from a human kidney cDNA library using the gene for the human platelet α 2 -adrenergic receptor as a probe. The deduced amino acid sequence resembles the human platelet α 2 -adrenergic receptor and is consistent with the structure of other members of he family of guanine nucleotide-binding protein-coupled receptors. The cDNA was expressed in a mammalian cell line (COS-7), and the α 2 -adrenergic ligand [ 3 H]rauwolscine was bound. Competition curve analysis with a variety of adrenergic ligands suggests that this cDNA clone represents the α 2 B-adrenergic receptor. The gene for this receptor is on human chromosome 4, whereas the gene for the human platelet α 2 -adrenergic receptor (α 2 A) lies on chromosome 10. This ability to express the receptor in mammalian cells, free of other adrenergic receptor subtypes, should help in developing more selective α-adrenergic ligands

  12. Molecular cloning and expression analysis of three omega-6 desaturase genes from purslane (Portulaca oleracea L.).

    Science.gov (United States)

    Teixeira, M C; Coelho, N; Olsson, M E; Brodelius, P E; Carvalho, I S; Brodelius, M

    2009-07-01

    Two full-length cDNA clones of PoleFAD2 and one full-length cDNA clone of PoleFAD6, encoding omega-6 fatty acid desaturases, the key enzymes for the conversion of oleic into linoleic acid, were isolated from purslane (Portulaca oleracea L.) leaves and seeds. The deduced amino acid sequence of both isoforms of PoleFAD2 showed higher similarities to other microsomal omega-6 desaturases then to PoleFAD6 or other plastidial orthologues, and vice versa. Expression analysis by RT-PCR showed that all genes are expressed in all tissues of purslane tested, but higher levels of mRNA accumulation were detected in reproductive organs and cells that proliferate rapidly or store lipids. Wounding affected the levels of mRNA accumulation of both, FAD2 and FAD6 genes in purslane leaves, while chilling stress affected only FAD2 transcript level. The expression patterns observed reflect the discrete roles of these genes in membrane synthesis for cell division, thylakoid development, and lipid storage or in the biosynthetic pathway for the production of signaling molecules that influence plant development or defense.

  13. Molecular cloning and expression of the gene encoding the kinetoplast-associated type II DNA topoisomerase of Crithidia fasciculata.

    Science.gov (United States)

    Pasion, S G; Hines, J C; Aebersold, R; Ray, D S

    1992-01-01

    A type II DNA topoisomerase, topoIImt, was shown previously to be associated with the kinetoplast DNA of the trypanosomatid Crithidia fasciculata. The gene encoding this kinetoplast-associated topoisomerase has been cloned by immunological screening of a Crithidia genomic expression library with monoclonal antibodies raised against the purified enzyme. The gene CfaTOP2 is a single copy gene and is expressed as a 4.8-kb polyadenylated transcript. The nucleotide sequence of CfaTOP2 has been determined and encodes a predicted polypeptide of 1239 amino acids with a molecular mass of 138,445. The identification of the cloned gene is supported by immunoblot analysis of the beta-galactosidase-CfaTOP2 fusion protein expressed in Escherichia coli and by analysis of tryptic peptide sequences derived from purified topoIImt. CfaTOP2 shares significant homology with nuclear type II DNA topoisomerases of other eukaryotes suggesting that in Crithidia both nuclear and mitochondrial forms of topoisomerase II are encoded by the same gene.

  14. Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of the regulator AcrR from Escherichia coli

    International Nuclear Information System (INIS)

    Li, Ming; Qiu, Xi; Su, Chih-Chia; Long, Feng; Gu, Ruoyu; McDermott, Gerry; Yu, Edward W.

    2006-01-01

    The transcriptional regulator AcrR from Escherichia coli has been cloned, overexpressed, purified and crystallized and X-ray diffraction data have been collected to a resolution of 2.5 Å. This paper describes the cloning, expression, purification and preliminary X-ray data analysis of the AcrR regulatory protein. The Escherichia coli AcrR is a member of the TetR family of transcriptional regulators. It regulates the expression of the AcrAB multidrug transporter. Recombinant AcrR with a 6×His tag at the C-terminus was expressed in E. coli and purified by metal-affinity chromatography. The protein was crystallized using hanging-drop vapor diffusion. X-ray diffraction data were collected from cryocooled crystals at a synchrotron light source. The best crystal diffracted to 2.5 Å. The space group was determined to be P3 2 , with unit-cell parameters a = b = 46.61, c = 166.16 Å

  15. NDVI to Detect Sugarcane Aphid Injury to Grain Sorghum.

    Science.gov (United States)

    Elliott, N C; Backoulou, G F; Brewer, M J; Giles, K L

    2015-06-01

    Multispectral remote sensing has potential to provide quick and inexpensive information on sugarcane aphid, Melanaphis sacchari (Zehntner), pest status in sorghum fields. We describe a study conducted to determine if injury caused by sugarcane aphid to sorghum plants in fields of grain sorghum could be detected using multispectral remote sensing from a fixed wing aircraft. A study was conducted in commercial grain sorghum fields in the Texas Gulf Coast region in June 2014. Twenty-six commercial grain sorghum fields were selected and rated for the level of injury to sorghum plants in the field caused by sugarcane aphid. Plant growth stage ranged from 5.0 (watery ripe) to 7.0 (hard dough) among fields; and plant injury rating from sugarcane aphid ranged from 1.0 (little or no injury) to 4.0 (>40% of plants displaying injury) among fields. The normalized differenced vegetation index (NDVI) is calculated from light reflectance in the red and near-infrared wavelength bands in multispectral imagery and is a common index of plant stress. High NDVI indicates low levels of stress and low NDVI indicates high stress. NDVI ranged from -0.07 to 0.26 among fields. The correlation between NDVI and plant injury rating was negative and significant, as was the correlation between NDVI and plant growth stage. The negative correlation of NDVI with injury rating indicated that plant stress increased with increasing plant injury. Reduced NDVI with increasing plant growth probably resulted from reduced photosynthetic activity in more mature plants. The correlation between plant injury rating and plant growth stage was positive and significant indicating that plant injury from sugarcane aphid increased as plants matured. The partial correlation of NDVI with plant injury rating was negative and significant indicating that NDVI decreased with increasing plant injury after adjusting for its association with plant growth stage. We demonstrated that remotely sensed imagery acquired from grain

  16. Electrochemical evaluation of sweet sorghum fermentable sugar bioenergy feedstock

    Science.gov (United States)

    Redox active constituents of sorghum, e.g., anthocyanin, flavonoids, and aconitic acid, putatively contribute to its pest resistance. Electrochemical reactivity of sweet sorghum stem juice was evaluated using cyclic voltammetry (CV) for five male (Atlas, Chinese, Dale, Isidomba, N98) and three fema...

  17. Utilization of Iles-Iles and Sorghum Starch for Bioethanol Production

    Directory of Open Access Journals (Sweden)

    Kusmiyati Kusmiyati

    2014-05-01

    Full Text Available The aims of this study were to convert the starches from iles-iles tubers (Amorphophalus campanulatus and sorghum grains (Sorghum bicolor L into bioethanol as an alternative energy. Both of these agricultural products contains a high content starches and they do not use as the major foods in Indonesia. To find out the maximum ethanol concentration and yield, both the raw materials were converted to ethanol on various process variables including the concentration of flour substrate solution (100-300 g/L, β-amylase enzyme concentration (0.8 - 6.4 ml/kg of flour , the  concentration of dry yeast S. cerevisiae (2-15 g, and fermentation time (72-168 hours. The results showed that at the flour substrate concentration of 250 g/L produced the maximum ethanol contents of 100.29 g/L and 95.11 g/L   for iles-iles and sorghum, respectively. Effect of β-amylase enzyme in the saccharification process showed that at concentration  of 3.2 ml/kg  the maximum reducing sugar content of 204.94 g/L and 193.15 g/L  for iles-iles and sorghum substrate, respectively were generated therefore it was corresponding to the maximum ethanol production. The concentration effect of dry yeast S. cerevisiae in the fermentation stage for the iles-iles and sorghum substrate revealed that the maximum ethanol obtained at 5 g yeast activated in 100 ml medium starter resulted the highest ethanol content 100.29 g/L 95.11 g/L for iles-iles and sorghum substrate, respectively. To determine the effect of fermentation time on ethanol yield from iles-iles and sorghum substrate, the fermentation process were performed at 3, 5, and 7 days. The maximum ethanol fermentation was obtained at 5 days fermentation. The ethanol yield is calculated by weight of ethanol is formed (g divided by the weight of flour (g. Based on the experiment results, conducted, generally the highest ethanol yield of iles-iles was higher than that of sorghum flour. The highest yield (g/g iles-iles and sorghum

  18. Cloning of the chrysanthemum UEP1 promoter and comparative expression in leaves and ray and disc florets of Dendranthema grandiflora

    NARCIS (Netherlands)

    Annadana, S.; Beekwilder, M.J.; Kuipers, G.; Visser, P.B.; Outchkourov, N.; Pereira, A.; Udayakumar, M.; Jongsma, M.A.

    2002-01-01

    To attain high transgene expression in petal tissue of ray florets of chrysanthemum an endogenous ubiquitin extension protein (UEP1) promoter was cloned and tested with the β-glucuronidase (GUS) reporter gene. Expression levels were compared with four heterologous promoters: chalcone synthase

  19. Performance of elite grain sorghum varieties in the West Nile Agro ...

    African Journals Online (AJOL)

    ACSS

    Ouma, J.P. and Akuja, T.E. 2013. Agronomic and morphological performance of sorghum (Sorghum bicolor L.) for the dry highlands of. Kenya. www.m.elewa.org. Schatz, B.G., Schneiter, A.A. and Gardner,. J.E. 1987. Effect of plant density on grain sorghum production in North. Dakota. pp. 16-17. Snider, J.L., Randy, L.R. and ...

  20. Fungal endophytes of sorghum in Burkina Faso

    DEFF Research Database (Denmark)

    Zida, E P; Thio, I G; Néya, B J

    2014-01-01

    A survey was conducted to assess the natural occurrence and distribution of fungal endophytes in sorghum in relation to plant performance in two distinct agro-ecological zones in Burkina Faso. Sorghum farm-saved seeds were sown in 48 farmers’ fields in Sahelian and North Sudanian zones to produce...... sorghum plants. In each field, leaf samples were collected from five well-developed (performing) and five less-developed (non-performing) plants at 3-5 leaf stage, while at plant maturity leaf, stem and root samples were collected from the same plants and fungal endophytes were isolated. A total of 39...... fungal species belonging to 25 genera were isolated. The most represented genera included Fusarium, Leptosphaeria, Curvularia, Nigrospora and Penicillium. The genera Fusarium and Penicillium occurred significantly higher in performing plants as compared to non-performing plants while the genera...

  1. Experimental study on bread yeast cultured in sweet sorghum juice

    International Nuclear Information System (INIS)

    Wang Jufang; Dong Xicun; Li Wenjian; Xiao Guoqing; Ma Liang; Gao Feng

    2008-01-01

    As a substitute for food supplies, sweet sorghum juice with high grade has demonstrated out- standing advantage in fermentation. To obtain the optimized fermentation conditions, the growth, the bio- mass of bread yeast cultured in sweet sorghum juice and total residual sugar were investigated in the paper. The fermentation was performed and optimized in a 10-100 1 bio-reactor. The results show that the application of sweet sorghum juice in bread yeast production is very potential. (authors)

  2. Sorghum stem yield and soluble carbohydrates under different ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-11-19

    Nov 19, 2008 ... The aim of this study was to select the most suitable cultivar for salty land in this geographical area. Two sweet sorghum cultivars (Keller and Sofra) and one grain sorghum cultivar (Kimia) were grown in greenhouse benches under four salinity levels of 2, 4, 8 and 12 dSm-1 to evaluate the effects of salinity.

  3. Morphological characteristics of BRS 501 sweet sorghum under water stress

    Directory of Open Access Journals (Sweden)

    Luciano Rezende Moreira

    2016-12-01

    Full Text Available Sorghum [Sorghum bicolor (L. Moench] crop is distinguished from other crops for its tolerance to both water deficit and excess soil moisture, under very dry and/or very hot environmental situations in which the productivity of other cereals becomes uneconomical. This work was conducted to evaluate the effects of irrigation on root conformation at the initial development phase of sweet sorghum. So, BRS 501 cv. was subjected to four irrigation levels based on 80%, 60%, 40% and 20% of the field capacity (CC. The decreased availability of water in the soil negatively affected the majority of the characteristics under evaluation except for the relationship between the root system and the aerial part (SR/PA, average root diameter (DMR and specific root area (ARE. We concluded that the growth of sweet sorghum plants under evaluation is sensible to the decrease of water in the soil, as it is affected by low water availability. This methodology, common to other crops, can be used for saccharine sorghum in order to establish hydric availabilities in new experiments to discriminate the drought-tolerant cultivars.

  4. Cloning, expression, purification, crystallization and initial crystallographic analysis of the preprotein translocation ATPase SecA from Thermus thermophilus

    International Nuclear Information System (INIS)

    Vassylyeva, Marina N.; Mori, Hiroyuki; Tsukazaki, Tomoya; Yokoyama, Shigeyuki; Tahirov, Tahir H.; Ito, Koreaki; Vassylyev, Dmitry G.

    2006-01-01

    The SecA ATPase from T. thermophilus was cloned, expressed, purified and crystallized. Complete diffraction data sets were collected for two crystal forms at 2.8 and 3.5 Å resolution, respectively. Determination of the structure is now in progress. The Thermus thermophilus gene encoding the preprotein translocation ATPase SecA was cloned and expressed and the purified protein was crystallized by the hanging-drop vapour-diffusion technique in two different space groups P3 1(2) 21 (a = b = 168.6, c = 149.8 Å) and P6 1(5) 22 (a = b = 130.9, c = 564.6 Å). The crystals, improved by macroseeding, diffracted to beyond 2.8 and 3.5 Å resolution for the trigonal and hexagonal crystal forms, respectively. Structure determination using the multiple isomorphous replacement method is in progress

  5. The Dermatophagoides farinae group 22 allergen: cloning and expression in Escherichia coli.

    Science.gov (United States)

    Cui, Yu-bao; Cai, Hong-xing; Zhou, Ying; Wang, Nan; Yu, Li-li; Yang, Li; Zhang, Cheng-bo

    2015-09-01

    Dermatophagoides farinae (Hughes) (Acari: Pyroglyphidae) and other domestic mites produce allergens that affect people worldwide. Here, the complementary DNA (cDNA) coding for group 22 allergen of D. farinae (Der f 22) from China was cloned, sequenced, and expressed successfully. The cDNA encoding Der f 22 was synthesized by reverse transcription polymerase chain reaction (RT-PCR), then ligated to the pCold-TF for expression in Escherichia coli BL21 cells. The purified recombinant fusion protein was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), Western-blotting, and tandem matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF/TOF). The full-length cDNA comprised 468 nucleotides and was 99.57% (466/468) identical with the reference sequence (GenBank: DQ643992). After the plasmid pCold-TF-Der f 22 was transformed into E. coli BL21 and expressed with the induction of IPTG, SDS-PAGE showed a specific band for the recombinant fusion protein. The recombinant fusion protein, which was purified by chromatography, bound with a His-tagged antibody by Western blotting. MALDI-TOF/TOF mass spectrometry revealed that the structure of the recombinant protein was identical to the predicted Der f 22 structure. The hydrophilic protein contains a signal peptide of 20 amino acids, and the mature Der f 22 consists of 135 amino acid residues with a molecular weight of 14.7 kDa and theoretical isoelectric points (pI) of 6.38. Its secondary structure comprises an alpha helix (38.5%), beta-sheet (45.9%), random coils (11.85%), and beta-turns (11.1%). This work represents the first reported full-length sequence and successful cloning of Der f 22 from D. farinae in China; bioinformatics analysis can be used to further study the allergenicity and clinical utility of the recombinant Der f 22. © 2015 ARS-AAOA, LLC.

  6. Cloning, expression, and purification of recombinant protein MPT-64 from a virulent strain of Mycobacterium bovis in a prokaryotic system.

    Science.gov (United States)

    Tashakkori, Maryam Mohammadi; Tebianian, Majid; Tabatabaei, Mohammad; Mosavari, Nader

    2016-12-01

    Tuberculosis (TB) is a zoonotic infectious disease common to humans and animals that is caused by the rod-shaped acid-fast bacterium Mycobacterium bovis. Rapid and sensitive detection of TB is promoted by specific antigens. Virulent strains of the TB complex from M. bovis contain 16 regions of difference (RD) in their genome that encode important proteins, including major protein of Mycobacterium Tuberculosis 64 (MBT-64, which is a primary immune-stimulating antigen encoded by RD-2. In this study, we cloned, expressed, and purified MPT-64 as a potent M. bovis antigen in a prokaryotic system for use in future diagnostic studies. The antigenic region of the Mpt64 gene was investigated by bioinformatics methods, cloned into the PQE-30 plasmid, and expressed in Escherichia coli M15 cells, followed by isopropyl β-d-1-thiogalactopyranoside induction. The expressed protein was analyzed sodium dodecyl sulfate polyacrylamide gel electrophoresis and purified using a nickel-affinity column. Biological activity was confirmed by western blot using specific antibodies. Our data verified the successful cloning of the Mpt64 gene (687-bp segment) via the expression vector and purification of recombinant MPT-64 as a 24-kDa protein. These results indicated successful expression and purification of recombinant MPT-64 protein in a prokaryotic system. This protein can be used for serological diagnosis, improved detection of pathogenicity and non-pathogenicity between infected cattle, and for verification of suspected cases of bovine TB. Copyright © 2016.

  7. Factors That Influence Technical Efficiency of Sorghum Production: A Case of Small Holder Sorghum Producers in Lower Eastern Kenya

    Directory of Open Access Journals (Sweden)

    Evaline Chepng’etich

    2015-01-01

    Full Text Available Majority of the rural households in Kenya depend on agriculture as a source of food and livelihood. Agricultural productivity has been declining due to many factors resulting in increased food insecurity in the country. Consequently, there is a renewed interest in promoting drought-tolerant crops such as sorghum which thrives in the arid and semiarid lands of the developing world. However, performance of sorghum production among the smallholder farmers has still remained low. This study was thus carried out to identify factors that influence technical efficiency of sorghum production among smallholder farmers in Machakos and Makindu districts of the lower eastern Kenya. Collected data on farm and farmer characteristics were analysed by use of descriptive statistics and Tobit model. Result highlights show that technical efficiency was influenced positively by formal education level of the household, experience in sorghum farming, membership in farmers associations, use of hired labour, production advice, and use of manure. Surprisingly household size, meant to enhance labour, had a negative influence. To increase technical efficiency, efforts should focus on improving information flows on agronomic practices. Farmers should also be encouraged to form and actively participate in various farmers associations, which enhance learning and pooling of labour resources, hence improving technical efficiency.

  8. Energy sorghum--a genetic model for the design of C4 grass bioenergy crops.

    Science.gov (United States)

    Mullet, John; Morishige, Daryl; McCormick, Ryan; Truong, Sandra; Hilley, Josie; McKinley, Brian; Anderson, Robert; Olson, Sara N; Rooney, William

    2014-07-01

    Sorghum is emerging as an excellent genetic model for the design of C4 grass bioenergy crops. Annual energy Sorghum hybrids also serve as a source of biomass for bioenergy production. Elucidation of Sorghum's flowering time gene regulatory network, and identification of complementary alleles for photoperiod sensitivity, enabled large-scale generation of energy Sorghum hybrids for testing and commercial use. Energy Sorghum hybrids with long vegetative growth phases were found to accumulate more than twice as much biomass as grain Sorghum, owing to extended growing seasons, greater light interception, and higher radiation use efficiency. High biomass yield, efficient nitrogen recycling, and preferential accumulation of stem biomass with low nitrogen content contributed to energy Sorghum's elevated nitrogen use efficiency. Sorghum's integrated genetics-genomics-breeding platform, diverse germplasm, and the opportunity for annual testing of new genetic designs in controlled environments and in multiple field locations is aiding fundamental discovery, and accelerating the improvement of biomass yield and optimization of composition for biofuels production. Recent advances in wide hybridization between Sorghum and other C4 grasses could allow the deployment of improved genetic designs of annual energy Sorghums in the form of wide-hybrid perennial crops. The current trajectory of energy Sorghum genetic improvement indicates that it will be possible to sustainably produce biofuels from C4 grass bioenergy crops that are cost competitive with petroleum-based transportation fuels. © The Author 2014. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  9. [Cloning and expressing of cyclophilin B gene from Schistosoma japonnicum and the analysis of immunoprotective effect].

    Science.gov (United States)

    Peng, Jinbiao; Han, Hongxiao; Hong, Yang; Wang, Yan; Guo, Fanji; Shi, Yaojun; Fu, Zhiqiang; Liu, Jinming; Cheng, Guofeng; Lin, Jiaojiao

    2010-03-01

    The present study was intend to clone and express the cDNA encoding Cyclophilin B (CyPB) of Schistosoma japonicum, its preliminary biological function and further immunoprotective effect against schistosome infection in mice. RT-PCR technique was applied to amplify a full-length cDNA encoding protein Cyclophilin B (Sj CyPB) from schistosomula cDNA. The expression profiles of Sj CyPB were determined by Real-time PCR using the template cDNAs isolated from 7, 13, 18, 23, 32 and 42 days parasites. The cDNA containing the Open Reading Frame of CyPB was then subcloned into a pGEX-6P-1 vector and transformed into competent Escherichia coli BL21 for expressing. The recombinant protein was renaturated, purified and its antigenicity were detected by Western blotting, and the immunoprotective effect induced by recombinant Sj CyPB was evaluated in Balb/C mice. The cDNA containing the ORF of Sj CyPB was cloned with the length of 672 base pairs, encoding 223 amino acids. Real-time PCR analysis revealed that the gene had the highest expression in 18-day schistosomula, suggesting that Sj CyPB was schistosomula differentially expressed gene. The recombinant protein showed a good antigenicity detected by Western blotting. Animal experiment indicated that the vaccination of recombinant CyPB protein in mice led to 31.5% worm and 41.01% liver egg burden reduction, respectively, compared with those of the control. A full-length cDNA differentially expressed in schistosomula was obtained. The recombinant Sj CyPB protein could induce partial protection against schistosome infection.

  10. Cloning, expression, purification, crystallization and X-ray analysis of inositol monophosphatase from Mus musculus and Homo sapiens

    International Nuclear Information System (INIS)

    Singh, Nisha; Halliday, Amy C.; Knight, Matthew; Lack, Nathan A.; Lowe, Edward; Churchill, Grant C.

    2012-01-01

    M. musculus and H. sapiens inositol monophosphatase 1 were cloned, expressed, purified and crystallized. Diffraction data were collected and analysed at resolutions of 2.4 and 1.7 Å, respectively, and the structures were compared in order to identify any structural differences. Inositol monophosphatase (IMPase) catalyses the hydrolysis of inositol monophosphate to inositol and is crucial in the phosphatidylinositol (PI) signalling pathway. Lithium, which is the drug of choice for bipolar disorder, inhibits IMPase at therapeutically relevant plasma concentrations. Both mouse IMPase 1 (MmIMPase 1) and human IMPase 1 (HsIMPase 1) were cloned into pRSET5a, expressed in Escherichia coli, purified and crystallized using the sitting-drop method. The structures were solved at resolutions of 2.4 and 1.7 Å, respectively. Comparison of MmIMPase 1 and HsIMPase 1 revealed a core r.m.s. deviation of 0.516 Å

  11. Survival of Skin Graft between Transgenic Cloned Dogs and Non-Transgenic Cloned Dogs

    Science.gov (United States)

    Kim, Geon A; Oh, Hyun Ju; Kim, Min Jung; Jo, Young Kwang; Choi, Jin; Park, Jung Eun; Park, Eun Jung; Lim, Sang Hyun; Yoon, Byung Il; Kang, Sung Keun; Jang, Goo; Lee, Byeong Chun

    2014-01-01

    Whereas it has been assumed that genetically modified tissues or cells derived from somatic cell nuclear transfer (SCNT) should be accepted by a host of the same species, their immune compatibility has not been extensively explored. To identify acceptance of SCNT-derived cells or tissues, skin grafts were performed between cloned dogs that were identical except for their mitochondrial DNA (mtDNA) haplotypes and foreign gene. We showed here that differences in mtDNA haplotypes and genetic modification did not elicit immune responses in these dogs: 1) skin tissues from genetically-modified cloned dogs were successfully transplanted into genetically-modified cloned dogs with different mtDNA haplotype under three successive grafts over 63 days; and 2) non-transgenic cloned tissues were accepted into transgenic cloned syngeneic recipients with different mtDNA haplotypes and vice versa under two successive grafts over 63 days. In addition, expression of the inserted gene was maintained, being functional without eliciting graft rejection. In conclusion, these results show that transplanting genetically-modified tissues into normal, syngeneic or genetically-modified recipient dogs with different mtDNA haplotypes do not elicit skin graft rejection or affect expression of the inserted gene. Therefore, therapeutically valuable tissue derived from SCNT with genetic modification might be used safely in clinical applications for patients with diseased tissues. PMID:25372489

  12. Sorghum - An alternative energy crop for marginal lands and reclamation sites

    Science.gov (United States)

    Lukas, Stefan; Theiß, Markus; Jäkel, Kerstin

    2017-04-01

    The production of biogas and the associated cultivation of energy crops are still of great importance. Considering increasing restrictions for the cultivation of standard biogas crop maize regarding an environmentally friendly production of biomass, a wider range of energy crops is needed. The cultivation of sorghum can contribute to this. As maize, sorghum is a C4-plant and offers a high biomass yield potential. Originated in the semi-arid tropics, sorghum is well adapted to warm and dry climate and particularly noted for its drought tolerance compared to maize. It also makes few demands on soil quality and shows a good capability of nutrient acquisition. Therefore, particularly on marginal areas and reclamation sites with low soil nutrient and water content sorghum can contribute to secure crop yield and income of farmers. The applied research project aims at and reflects on the establishment of sorghum as a profitable and ecological friendly cropping alternative to maize, especially in the face of probable climate change with increasing risks for agriculture. For this purpose, site differentiated growing and cultivar trials with a standardized planting design as well as several practical on-farm field experiments were conducted. The agronomical and economic results will lead to scientifically based procedures and standards for agricultural practice with respect to cultivation methods (drilling, pest-management, fertilization), cropping sequence and technique, cropping period or position in crop rotation. Even by now there is a promising feedback from the agricultural practice linked with an increasing demand for information. Moreover, the specific cropping area is increasing continuously. Therefore, the leading signs for the establishment of sorghum as profitable alternative to maize biogas production are positive. Sorghum cultures perform best as main crops in the warm D locations in the middle and East German dry areas. Here, the contribution margin

  13. Antimicrobial evaluation of red, phytoalexin-rich sorghum food biocolorant

    NARCIS (Netherlands)

    Akogou, Folachodé U.G.; Besten, Den Heidy M.W.; Polycarpe Kayodé, A.P.; Fogliano, Vincenzo; Linnemann, Anita R.

    2018-01-01

    Sorghum (Sorghum bicolor) extract is traditionally used as red biocolorant in West Africa to colour foods, among which wagashi, a soft cheese. This biocolorant is a source of the phytoalexin apigeninidin and phenolic acids, and users claim that it has preservative effects next to its colouring

  14. The Effect of Salicylic Acid and Gibberellin on Seed Reserve Utilization, Germination and Enzyme Activity of Sorghum (Sorghum bicolor L. Seeds Under Drought Stress

    Directory of Open Access Journals (Sweden)

    Roghayyeh Sheykhbaglou

    2014-03-01

    Full Text Available Seed priming methods have been used to increases germination characteristics under stress conditions. The study aimed was to determine the effect of salicylic acid and gibberellin on seed reserve utilization, germination and enzyme activity of sorghum (Sorghum bicolor L. seeds under drought stress. Factorial experiment was carried out in completely randomized design with three replications. The first factor was the seed treatments (unpriming, salicylic acid and gibberellin and the second factor was drought stress (0, -4, -8 and -12 bar. The results indicated that for these traits: germination percentage, germination index, weight of utilized (mobilized seed, seed reserve utilization efficiency, seedling dry weight and seed reserve depletion percentage was a significant treatment Ч drought interaction. Thus priming improved study traits in Sorghum (Sorghum bicolor L. seeds under drought stress. Also, priming improves enzyme activity as compared to the unprimed seeds.

  15. Cloning and expression analysis of FaPR-1 gene in strawberry

    Science.gov (United States)

    Mo, Fan; Luo, Ya; Ge, Cong; Mo, Qin; Ling, Yajie; Luo, Shu; Tang, Haoru

    2018-04-01

    The FaPR-1 gene was cloned by RT-PCR from `Benihoppe' strawberry and its bioinformatics analysis was conducted. The results showed that the open reading frame was 483 bp encoding encoding l60 amino acids which protein molecular weight and theoretical isoelectricity were 17854.17 and 8.72 respectively. Subcellular localization prediction shows that this gene is located extracellularly. By comparing strawberry FaPR-l and other plant Pathogenesis-related protein, homology and phylogenetic tree construction showed that the homology with grapes, peach is relatively close. In the treatments of ABA, sucrose and the mixture of the two, the expression of FaPR-1 in strawberry fruit were significantly increased.

  16. Karakteristik Sensori dan Fisiko-Kimia Beras Analog Sorghum dengan Penambahan Rempah Campuran

    Directory of Open Access Journals (Sweden)

    Maya Indra Rasyid

    2017-02-01

    Full Text Available The purpose of this study was to obtain the formula of sorghum rice analogue by mixed spices addition with acceptable sensory and physico-chemical characteristics.  The selection of sorghum rice analogue formula was tested by using hedonic test with 70 untrained panelists. The addition of mixed spices powder was as follows: 30 % onion, 20 % garlic, 10 % bay leaves, 20 % ginger and 20 % lemongrass. Those mixed spices powder were added to the sorghum rice analogue at percentage of 0.25 %, 0,5 %, 1 %, 2 %, 3 % and 0 % (control  from total dough weight. The sorghum rice analogue was made using extrusion technology (a twin screw extruder. The overall sensory evaluation result showed that the addition of spice mixed had significant effect (p ≤ 0.05 on the characteristics of sorghum rice analogue. The panelists accepted the sorghum rice analogue with 1% mixed spice. The preferred formulation was the addition of 1% mixed spice which contain of  9.56 % moisture, 0.72 % ash, 0.53 % fat, 6.22 % protein, 92.53 % carbohydrate, 26.48 % amyloseand 6,67 % dietary fiber. Sorghum rice analogue enriched by spices is a potential as a rich fiber source. ABSTRAK Tujuan penelitian adalah mendapatkan formula beras analog berbahan dasar sorgum dengan penambahan rempah campuran yang dapat diterima secara sensori. Pemilihan formula dilakukan dengan uji hedonik menggunakan 70 orang panelis tidak terlatih. Rempah yang ditambahkan berupa bubuk rempah campuran yang terdiri atas bawang merah 30%, bawang putih 20 %, daun salam 10 %, jahe 20 % dan sereh 20 %. Penambahan bubuk rempah campuran untuk pembuatan beras analog sorghum berturut-turut 0,25 %, 0,5 %, 1 %, 2 %, 3 % dan kontrol (tanpa rempah dari total berat adonan diluar air. Beras analog sorghum dibuat dengan teknologi ekstrusi menggunakan ekstruder ulir ganda. Hasil uji sensori secara keseluruhan menunjukkan bahwa penambahan rempah campuran berpengaruh nyata (p <0,05 terhadap nasi beras analog yang dihasilkan

  17. Registration of six grain sorghum pollinator (R) lines

    Science.gov (United States)

    Six sorghum [Sorghum bicolor (L.) Moench] pollinators [KS142R (Reg. No. PI XXXX), KS143R (Reg. No. PI XXXX), KS144R (Reg.No. PI XXXX), KS145R (Reg. No. PI XXXX), KS146R (Reg. No. PI XXXX) and KS147R (Reg. No. PI XXXX) were developed from random mating using a recurrent selection followed by pedigree...

  18. Effect of Excessive Soil Moisture Stress on Sweet Sorghum: Physiological Changes and Productivity

    International Nuclear Information System (INIS)

    Zhang, F.; Wang, Y.; Yu, H.; Zhu, K.; Zhang, Z.; Zou, F. L. J.

    2016-01-01

    Sweet sorghum [Sorghum bicolor (L.) Moench] is a potential bioenergy feedstock. Research explaining the response of sweet sorghum to excessive soil moisture (EM) stress at different growth stage is limited. To investigate the effect of EM stress on sweet sorghum antioxidant enzymes, osmotic regulation, biomass, quality, and ethanol production, an experiment was conducted in a glasshouse at the National Sorghum Improvement Center, Shenyang, China. Sweet sorghum (cv. LiaoTian1) was studied in four irrigation treatments with a randomized block design method. The results showed that the protective enzyme, particularly the SOD, CAT and APX in it, was significantly affected by EM stress. EM stress deleteriously affected sweet sorghum growth, resulting in a remarkable reduction of aboveground biomass, stalk juice quality, stalk juice yield, and thus, decreased ethanol yield. EM stress also caused significant reduction in plant relative water content, which further decreased stalk juice extraction rate. Sweet sorghum grown under light, medium, and heavy EM treatments displayed 5, 19, and 30% fresh stalk yield reduction, which showed a significant difference compared to control. The estimated juice ethanol yield significantly declined from 1407 ha/sup -1/ (under optimum soil moisture) to 1272, 970, and 734 L ha/sup -1/ respectively. (author)

  19. Effect of gamma irradiation on chemical composition and nutritive value of sorghum grains

    International Nuclear Information System (INIS)

    Mekkawy, S.H.

    1996-01-01

    Sorghum grains were gamma irradiated at 0, 10, 50, 100, 150 and 200 KGy doses using cobalt-60 source. Irradiated and unirradiated sorghum samples were analyzed for crude fiber contents, total nitrogen, fat, ash and tannic acid. Neutral-detergent fiber (NDF), acid-detergent fiber (ADF) and acid detergent lignin (ADL) were also determined. In addition, digestibility coefficient received special attention. The irradiated sorghum grains were incorporated into basal diets and fed to rats during the digestion trials. The results indicated that gamma irradiation had no effects on total nitrogen, fat and ash contents of sorghum grains. Irradiation treatments of sorghum did not cause a pronounced effect on tannic acid content even those received the highest irradiation dose (200 kGy). Moreover, the irradiation treatments decreased the NDF content of sorghum especially those subjected to 100 or 200 kGy. On the other hand, the ADF and ADL values did not show a remarkable change due to irradiation treatments. Hemicellulose content was decreased with the increase of irradiation dose levels. Also, it was noticed that feeding rats on basal diets enriched with irradiated sorghum grains had a beneficial effects on digestibility coefficient. This trend was obvious with animals supplemented with sorghum grains subjected to the relatively high irradiation dose levels. 4 tabs

  20. Predominant lactic acid bacteria associated with the traditional malting of sorghum grains

    DEFF Research Database (Denmark)

    Sawadogo-Lingani, H.; Diawara, B.; Glover, R.K.

    2010-01-01

    dominated the microbiota from sorghum grains to malted sorghum. These isolates had technological properties comparable to those responsible for the acidification of sorghum beer (dolo, pito) wort produced from sorghum malt (previously studied), suggesting their potential for use as starter cultures....... Suitable isolates of L. fermentum are promising candidates to be used as starter cultures from the initial step of malting, that is, the steeping and are expected to inhibit the growth and survival of pathogens and spoilage microflora, and to control the lactic fermentation of dolo and pito wort or other...

  1. Cloning and Expression of Leptospira LipL32 Antigen as a Candidate for Rapid Diagnosis

    Directory of Open Access Journals (Sweden)

    Nooshin Sohrabi

    2013-09-01

    Full Text Available Background and Objective: Leptospirosis as an important emerging infectious zoonotic disease caused by spirochetes of the genus Leptospira. Given the low sensitivity and long duration of its culture, the diagnosis of leptospirosis is mainly based on serological methods. The microscopic agglutination test (MAT is considered as the reference method. Because of the complexity of the MAT, there is an urgent need for the development of new reliable and rapid screening tests for leptospirosis. Major leptospiral outer membrane proteins (OMPs, present only in pathologic strains, could be regarded as a good candidate for diagnostic studies. Here we report the cloning and expression of LipL32, as a prominent immunogenic protein, in a prokaryotic system. Materials and Methods: After the amplification of LipL32 gene, it was cloned into the pQE30 vector. The insertion of LipL32 gene into the vector was screened and confirmed with restriction analysis and sequencing. The recombinant plasmid was transformed into E. coli M15 strain, and the expressed protein was identified by SDS-PAGE and western blotting. This recombinant protein with 6× His-tagged sequence was purified using Ni-NTA affinity column chromatography. Results: The results revealed that the selected gene was successfully cloned in pQE30 vector and recombinant protein (rLipL32 of approximately ~32 kDa was produced, purified and confirmed by western blotting. Conclusion: This recombinant protein could be potentially used for the development of serodiagnosis tests for the diagnosis of leptospirosis in humans and animals.

  2. Performance evaluation of biomass sorghum in Hawaii and Texas

    Science.gov (United States)

    Although biomass sorghum [Sorghum bicolor (L.) Moench] has been identified as a high yielding bioenergy feedstock crop on the continental USA, there is lack of conclusive data on its performance in HI. The objective of this study was to (i) determine the adaptability and productivity of two biomass...

  3. Effect of Sources and Storage Conditions on Quality of Sorghum ...

    African Journals Online (AJOL)

    The germination test of sorghum seeds varied highly significantly (P<0.001) from Kwimba. 74%, Chamwino .... Mean separation test was done using Least. Significance ... for QDS sorghum is 98%. One dot represents more than one sample.

  4. Characteristics of African traditional beers brewed with sorghum malt: a review

    Directory of Open Access Journals (Sweden)

    Lyumugabe, F.

    2012-01-01

    Full Text Available Traditional sorghum beers are produced in several countries of Africa, but variations in the manufacturing process may occur depending on the geographic localization. These beers are very rich in calories, B-group vitamins including thiamine, folic acid, riboflavin and nicotinic acid, and essential amino acids such as lysine. However, the traditional sorghum beer is less attractive than Western beers because of its poorer hygienic quality, organoleptic variations and shorter shelf life. Research into the microbiological and biochemical characteristics of traditional sorghum beers as well as their technologies have been performed and documented in several African countries. This review aims to summarize the production processes and compositional characteristics of African traditional sorghum beers (ikigage, merissa, doro, dolo, pito, amgba and tchoukoutou. It also highlights the major differences between these traditional beers and barley malt beer, consumed worldwide, and suggests adaptations that could be made to improve the production process of traditional sorghum beer.

  5. Genomic dissection of anthracnose resistant response in sorghum [Sorghum bicolor (L.)

    Science.gov (United States)

    The goal of this project is to use a genomics-based approaches to identify anthracnose resistance loci from diverse sorghum germplasm as an effort to the disease resistance mechanism of at least one of these genes. This information will provide plant breeders with a tool kit that can be used to maxi...

  6. Prospecting for Energy-Rich Renewable Raw Materials: Sorghum Stem Case Study.

    Science.gov (United States)

    Byrt, Caitlin S; Betts, Natalie S; Tan, Hwei-Ting; Lim, Wai Li; Ermawar, Riksfardini A; Nguyen, Hai Yen; Shirley, Neil J; Lahnstein, Jelle; Corbin, Kendall; Fincher, Geoffrey B; Knauf, Vic; Burton, Rachel A

    2016-01-01

    Sorghum vegetative tissues are becoming increasingly important for biofuel production. The composition of sorghum stem tissues is influenced by genotype, environment and photoperiod sensitivity, and varies widely between varieties and also between different stem tissues (outer rind vs inner pith). Here, the amount of cellulose, (1,3;1,4)-β-glucan, arabinose and xylose in the stems of twelve diverse sorghum varieties, including four photoperiod-sensitive varieties, was measured. At maturity, most photoperiod-insensitive lines had 1% w/w (1,3;1,4)-β-glucan in stem pith tissue whilst photoperiod-sensitive varieties remained in a vegetative stage and accumulated up to 6% w/w (1,3;1,4)-β-glucan in the same tissue. Three sorghum lines were chosen for further study: a cultivated grain variety (Sorghum bicolor BTx623), a sweet variety (S. bicolor Rio) and a photoperiod-sensitive wild line (S. bicolor ssp. verticilliflorum Arun). The Arun line accumulated 5.5% w/w (1,3;1,4)-β-glucan and had higher SbCslF6 and SbCslH3 transcript levels in pith tissues than did photoperiod-insensitive varieties Rio and BTx623 (<1% w/w pith (1,3;1,4)-β-glucan). To assess the digestibility of the three varieties, stem tissue was treated with either hydrolytic enzymes or dilute acid and the release of fermentable glucose was determined. Despite having the highest lignin content, Arun yielded significantly more glucose than the other varieties, and theoretical calculation of ethanol yields was 10 344 L ha-1 from this sorghum stem tissue. These data indicate that sorghum stem (1,3;1,4)-β-glucan content may have a significant effect on digestibility and bioethanol yields. This information opens new avenues of research to generate sorghum lines optimised for biofuel production.

  7. Prospecting for Energy-Rich Renewable Raw Materials: Sorghum Stem Case Study.

    Directory of Open Access Journals (Sweden)

    Caitlin S Byrt

    Full Text Available Sorghum vegetative tissues are becoming increasingly important for biofuel production. The composition of sorghum stem tissues is influenced by genotype, environment and photoperiod sensitivity, and varies widely between varieties and also between different stem tissues (outer rind vs inner pith. Here, the amount of cellulose, (1,3;1,4-β-glucan, arabinose and xylose in the stems of twelve diverse sorghum varieties, including four photoperiod-sensitive varieties, was measured. At maturity, most photoperiod-insensitive lines had 1% w/w (1,3;1,4-β-glucan in stem pith tissue whilst photoperiod-sensitive varieties remained in a vegetative stage and accumulated up to 6% w/w (1,3;1,4-β-glucan in the same tissue. Three sorghum lines were chosen for further study: a cultivated grain variety (Sorghum bicolor BTx623, a sweet variety (S. bicolor Rio and a photoperiod-sensitive wild line (S. bicolor ssp. verticilliflorum Arun. The Arun line accumulated 5.5% w/w (1,3;1,4-β-glucan and had higher SbCslF6 and SbCslH3 transcript levels in pith tissues than did photoperiod-insensitive varieties Rio and BTx623 (<1% w/w pith (1,3;1,4-β-glucan. To assess the digestibility of the three varieties, stem tissue was treated with either hydrolytic enzymes or dilute acid and the release of fermentable glucose was determined. Despite having the highest lignin content, Arun yielded significantly more glucose than the other varieties, and theoretical calculation of ethanol yields was 10 344 L ha-1 from this sorghum stem tissue. These data indicate that sorghum stem (1,3;1,4-β-glucan content may have a significant effect on digestibility and bioethanol yields. This information opens new avenues of research to generate sorghum lines optimised for biofuel production.

  8. 76 FR 314 - Sorghum Promotion, Research, and Information Program: Referendum

    Science.gov (United States)

    2011-01-04

    ... DEPARTMENT OF AGRICULTURE Agricultural Marketing Service [Doc. No. AMS-LS-10-0103] Sorghum Promotion, Research, and Information Program: Referendum AGENCY: Agricultural Marketing Service, USDA. ACTION: Notice of Opportunity to Participate in the Sorghum Promotion, Research, and Information...

  9. Factors Influencing the Adoption of Improved Sorghum Varieties in ...

    African Journals Online (AJOL)

    The findings of the study indicated that age and distance to input market were negatively and significantly related to improved sorghum varieties whereas farm size and type of house owned were found to have been positively and significantly related to improved sorghum varieties. The results of the study confirm that ...

  10. Overexpression of sweet sorghum cryptochrome 1a confers hypersensitivity to blue light, abscisic acid and salinity in Arabidopsis.

    Science.gov (United States)

    Zhou, Tingting; Meng, Lingyang; Ma, Yue; Liu, Qing; Zhang, Yunyun; Yang, Zhenming; Yang, Deguang; Bian, Mingdi

    2018-02-01

    This work provides the bioinformatics, expression pattern and functional analyses of cryptochrome 1a from sweet sorghum (SbCRY1a), together with an exploration of the signaling mechanism mediated by SbCRY1a. Sweet sorghum [Sorghum bicolor (L.) Moench] is considered to be an ideal candidate for biofuel production due to its high efficiency of photosynthesis and the ability to maintain yield under harsh environmental conditions. Blue light receptor cryptochromes regulate multiple aspects of plant growth and development. Here, we reported the function and signal mechanism of sweet sorghum cryptochrome 1a (SbCRY1a) to explore its potential for genetic improvement of sweet sorghum varieties. SbCRY1a transcripts experienced almost 24 h diurnal cycling; however, its protein abundance showed no oscillation. Overexpression of SbCRY1a in Arabidopsis rescued the phenotype of cry1 mutant in a blue light-specific manner and regulated HY5 accumulation under blue light. SbCRY1a protein was present in both nucleus and cytoplasm. The photoexcited SbCRY1a interacted directly with a putative RING E3 ubiquitin ligase constitutive photomorphogenesis 1 (COP1) from sweet sorghum (SbCOP1) instead of SbSPA1 to suppress SbCOP1-SbHY5 interaction responding to blue light. These observations indicate that the function and signaling mechanism of cryptochromes are basically conservative between monocotyledons and dicotyledons. Moreover, SbCRY1a-overexpressed transgenic Arabidopsis showed oversensitive to abscisic acid (ABA) and salinity. The ABA-responsive gene ABI5 was up-regulated evidently in SbCRY1a transgenic lines, suggesting that SbCRY1a might regulate ABA signaling through the HY5-ABI5 regulon.

  11. Millet and corn oil in sorghum-based diets for broilers

    Directory of Open Access Journals (Sweden)

    João Paulo Rodrigues Bueno

    2015-12-01

    Full Text Available ABSTRACT: This study evaluated the effects of millet and corn oil additions to sorghum-based diets on the performance, carcass yields and prime cuts (i.e., wings, breasts, thighs and drumsticks and the relative weights of edible offal (i.e., gizzard, heart, and liver of broiler chickens. A total of 684 Hubbard Flex chickens, including 342 broilers of each sex, were housed. The design was completely randomized, and the following diets were supplied: A sorghum and soybean meal + soybean oil (control; B sorghum and soybean meal + corn oil; and C sorghum and soybean meal + millet and soybean oil. Six replicates with 38 birds each (19 males and 19 females were evaluated regarding each experimental diet. At 14, 21, 35 and 42 days of age, the feed intake, weight gain, feed conversion and viability of the chickens were evaluated. At 42 days, the live weight, carcass yield, prime cuts and relative weight of the edible offal were measured. The dietary inclusion of either millet or corn oil did not affect any of the parameters. In conclusion, additions of millet and corn oil to sorghum-based diets of broilers do not compromise poultry performance.

  12. Inclusion of sweet sorghum flour in bread formulations | Araujo ...

    African Journals Online (AJOL)

    Sweet sorghum (Sorghum bicolor L. Moench) has been studied as an additional source of raw material for production or partial replacement of foods due to its high fiber concentration. Its consumption is associated with the prevention of some diseases and nutritional benefits. The aim of this study was to evaluate the partial ...

  13. Phenolic Compositions and Antioxidant Activities Differ Significantly among Sorghum Grains with Different Applications

    Directory of Open Access Journals (Sweden)

    Shuyu Shen

    2018-05-01

    Full Text Available Sorghum grains with different applications had different phenolic profiles, which were corresponded to various antioxidant capacities. In this study, total phenolic, proanthocyanidins and flavonoids contents, as well as contents of individual phenolic compounds from sorghum grains with various applications were determined, and their antioxidant capacities were evaluated. Total phenolic contents (TPC and total proanthocyanidins contents (TPAC showed strong correlation with antioxidant activities (r > 0.95, p < 0.01. Hongyingzi (S-1, one of the brewing sorghums, showed the highest level of TPC and TPAC, while white grain sorghum (S-8 had the lowest. Except for black grain sorghum (S-7, that contained the highest contents of ferulic acid, brewing sorghum grains contained the higher contents of the most individual phenolic compounds, especially the variety S-1. The correlation among individual phenolic compounds and antioxidant activities indicated that the free forms of protocatechuic acid (r = 0.982 of FRAPassay, p < 0.01 and taxifolin (r = 0.826 of FRAP assay, p < 0.01 may be the main functional compounds. These results indicate that brewing sorghum grains can also be utilized as effective materials for functional foods.

  14. Fermentation of sweet sorghum syrup to butanol in the presence of natural nutrients and inhibitors

    Science.gov (United States)

    Sweet sorghum syrups represent a renewable raw material that can be available year-round for production of biofuels and biochemicals. Sweet sorghum sugars have been used as sources for butanol production in the past but most often the studies focused on sweet sorghum juice and not on sweet sorghum s...

  15. An Aspergillus oryzae acetyl xylan esterase: molecular cloning and characteristics of recombinant enzyme expressed in Pichia pastoris.

    Science.gov (United States)

    Koseki, Takuya; Miwa, Yozo; Akao, Takeshi; Akita, Osamu; Hashizume, Katsumi

    2006-02-10

    We screened 20,000 clones of an expressed sequence tag (EST) library from Aspergillus oryzae (http://www.nrib.go.jp/ken/EST/db/index.html) and obtained one cDNA clone encoding a protein with similarity to fungal acetyl xylan esterase. We also cloned the corresponding gene, designated as Aoaxe, from the genomic DNA. The deduced amino acid sequence consisted of a putative signal peptide of 31-amino acids and a mature protein of 276-amino acids. We engineered Aoaxe for heterologous expression in P. pastoris. Recombinant AoAXE (rAoAXE) was secreted by the aid of fused alpha-factor secretion signal peptide and accumulated as an active enzyme in the culture medium to a final level of 190 mg/l after 5 days. Purified rAoAXEA before and after treatment with endoglycosidase H migrated by SDS-PAGE with a molecular mass of 31 and 30 kDa, respectively. Purified rAoAXE displayed the greatest hydrolytic activity toward alpha-naphthylacetate (C2), lower activity toward alpha-naphthylpropionate (C3) and no detectable activity toward acyl-chain substrates containing four or more carbon atoms. The recombinant enzyme catalyzed the release of acetic acid from birchwood xylan. No activity was detectable using methyl esters of ferulic, caffeic or sinapic acids. rAoAXE was thermolabile in comparison to other AXEs from Aspergillus.

  16. cDNA cloning of chicken orexin receptor and tissue distribution: sexually dimorphic expression in chicken gonads.

    Science.gov (United States)

    Ohkubo, T; Tsukada, A; Shamoto, K

    2003-12-01

    Orexin-A and -B are known to stimulate food intake in mammals. However, the critical roles of orexins in birds are not fully understood, since orexins have no stimulatory effect on food intake in the chicken. To understand the physiological role(s) of orexins in birds, we have cloned chicken orexin receptor (cOXR) cDNA by RT-PCR, and analysed the tIssue distribution of OXR mRNA in the chicken. The cOXR cDNA is 1869 bp long and encodes 501 amino acids. The cloned cDNA for cOXR corresponds to the type 2 OXR in mammals, and shows approximately 80% similarity to those of mammals at the amino acid level. Expression analysis by RNase protection assay revealed OXR mRNA was distributed widely in brain regions, and expression in the cerebrum, hypothalamus and optic tectum were abundant. In peripheral tIssues, OXR mRNA was expressed in the pituitary gland, adrenal gland and testis, but no mRNA expression was observed in other tIssues examined. Furthermore, we found that the amount of cOXR mRNA was different between testis and ovary, while prepro-orexin mRNA is equally expressed in the gonads of both sexes in the chicken. These data indicate that the orexins have neuroendocrine actions in chickens, which are mediated through hypothalamic receptors as has been observed in mammals. In addition, orexin may have specific role(s) in the regulation of gonadal function in which sex-dependent mechanisms could be involved.

  17. Hybrid sequencing approach applied to human fecal metagenomic clone libraries revealed clones with potential biotechnological applications.

    Science.gov (United States)

    Džunková, Mária; D'Auria, Giuseppe; Pérez-Villarroya, David; Moya, Andrés

    2012-01-01

    Natural environments represent an incredible source of microbial genetic diversity. Discovery of novel biomolecules involves biotechnological methods that often require the design and implementation of biochemical assays to screen clone libraries. However, when an assay is applied to thousands of clones, one may eventually end up with very few positive clones which, in most of the cases, have to be "domesticated" for downstream characterization and application, and this makes screening both laborious and expensive. The negative clones, which are not considered by the selected assay, may also have biotechnological potential; however, unfortunately they would remain unexplored. Knowledge of the clone sequences provides important clues about potential biotechnological application of the clones in the library; however, the sequencing of clones one-by-one would be very time-consuming and expensive. In this study, we characterized the first metagenomic clone library from the feces of a healthy human volunteer, using a method based on 454 pyrosequencing coupled with a clone-by-clone Sanger end-sequencing. Instead of whole individual clone sequencing, we sequenced 358 clones in a pool. The medium-large insert (7-15 kb) cloning strategy allowed us to assemble these clones correctly, and to assign the clone ends to maintain the link between the position of a living clone in the library and the annotated contig from the 454 assembly. Finally, we found several open reading frames (ORFs) with previously described potential medical application. The proposed approach allows planning ad-hoc biochemical assays for the clones of interest, and the appropriate sub-cloning strategy for gene expression in suitable vectors/hosts.

  18. Cloning and Expression of Cyclophilin from Platanus orientalis Pollens in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Mojtaba Sankian

    2012-10-01

    Full Text Available Background: Allergy is a clinical disorder affecting the human population with wide geographical distribution. Platanus orientalis (P. orientalis trees are planted in many countries and their pollen causes allergic reactions. Cyclophilin has recently been identified as one of the most important allergens of P. orientalis pollen. We aimed to clone and purify this allergen in Escherichia coli for further studies and therapeutic and diagnostic purposes for allergy to P. orientalis. Methods: RNA was extracted from P. orientalis. A full-length fragment encoding cyclophilin was prepared by polymerase chain reaction amplification of the first-strand cDNA synthesized from P. orientalis RNA. The cDNA was inserted into the pET32b (+ vector, and the construct transformed into E. coli Top10 and BL21 cells. The expressed protein was purified by the CuSO4 method. Results: The cDNA for the cyclophilin of P. orientalis pollen was cloned, and a specific reactivity of recombinant cyclophin was confirmed by immunoblotting using sera from patients allergic to P. orientalis pollen. Conclusion: The recombinant cyclophilin has a potential for immunologic assays for evaluation of allergy to P. orientalis pollen.

  19. Sorghum to Ethanol Research Initiative: Cooperative Research and Development Final Report, CRADA Number CRD-08-291

    Energy Technology Data Exchange (ETDEWEB)

    Wolfrum, E.

    2011-10-01

    The goal of this project was to investigate the feasibility of using sorghum to produce ethanol. The work performed included a detailed examination of the agronomics and composition of a large number of sorghum varieties, laboratory experiments to convert sorghum to ethanol, and economic and life-cycle analyses of the sorghum-to-ethanol process. This work showed that sorghum has a very wide range of composition, which depended on the specific sorghum cultivar as well as the growing conditions. The results of laboratory- and pilot-scale experiments indicated that a typical high-biomass sorghum variety performed very similarly to corn stover during the multi-step process required to convert biomass feedstocks to ethanol; yields of ethanol for sorghum were very similar to the corn stover used as a control in these experiments. Based on multi-year agronomic data and theoretical ethanol production, sorghum can achieve more than 1,300 gallons of ethanol per acre given the correct genetics and environment. In summary, sorghum may be a compelling dedicated bioenergy crop that could help provide a portion of the feedstocks required to produce renewable domestic transportation fuels.

  20. A Survey of Viral Diseases of Proso Millet (Panicum miliaceum L. and Sorghum (Sorghum bicolor L. in South Korea

    Directory of Open Access Journals (Sweden)

    Hyun-Geun Min

    2017-09-01

    Full Text Available Throughout year 2015 to 2016, 101 proso millet and 200 sorghum samples were collected from five provinces in South Korea. The samples were subjected to paired-end RNA sequencing and further analyzed by RT-PCR. The results indicated that Rice black-streaked dwarf virus (RBSDV was detected from sorghum collected in Gyeongsang province. The other four viruses, including RBSDV, Rice stripe virus (RSV, Barley virus G (BVG, and Cereal yellow dwarf virus (CYDV, were detected from proso millet. Among four viruses, both RSV and RBSDV were identified high frequency from proso millet collected from Gyeongsang province. Otherwise, BVG was nearly equally identified from five provinces, suggesting that the virus was supposedly widespread nationwide. RBSDV was first identified from both proso millet and sorghum in South Korea. The other virus annotated CYDV identified proso millet was shown to have relatively low identities compared to CYDV previously reported, suggesting that the virus might be new member of Polerovirus.

  1. Cloning and expression of candidate allergens from Culicoides obsoletus for diagnosis of insect bite hypersensitivity in horses

    NARCIS (Netherlands)

    Meide, van der N.M.A.; Roders, N.; Sloet van Oldruitenborgh-Oosterbaan, M.M.; Schaap, P.J.; Oers, van M.M.; Leibold, W.; Savelkoul, H.F.J.; Tijhaar, E.

    2013-01-01

    Insect bite hypersensitivity (IBH) is an IgE-mediated (Type I) hypersensitivity reaction induced by allergens from biting midges of the Culicoides spp. The aim of the present study was to identify, clone and express recombinant allergens from C. obsoletus, the main species found feeding on horses in

  2. Identification of widely varying levels of resistance to meloidogyne incognita in sweet sorghum

    Science.gov (United States)

    Sweet sorghum (Sorghum bicolor) is a potential bioenergy crop that could be incorporated into annual cropping systems in the southern US, where it would likely be rotated with cotton. The desirability of including sweet sorghum in a cotton cropping system will be influenced by sweet sorghum’s host ...

  3. An economic analysis of sweet sorghum cultivation for ethanol production in North China

    NARCIS (Netherlands)

    Liu, H.; Ren, L.; Spiertz, J.H.J.; Zhu, Y.; Xie, G.H.

    2015-01-01

    Sweet sorghum [Sorghum bicolor (L.) Moench] is a promising non-food energy crop. The objective of this study was to determine the economic costs and input sensitivity of sweet sorghum compared to cotton, maize, and sunflower, at two saline-alkali sites in Shandong (Wudi County) and Inner Mongolia

  4. Comparison of sorghum classes for grain and forage yield and forage nutritive value

    Science.gov (United States)

    Sorghum represents a broad category of plants that includes those grown primarily for forage (FS) or grain. Sorghum sudan crosses (SS) are also considered sorghum. Each of these groups can be further classified as brown midrib (BMR), nonBMR, photoperiod sensitive (PS), and nonPS. In our study, sor...

  5. Intake and digestibility of sorghum (Sorghum bicolor, L. Moench silages with different tannin contents in sheep

    Directory of Open Access Journals (Sweden)

    Alex de Matos Teixeira

    2014-01-01

    Full Text Available The purpose of this study was to evaluate the voluntary intake and digestibility of three sorghum (Sorghum bicolor, L. Moench hybrid silages in sheep. The hybrids used were H1 -BRS 655 (CMSXS 222 A × CMSXS 235 R, with tannin; H2 -(ATF54 A × CMSXS 235 R, without tannin; and H3 -BRS 610 (CMSXS 232 A × CMSXS 234 R, without tannin. The intake and digestibility of dry matter (DM, gross energy (GE, neutral detergent fiber (NDF, acid detergent fiber (ADF and crude protein (CP were measured. Eighteen crossbred sheep weighing 59.4 kg (±8.3 were used in the trial. A completely randomized design with three treatments (hybrids and six repetitions (sheep was used. There were no differences in the DM intake or apparent digestibility among the hybrids. Silage of hybrid BRS 610 displayed higher digestibility coefficients for CP, NDF, ADF, and GE compared with the other silages, which did not differ from each other. The neutral detergent fiber, ADF and digestible energy (DE intakes were similar among the hybrids silages. All of the hybrids resulted in a positive N balance in sheep. The levels of DE were superior in hybrid silage BRS 610 in comparison with the other hybrids. Sorghum hybrid BRS 610 silage exhibited superior nutritional value compared with the other hybrids, which is most likely in part due to the absence of tannins. Sorghum silage made with hybrid BRS 610 (CMSXS 232 A × CMSXS 234 R presents superior gross energy, crude protein, neutral detergent fiber and acid detergent fiber digestibility coefficients, as well as greater digestible energy levels than BRS 655 (CMSXS 222 A × CMSXS 235 R and (ATF54 A × CMSXS 235 R.

  6. Direct conversion of sorghum carbohydrates to ethanol by a mixed microbial culture

    Energy Technology Data Exchange (ETDEWEB)

    Christakopoulos, Paul; Lianwu Li; Kekos, Dimitris; Macris, B.J. (National Technical Univ. of Athens (Greece). Dept. of Chemical Engineering)

    1993-01-01

    The carbohydrates of sweet sorghum were directly converted to ethanol by a mixed culture of Fusarium oxysporum F3 and Saccharomyces cerevisiae 2541. A number of factors affecting this bioconversion was studied. Optimum ethanol yields of 33.2 g/100 g of total sorghum carbohydrates, corresponding to 10.3 g/100 g of fresh stalks, were obtained. These values represented 68.6% of the theoretical yield based on total polysaccharides and exceeded that based on oligosaccharides of sorghum by 53.7%. The results demonstrated that more than half of the sorghum polysaccharides were directly fermented to ethanol, thus making the process worthy of further investigation. (author)

  7. Cloning, expression, purification and crystallization of dihydrodipicolinate synthase from the psychrophile Shewanella benthica

    International Nuclear Information System (INIS)

    Wubben, Jacinta M.; Dogovski, Con; Dobson, Renwick C. J.; Codd, Rachel; Gerrard, Juliet A.; Parker, Michael W.; Perugini, Matthew A.

    2010-01-01

    Dihydrodipicolinate synthase (DHDPS) is an essential oligomeric enzyme of interest to antibiotic discovery research and studies probing the importance of quaternary structure to protein function, stability and dynamics. The cloning, expression, purification and crystallization of DHDPS from the psychrophilic (cold-dwelling) bacterium Shewanella benthica are described. Dihydrodipicolinate synthase (DHDPS) is an oligomeric enzyme that catalyzes the first committed step of the lysine-biosynthesis pathway in plants and bacteria, which yields essential building blocks for cell-wall and protein synthesis. DHDPS is therefore of interest to drug-discovery research as well as to studies that probe the importance of quaternary structure to protein function, stability and dynamics. Accordingly, DHDPS from the psychrophilic (cold-dwelling) organism Shewanella benthica (Sb-DHDPS) was cloned, expressed, purified and crystallized. The best crystals of Sb-DHDPS were grown in 200 mM ammonium sulfate, 100 mM bis-tris pH 5.0–6.0, 23–26%(w/v) PEG 3350, 0.02%(w/v) sodium azide and diffracted to beyond 2.5 Å resolution. Processing of diffraction data to 2.5 Å resolution resulted in a unit cell with space group P2 1 2 1 2 1 and dimensions a = 73.1, b = 84.0, c = 143.7 Å. These studies of the first DHDPS enzyme to be characterized from a bacterial psychrophile will provide insight into the molecular evolution of enzyme structure and dynamics

  8. Technical Feasibility and Comprehensive Sustainability Assessment of Sweet Sorghum for Bioethanol Production in China

    Directory of Open Access Journals (Sweden)

    Xiaolin Yang

    2018-03-01

    Full Text Available Under dual pressures of energy and environmental security, sweet sorghum is becoming one of the most promising feedstocks for biofuel production. In the present study, the technical feasibility of sweet sorghum production was assessed in eight agricultural regions in China using the Sweet Sorghum Production Technique Maturity Model. Three top typical agricultural zones were then selected for further sustainability assessment of sweet sorghum production: Northeast China (NEC, Huang-Huai-Hai Basin (HHHB and Ganxin Region (GX. Assessment results demonstrated that NEC exhibited the best sustainable production of sweet sorghum, with a degree of technical maturity value of 0.8066, followed by HHHB and GX, with corresponding values of 0.7531 and 0.6594, respectively. Prospective economic profitability analysis indicated that bioethanol production from sweet sorghum was not feasible using current technologies in China. More efforts are needed to dramatically improve feedstock mechanization logistics while developing new bioethanol productive technology to reduce the total cost. This study provides insight and information to guide further technological development toward profitable industrialization and large-scale sweet sorghum bioethanol production.

  9. Sugarcane Aphid in Sorghum

    Science.gov (United States)

    Evers, Logan

    2018-01-01

    This article is intended for readers in the production agriculture industry who deal with grain sorghum throughout the growing season. This publication will discuss the impacts of the sugarcane aphid in various crops and the ways to manage and identify them as they continue to advance north.

  10. Gene design, cloning and protein-expression methods for high-value targets at the Seattle Structural Genomics Center for Infectious Disease

    International Nuclear Information System (INIS)

    Raymond, Amy; Haffner, Taryn; Ng, Nathan; Lorimer, Don; Staker, Bart; Stewart, Lance

    2011-01-01

    An overview of one salvage strategy for high-value SSGCID targets is given. Any structural genomics endeavor, particularly ambitious ones such as the NIAID-funded Seattle Structural Genomics Center for Infectious Disease (SSGCID) and Center for Structural Genomics of Infectious Disease (CSGID), face technical challenges at all points of the production pipeline. One salvage strategy employed by SSGCID is combined gene engineering and structure-guided construct design to overcome challenges at the levels of protein expression and protein crystallization. Multiple constructs of each target are cloned in parallel using Polymerase Incomplete Primer Extension cloning and small-scale expressions of these are rapidly analyzed by capillary electrophoresis. Using the methods reported here, which have proven particularly useful for high-value targets, otherwise intractable targets can be resolved

  11. Several varieties of sugar sorghum and their possibilities for alcohol production

    Energy Technology Data Exchange (ETDEWEB)

    Bergeret, P W; Fernandez, P W

    1956-01-01

    To study the possibility of using sugar sorghum as a raw material for the production of industrial alcohol, 17 sugar-sorghum varieties from the USA were grown experimentally under field conditions in Uruguay. The best were White African, Honey (Texas) T.S. 21001, and Axtell, which yielded 35,300, 34,200, and 32,450 kg. of stems (1271), 1539, and 14211.100% alcohol)/ha., respectively. The quantity of alcohol/ha obtained from sugar sorghum is almost 3 times that obtained from corn.

  12. Cloning and Expression Analysis of Phenylalanine Ammonia-Lyase Gene in the Mycelium and Fruit Body of the Edible Mushroom Flammulina velutipes

    Science.gov (United States)

    Yun, Yeo Hong; Koo, Ja Sun

    2015-01-01

    Phenylalanine ammonia-lyase (PAL) gene is known to be expressed in plants, and is involved in the differentiation, growth and synthesis of secondary metabolites. However, its expression in fungi remains to be explored. To understand its expression in mushroom fungi, the PAL gene of the edible mushroom Flammulina velutipes (Fvpal) was cloned and characterized. The cloned Fvpal consists of 2,175 bp, coding for a polypeptide containing 724 amino acids and having 11 introns. The translated amino acid sequence of Fvpal shares a high identity (66%) with that of ectomycorrhizal fungus Tricholoma matsutake. Distinctively, the Fvpal expression in the mycelium was higher in minimal medium supplemented with L-tyrosine than with other aromatic amino acids. During cultivation of the mushroom on sawdust medium, Fvpal expression in the fruit body correspondingly increased as the mushroom grew. In the fruiting body, Fvpal was expressed more in the stipe than in the pileus. These results suggest that F. velutipes PAL activity differs in the different organs of the mushroom. Overall, this is first report to show that the PAL gene expression is associated with mushroom growth in fungi. PMID:26539050

  13. Impact of the Soak and the Malt on the Physicochemical Properties of the Sorghum Starches

    Directory of Open Access Journals (Sweden)

    Huiming Zhou

    2010-08-01

    Full Text Available Starches were isolated from soaked and malted sorghum and studied to understand their physicochemical and functional properties. The swelling power (SP and the water solubility index (WSI of both starches were nearly similar at temperatures below 50 °C, but at more than 50 °C, the starch isolated from malted sorghum showed lower SP and high WSI than those isolated from raw and soaked sorghum. The pasting properties of starches determined by rapid visco-analyzer (RVA showed that malted sorghum starch had a lower viscosity peak value (86 BU/RVU than raw sorghum starch (454 BU/RVU. For both sorghum, X-ray diffractograms exhibited an A-type diffraction pattern, typical of cereal starches and the relative degrees of crystallinity ranged from 9.62 to 15.50%. Differential scanning calorimetry (DSC revealed that raw sorghum starch showed an endotherm with a peak temperature (Tp at 78.06 °C and gelatinization enthalpies of 2.83 J/g whereas five-day malted sorghum starch had a Tp at 47.22 °C and gelatinization enthalpies of 2.06 J/g. Storage modulus (G′ and loss modulus (G″ of all starch suspensions increased steeply to a maximum at 70 °C and then decreased with continuous heating. The structural analysis of malted sorghum starch showed porosity on the granule’s surface susceptible to the amylolysis. The results showed that physicochemical and functional properties of sorghum starches are influenced by soaking and malting methods.

  14. Nutritional impact on gene expression and competence of oocytes used to support embryo development and livebirth by cloning procedures in goats.

    Science.gov (United States)

    Fernandes, C C L; Aguiar, L H; Calderón, C E M; Silva, A M; Alves, J P M; Rossetto, R; Bertolini, L R; Bertolini, M; Rondina, D

    2018-01-01

    Changes in the nutritional plan have been shown to affect oocyte quality, crucial to oocyte donors animals used in cloning. This study aimed to evaluate the impact of diets with increasing nutritional levels (maintenance diet=M; 1.3M; 1.6M; 1.9M) fed to goats for four weeks on follicular fluid composition, gene expression and oocyte competence used to cloning in goats. Donor females were superovulated for the retrieval of matured oocytes and physical measurements reported. After four weeks, groups receiving diets above maintenance increased thickness of subcutaneous adipose tissue and body weight, with higher values in 1.9M Group (Pdiet did not affect the expression of GDF9, BMP15, and BAX genes in oocytes, but BCL2 and apoptotic index were significantly higher (P<0.05) in the 1.3M and 1.6M groups than the other groups. Following the transfer of cloned embryos, one fetus was born live of a twin pregnancy in the 1.9M Group. The association between energy intake and oocyte quality suggests better nutritional use by oocytes when the maximum flow was used (1.9M), but the optimal feeding level in cloning still needs refinement. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Molecular Cloning, Expression and Characterization of Pectin Methylesterase (CtPME) from Clostridium thermocellum.

    Science.gov (United States)

    Rajulapati, Vikky; Goyal, Arun

    2017-05-01

    Many phytopathogenic micro-organisms such as bacteria and fungi produce pectin methylesterases (PME) during plant invasion. Plants and insects also produce PME to degrade plant cell wall. In the present study, a thermostable pectin methylesterase (CtPME) from Clostridium thermocellum belonging to family 8 carbohydrate esterase (CE8) was cloned, expressed and purified. The amino acid sequence of CtPME exhibited similarity with pectin methylesterase from Erwinia chrysanthemi with 38% identity. The gene encoding CtPME was cloned into pET28a(+) vector and expressed using Escherichia coli BL21(DE3) cells. The recombinant CtPME expressed as a soluble protein and exhibited a single band of molecular mass approximately 35.2 kDa on SDS-PAGE gels. The molecular mass, 35.5 kDa of the enzyme, was also confirmed by MALDI-TOF MS analysis. Notably, highest protein concentration (11.4 mg/mL) of CtPME was achieved in auto-induction medium, as compared with LB medium (1.5 mg/mL). CtPME showed maximum activity (18.1 U/mg) against citrus pectin with >85% methyl esterification. The optimum pH and temperature for activity of CtPME were 8.5 and 50 °C, respectively. The enzyme was stable in pH range 8.0-9.0 and thermostable between 45 and 70 °C. CtPME activity was increased by 40% by 5 mM Ca 2+ or Mg 2+ ions. Protein melting curve of CtPME gave a peak at 80 °C. The peak was shifted to 85 °C in the presence of 5 mM Ca 2+ ions, and the addition of 5 mM EDTA shifted back the melting peak to 80 °C. CtPME can be potentially used in food and textile industry applications.

  16. Cloning and expression of recombinant, functional ricin B chain

    International Nuclear Information System (INIS)

    Chang, M.S.; Russell, D.W.; Uhr, J.W.; Vitetta, E.S.

    1987-01-01

    The cDNA encoding the B chain of the plant toxin ricin has been cloned and expressed in monkey kidney COS-M6 cells. The recombinant B chain was detected by labeling the transfected cells with [ 35 S]methionine and [ 35 S]-cysteine and demonstrating the secretion of a protein with a M/sub r/ of 30,000-32,000 that was not present in the medium of mock-transfected COS-M6 cells. This protein was specifically immunoprecipitated by an anti-ricin or anti-B-chain antibody and the amount of recombinant B chain secreted by the COS-M6 cells was determined by a radioimmunoassay. Virtually all of the recombinant B chain formed active ricin when mixed with native A chain; it could also bind to the galactose-containing glycoprotein asialofetuin as effectively as native B chain.These results indicate that the vast majority of recombinant B chains secreted into the medium of the COS-M6 cells retain biological function

  17. Hybrid sequencing approach applied to human fecal metagenomic clone libraries revealed clones with potential biotechnological applications.

    Directory of Open Access Journals (Sweden)

    Mária Džunková

    Full Text Available Natural environments represent an incredible source of microbial genetic diversity. Discovery of novel biomolecules involves biotechnological methods that often require the design and implementation of biochemical assays to screen clone libraries. However, when an assay is applied to thousands of clones, one may eventually end up with very few positive clones which, in most of the cases, have to be "domesticated" for downstream characterization and application, and this makes screening both laborious and expensive. The negative clones, which are not considered by the selected assay, may also have biotechnological potential; however, unfortunately they would remain unexplored. Knowledge of the clone sequences provides important clues about potential biotechnological application of the clones in the library; however, the sequencing of clones one-by-one would be very time-consuming and expensive. In this study, we characterized the first metagenomic clone library from the feces of a healthy human volunteer, using a method based on 454 pyrosequencing coupled with a clone-by-clone Sanger end-sequencing. Instead of whole individual clone sequencing, we sequenced 358 clones in a pool. The medium-large insert (7-15 kb cloning strategy allowed us to assemble these clones correctly, and to assign the clone ends to maintain the link between the position of a living clone in the library and the annotated contig from the 454 assembly. Finally, we found several open reading frames (ORFs with previously described potential medical application. The proposed approach allows planning ad-hoc biochemical assays for the clones of interest, and the appropriate sub-cloning strategy for gene expression in suitable vectors/hosts.

  18. Molecular cloning and expression analysis of annexin A2 gene in sika deer antler tip

    Directory of Open Access Journals (Sweden)

    Yanling Xia

    2018-04-01

    Full Text Available Objective Molecular cloning and bioinformatics analysis of annexin A2 (ANXA2 gene in sika deer antler tip were conducted. The role of ANXA2 gene in the growth and development of the antler were analyzed initially. Methods The reverse transcriptase polymerase chain reaction (RT-PCR was used to clone the cDNA sequence of the ANXA2 gene from antler tip of sika deer (Cervus Nippon hortulorum and the bioinformatics methods were applied to analyze the amino acid sequence of Anxa2 protein. The mRNA expression levels of the ANXA2 gene in different growth stages were examined by real time reverse transcriptase polymerase chain reaction (real time RT-PCR. Results The nucleotide sequence analysis revealed an open reading frame of 1,020 bp encoding 339 amino acids long protein of calculated molecular weight 38.6 kDa and isoelectric point 6.09. Homologous sequence alignment and phylogenetic analysis indicated that the Anxa2 mature protein of sika deer had the closest genetic distance with Cervus elaphus and Bos mutus. Real time RT-PCR results showed that the gene had differential expression levels in different growth stages, and the expression level of the ANXA2 gene was the highest at metaphase (rapid growing period. Conclusion ANXA2 gene may promote the cell proliferation, and the finding suggested Anxa2 as an important candidate for regulating the growth and development of deer antler.

  19. Cloning and baculovirus expression of a desiccation stress gene from the beetle, Tenebrio molitor.

    Science.gov (United States)

    Graham, L A; Bendena, W G; Walker, V K

    1996-02-01

    The cDNA sequence encoding a novel desiccation stress protein (dsp28) found in the hemolymph of the common yellow mealworm beetle, Tenebrio molitor, has been determined. The sequence encodes a 225 amino acid protein containing a 20 amino acid signal peptide. Dsp28 shows no significant similarity to any known nucleic acid or protein sequence. Levels of dsp28 mRNA were found to increase approx 5-fold following desiccation. Dsp28 cDNA has been cloned into a baculovirus expression vector and the expressed protein was compared to native dsp28. Both dsp28 expressed by recombinant baculovirus and native dsp28 are glycosylated and N-terminally processed. Although dsp28 is induced by cold in addition to desiccation stress, it does not contribute to the freezing point depression (thermal hysteresis) observed in Tenebrio hemolymph.

  20. Global Expression in Sorghum Brown Midrib (bmr) 6 and 12 Mutants; a Tool to Improve Biomass for Biofuels

    Science.gov (United States)

    Brown midrib (bmr) mutants are being investigated for their ability to increase the conversion efficiency of sorghum biomass for lignocellulosic bioenergy. Brown midrib 6 and 12 (bmr6 and 12) are impaired in the last two steps of monolignol biosynthesis resulting in reduced lignin content and alter...

  1. The Effect of Silicon on some Morpho-physiological Characteristics and Grain Yield of Sorghum (Sorghum bicolor L.) under Salt Stress

    OpenAIRE

    S Hasibi; H Farahbakhsh; Gh Khajoeinejad

    2016-01-01

    Introduction Nowadays, salinity is one of the limiting factors for crop production in arid and semi-arid regions. On the other hand, sorghum (Sorghum bicolor L.) is a self-pollinated and short-day plant, which partly has been adapted to salinity and water stress conditions; also play an important role in humans, livestock and poultry nourishments. All studies have showed the positive effects of Silicon on growth and yield of plants in both normal and stress conditions. The aim of this exp...

  2. CD4+ T-cell clones obtained from cattle chronically infected with Fasciola hepatica and specific for adult worm antigen express both unrestricted and Th2 cytokine profiles.

    Science.gov (United States)

    Brown, W C; Davis, W C; Dobbelaere, D A; Rice-Ficht, A C

    1994-01-01

    The well-established importance of helper T (Th)-cell subsets in immunity and immunoregulation of many experimental helminth infections prompted a detailed study of the cellular immune response against Fasciola hepatica in the natural bovine host. T-cell lines established from two cattle infected with F. hepatica were characterized for the expression of T-cell surface markers and proliferative responses against F. hepatica adult worm antigen. Parasite-specific T-cell lines contained a mixture of CD4+, CD8+, and gamma/delta T-cell-receptor-bearing T cells. However, cell lines containing either fewer than 10% CD8+ T cells or depleted of gamma/delta T cells proliferated vigorously against F. hepatica antigen, indicating that these T-cell subsets are not required for proliferative responses in vitro. Seventeen F. hepatica-specific CD4+ Th-cell clones were examined for cytokine expression following concanavalin A stimulation. Biological assays to measure interleukin-2 (IL-2) or IL-4, gamma interferon (IFN-gamma), and tumor necrosis factor and Northern (RNA) blot analysis to verify the expression of IL-2, IL-4, and IFN-gamma revealed that the Th-cell clones expressed a spectrum of cytokine profiles. Several Th-cell clones were identified as Th2 cells by the strong expression of IL-4 but little or no IL-2 or IFN-gamma mRNA. The majority of Th-cell clones were classified as Th0 cells by the expression of either all three cytokines or combinations of IL-2 and IL-4 or IL-4 and IFN-gamma. No Th1-cell clones were obtained. All of the Th-cell clones expressed a typical memory cell surface phenotype, characterized as CD45Rlow, and all expressed the lymph node homing receptor (L selectin). These results are the first to describe cytokine responses of F. hepatica-specific T cells obtained from infected cattle and extend our previous analysis of Th0 and Th1 cells from cattle immune to Babesia bovis (W. C. Brown, V. M. Woods, D. A. E. Dobbelaere, and K. S. Logan, Infect. Immun. 61

  3. Cloning and expression of pab gene of M. tuberculosis isolated from pulmonary TB patient in E.coli DH5α

    Directory of Open Access Journals (Sweden)

    Tri Y. M. Raras

    2011-11-01

    Full Text Available Background: Mycobacterium tuberculosis antigen38 is a potent serodiagnostic agent containing two M. tuberculosisspecific B-cell epitopes. The high price of imported diagnostic agents hinders realization of fast clinical TB diagnosis in developing countries. Therefore, we produced recombinant antigen38 (recAg38M from M. tuberculosis local strain, which might be used to produce economical tuberculosis serodiagnostic kit.Methods: Pab gene that was isolated from pulmonary TB patient in Malang was cloned into a plasmid vector (pGEMTeasy to construct pMB38. The E.coli DH5α clone carrying pMb38 was selected on X-gal medium. The expression of pab was mediated using pPRoExHTc under the control of Trc promoter and E.coli DH5α as host.Results: Alignment of the pab sequence from the white E.coli DH5α clones with that of M. tuberculosis H37Rv showed 98% homology. The recombinant protein in which the signal peptide has been deleted to prevent the protein being secreted into medium was found in the cytoplasm.Conclusion: pab gene of M. tuberculosis isolated from a TB patient could be expressed in heterologous system in E.coliDH5α. (Med J Indones 2011; 20:247-54Keywords: Mycobacterium tuberculosis, Pab gene expression, recombinant antigen38

  4. FEEDING BROWN MIDRIB FORAGE SORGHUM SILAGE AND CORN GLUTEN FEED TO LACTATING DAIRY COWS

    Science.gov (United States)

    Brown midrib (BMR) forage sorghum contains less lignin , resulting in increased NDF digestibility compared to conventional sorghum . An experiment was conducted to evaluate the effects of BMR forage sorghum silage in diets containing wet corn gluten feed (WCGF). The objective was to determine the e...

  5. Influence of Chemical Treatments Sequence on Morphology and Crystallinity of Sorghum Fibers

    Directory of Open Access Journals (Sweden)

    Ismojo Ismojo

    2018-05-01

    Full Text Available Micro-fibrillated cellulose (MFC derived from natural fibre is continuously gaining interest to produce an environmentally-friendly material, due to economic and ecological reasons. In consequence, sorghum is one of the most-cultivated crops that usually remain the waste as by product of bioethanol production. Indeed, it will be a promising area to utilize sorghum waste to produce MFC for enhancing polymer performance, especially in terms of crystallinity. The objective of this study is to investigate the effect of a sequence of chemical modification was applied to sorghum fibres, i.e. alkalization using 4% sodium hydroxide followed by bleaching using 1.7% sodium chlorite plus acetic acid as a buffer. The treatment was purposed to unbundle the lignocellulose networks into microfibrils cellulose with less amorphous part and lower hydrophilic properties. Evaluation of the chemical treatments effect on internal microstructure, crystallinity index and chemical composition of sorghum fibre was measured via Field-Emission Scanning Electron microscope (FE-SEM, X-ray Diffraction (XRD and Fourier Transformation Infra-Red (FTIR Spectroscopy. The experiments show that treatments led to a removal of binding materials, such as amorphous parts hemicellulose and lignin, from the sorghum fibres, resulting MFC of sorghum fibres and enhanced crystallinity index from 41.12 % to 75.73%.

  6. Production of rhesus monkey cloned embryos expressing monomeric red fluorescent protein by interspecies somatic cell nuclear transfer

    International Nuclear Information System (INIS)

    Zhu, Hai-Ying; Kang, Jin-Dan; Li, Suo; Jin, Jun-Xue; Hong, Yu; Jin, Long; Guo, Qing; Gao, Qing-Shan; Yan, Chang-Guo; Yin, Xi-Jun

    2014-01-01

    Highlights: • Rhesus monkey cells were electroporated with a plasmid containing mRFP1, and an mRFP1-expressing cell line was generated. • For the first time, mRFP1-expressing rhesus monkey cells were used as donor cells for iSCNT. • The effect of VPA on the development of embryos cloned using iSCNT was determined. - Abstract: Interspecies somatic cell nuclear transfer (iSCNT) is a promising method to clone endangered animals from which oocytes are difficult to obtain. Monomeric red fluorescent protein 1 (mRFP1) is an excellent selection marker for transgenically modified cloned embryos during somatic cell nuclear transfer (SCNT). In this study, mRFP-expressing rhesus monkey cells or porcine cells were transferred into enucleated porcine oocytes to generate iSCNT and SCNT embryos, respectively. The development of these embryos was studied in vitro. The percentage of embryos that underwent cleavage did not significantly differ between iSCNT and SCNT embryos (P > 0.05; 71.53% vs. 80.30%). However, significantly fewer iSCNT embryos than SCNT embryos reached the blastocyst stage (2.04% vs. 10.19%, P < 0.05). Valproic acid was used in an attempt to increase the percentage of iSCNT embryos that developed to the blastocyst stage. However, the percentages of embryos that underwent cleavage and reached the blastocyst stage were similar between untreated iSCNT embryos and iSCNT embryos treated with 2 mM valproic acid for 24 h (72.12% vs. 70.83% and 2.67% vs. 2.35%, respectively). These data suggest that porcine-rhesus monkey interspecies embryos can be generated that efficiently express mRFP1. However, a significantly lower proportion of iSCNT embryos than SCNT embryos reach the blastocyst stage. Valproic acid does not increase the percentage of porcine-rhesus monkey iSCNT embryos that reach the blastocyst stage. The mechanisms underling nuclear reprogramming and epigenetic modifications in iSCNT need to be investigated further

  7. Production of rhesus monkey cloned embryos expressing monomeric red fluorescent protein by interspecies somatic cell nuclear transfer

    Energy Technology Data Exchange (ETDEWEB)

    Zhu, Hai-Ying; Kang, Jin-Dan; Li, Suo; Jin, Jun-Xue; Hong, Yu; Jin, Long; Guo, Qing; Gao, Qing-Shan; Yan, Chang-Guo; Yin, Xi-Jun, E-mail: yinxj33@msn.com

    2014-02-21

    Highlights: • Rhesus monkey cells were electroporated with a plasmid containing mRFP1, and an mRFP1-expressing cell line was generated. • For the first time, mRFP1-expressing rhesus monkey cells were used as donor cells for iSCNT. • The effect of VPA on the development of embryos cloned using iSCNT was determined. - Abstract: Interspecies somatic cell nuclear transfer (iSCNT) is a promising method to clone endangered animals from which oocytes are difficult to obtain. Monomeric red fluorescent protein 1 (mRFP1) is an excellent selection marker for transgenically modified cloned embryos during somatic cell nuclear transfer (SCNT). In this study, mRFP-expressing rhesus monkey cells or porcine cells were transferred into enucleated porcine oocytes to generate iSCNT and SCNT embryos, respectively. The development of these embryos was studied in vitro. The percentage of embryos that underwent cleavage did not significantly differ between iSCNT and SCNT embryos (P > 0.05; 71.53% vs. 80.30%). However, significantly fewer iSCNT embryos than SCNT embryos reached the blastocyst stage (2.04% vs. 10.19%, P < 0.05). Valproic acid was used in an attempt to increase the percentage of iSCNT embryos that developed to the blastocyst stage. However, the percentages of embryos that underwent cleavage and reached the blastocyst stage were similar between untreated iSCNT embryos and iSCNT embryos treated with 2 mM valproic acid for 24 h (72.12% vs. 70.83% and 2.67% vs. 2.35%, respectively). These data suggest that porcine-rhesus monkey interspecies embryos can be generated that efficiently express mRFP1. However, a significantly lower proportion of iSCNT embryos than SCNT embryos reach the blastocyst stage. Valproic acid does not increase the percentage of porcine-rhesus monkey iSCNT embryos that reach the blastocyst stage. The mechanisms underling nuclear reprogramming and epigenetic modifications in iSCNT need to be investigated further.

  8. Solid-state fermentation from dried sweet sorghum stalk for bioethanol production

    Energy Technology Data Exchange (ETDEWEB)

    Almodares, A.; Etemadifar, Z.; Omidi, A. [Univ. of Isfahan, Biology Dept., Univ. of Isfahan, Isfahan (Iran, Islamic Republic of)], e-mail: aalmodares@yahoo.com

    2012-11-01

    Due to depletion of global crude oil, countries are interested to alternate fuel energy resources. Presently bioethanol as a source of energy has been a subject of great interest for the industrialized countries. Therefore, there is need for efficient bioethanol production with low cost raw material and production process. Among energy crops, sweet sorghum is the best candidate for bioethanol production. It has been identified as having higher drought tolerance, lower input cost and higher biomass yield than other energy crops. In addition it has wide adoptability and tolerance to abiotic stresses. Moreover due to the shortage of water in dry and hot countries there is a need to reduce water requirement for bioethanol production and solid state fermentation could be the best process for making bioethanol in these countries. The purpose of this study is to achieve the highest ethanol production with lowest amount of water in solid state fermentation using sweet sorghum stalk. In this study the sweet sorghum particles were used for solid state fermentation. Fermentation medium were: sweet sorghum particles with nutrient media, active yeast powder and different moisture contents. The fermentation medium was incubated for 2-3 days at 30 deg C temperature. The results showed sweet sorghum particles (15% w/w) fermented in medium containing 0.5% yeast inoculums, 73.5% moisture content and 3 days incubation period produced the highest amount of ethanol (13% w/w sorghum)

  9. Cloning, expression and purification of d-tagatose 3-epimerase gene from Escherichia coli JM109.

    Science.gov (United States)

    He, Xiaoliang; Zhou, Xiaohui; Yang, Zi; Xu, Le; Yu, Yuxiu; Jia, Lingling; Li, Guoqing

    2015-10-01

    An unknown d-tagatose 3-epimerase (DTE) containing a IoIE domain was identified and cloned from Escherichia coli. This gene was subcloned into the prokaryotic expression vector pET-15b, and induced by IPTG in E. coli BL21 expression system. Through His-select gel column purification and fast-protein liquid chromatography, highly purified and stable DTE protein was produced. The molecular weight of the DTE protein was estimated to be 29.8kDa. The latest 83 DTE sequences from public database were selected and analyzed by molecular clustering, multi-sequence alignment. DTEs were roughly divided into five categories. Copyright © 2015 Elsevier Inc. All rights reserved.

  10. Review of genetic basis of protein digestibility in Grain sorghum

    Science.gov (United States)

    Sorghum, an ancient crop of the semiarid tropics, plays a key role in food and nutritional security for over half-a-billion people in Africa and Asia. In industrialized nations, sorghum is cultivated as animal feed and more recently as a feedstock for biofuel production and as health food alternativ...

  11. Sorghum cobalt analysis on not determined wave length with atomic ...

    African Journals Online (AJOL)

    This study was to know the better wave length on measuring cobalt content in forage sorghum hybrid (Sorghum bicolor) with an atomic absorption spectrophotometer. The analysis was on background correction mode with three wave lengths; 240.8, 240.7 (determined wave length or recommended wave length) and 240.6 ...

  12. Omission and Resupply of Nitrogen Affect Physiological and Enzymatic Activities and the Gene Expression of Eucalypt Clones

    Directory of Open Access Journals (Sweden)

    Loane Vaz Fernandes

    Full Text Available ABSTRACT: The mineral nutrient uptake of plants in the field occurs in pulses, due to variations in the substance concentrations at the root surface. The fluctuations in nutrient supply probably induce changes in the plant, which are to date unknown for Eucalyptus. This study evaluated these changes in plant growth, nutritional status, photosynthesis, and gene expression, which can serve as biomarkers of the nitrogen status, of four eucalypt clones exposed to N omission and resupply. A greenhouse experiment with four Eucalyptus clones was installed, and after initial growth exposed to N omission for 21 d, followed by N resupply in nutrient solution for 14 d. Nitrogen omission decreased the total N and photosynthetic pigments, net photosynthesis and photochemical dissipation, and increased enzyme activity especially in leaves and the gene expression in leaves and roots. Nitrogen resupply decreased these variations, indicating recovery. The total N concentration was highly and significantly correlated with net photosynthesis, enzyme activity, expression of genes GS2;1 and Gln1;3 in the leaves and AMT1;2 in the roots, contents of chlorophyll a and b, and photochemical energy dissipation. The enzymes GS and NR in the leaves and the genes AMT1;2, GS2;1 and Gln1;3 proved to be sensitive N indicators.

  13. Molecular cloning of cDNA for rat brain metallothionein-2 and regulation of its gene expression

    Energy Technology Data Exchange (ETDEWEB)

    Saijoh, Kiyofumi; Sumino, Kimiaki [Department of Public Health, Kobe University School of Medicine (Japan); Kuno, Takayoshi; Shuntoh, Hisato; Tanaka, Chikako [Department of Pharmacology, Kobe University of Medicine (Japan)

    1989-01-01

    A rat brain metallothionein-II (MT-II) complementary DNA (cDNA) clone was isolated from a cDNA plasmid library, which was prepared from non-treated rat brain mRNA, by a colony screening procedure using /sup 32/P-labeled synthetic oligonucleotide probes. It is deduced that the clone encodes for a protein of 61 amino acids comprising 20 cysteines, which is highly homologous to MT-IIs in other species. Northern blot analysis demonstrated major mRNA species in the brain, liver and kidneys (approximately 350 b in size), which is induced in response to dexamethasone, zinc, cadmium and mercury but not to methyl mercury. These findings confirm that MT-II genes are expressed and regulated both by steroid and heavy metals in the brain as well as in peripheral organs. (author).

  14. Molecular cloning of cDNA for rat brain metallothionein-2 and regulation of its gene expression

    International Nuclear Information System (INIS)

    Saijoh, Kiyofumi; Sumino, Kimiaki; Kuno, Takayoshi; Shuntoh, Hisato; Tanaka, Chikako

    1989-01-01

    A rat brain metallothionein-II (MT-II) complementary DNA (cDNA) clone was isolated from a cDNA plasmid library, which was prepared from non-treated rat brain mRNA, by a colony screening procedure using 32 P-labeled synthetic oligonucleotide probes. It is deduced that the clone encodes for a protein of 61 amino acids comprising 20 cysteines, which is highly homologous to MT-IIs in other species. Northern blot analysis demonstrated major mRNA species in the brain, liver and kidneys (approximately 350 b in size), which is induced in response to dexamethasone, zinc, cadmium and mercury but not to methyl mercury. These findings confirm that MT-II genes are expressed and regulated both by steroid and heavy metals in the brain as well as in peripheral organs. (author)

  15. Cloning and shake flask expression of hrIDS- Like in Pichia pastoris ...

    African Journals Online (AJOL)

    The human Iduronate-2-sulfate sulfatase (hIDS-Like) was cloned into the methylotrophic yeast Pichia pastoris under the control of alcohol oxidase promoter (AOX1) and the -mating factor signal peptide (a-factor). Six clones were identified by PCR. Using clone IDS28, the enzyme was secreted into the culture medium, ...

  16. Cloning of zebrafish Mustn1 orthologs and their expression during early development.

    Science.gov (United States)

    Camarata, Troy; Vasilyev, Aleksandr; Hadjiargyrou, Michael

    2016-11-15

    Mustn1 is a small nuclear protein that is involved in the development and regeneration of the musculoskeletal system. Previous work established a role for Mustn1 in myogenic and chondrogenic differentiation. In addition, recent evidence suggests a potential role for Mustn1 in cilia function in zebrafish. A detailed study of Mustn1 expression has yet to be conducted in zebrafish. As such, we report herein the cloning of the zebrafish Mustn1 orthologs, mustn1a and mustn1b, and their expression during zebrafish embryonic and larval development. Results indicate a 44% nucleotide identity between the two paralogs. Phylogenetic analysis further confirmed that the Mustn1a and 1b predicted proteins were highly related to other vertebrate members of the Mustn1 protein family. Whole mount in situ hybridization revealed expression of both mustn1a and 1b at the 7-somite stage through 72hpf in structures such as Kupffer's vesicle, segmental mesoderm, head structures, and otic vesicle. Additionally, in 5day old larva, mustn1a and 1b expression is detected in the neurocranium, otic capsule, and the gut. Although both were expressed in the neurocranium, mustn1a was localized in the hypophyseal fenestra whereas mustn1b was found near the posterior basicapsular commissure. mustn1b also displayed expression in the ceratohyal and ceratobranchial elements of the pharyngeal skeleton. These expression patterns were verified temporally by q-PCR analysis. Taken together, we conclude that Mustn1 expression is conserved in vertebrates and that the variations in expression of the two zebrafish paralogs suggest different modes of molecular regulation. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. EasyClone-MarkerFree

    DEFF Research Database (Denmark)

    Fabre, Mathew Malcolm Jessop; Jakociunas, Tadas; Stovicek, Vratislav

    2016-01-01

    Clone-MarkerFree. The integration of linearized expression cassettes into defined genomic loci is facilitated by CRISPR/Cas9. Cas9 is recruited to the chromosomal location by specific guide RNAs (gRNAs) expressed from a set of gRNA helper vectors. Using our genome engineering vector suite, single and triple insertions are obtained...

  18. Analysis of sorghum wax and carnauba wax by reversed phase liquid chromatography mass spectrometry

    Science.gov (United States)

    Sorghum is a genus in the grass family, which is used for both grain and forage production throughout the world. In the United States, sorghum grain is predominantly used as livestock feed, and in ethanol production. In recent years however, sorghum grain has been investigated for other industrial a...

  19. Dicty_cDB: SSL552 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 28 0.36 3 BZ330174 |BZ330174.1 hv90e08.b1 WGS-SbicolorF (JM107 adapted methyl filtered) Sorghum bicolor gen...S-SbicolorF (JM107 adapted methyl filtered) Sorghum bicolor genomic clone hv90e08 5', DNA sequence. 36 0.63 ...2 BZ335743 |BZ335743.1 hz26f02.g1 WGS-SbicolorF (JM107 adapted methyl filtered) Sorghum bicolor genomic clon...e hz26f02 5', DNA sequence. 36 0.68 2 BZ628823 |BZ628823.1 ih62d12.g1 WGS-SbicolorF (DH5a methyl filtered) S...06.1 ic39h07.b1 WGS-SbicolorF (JM107 adapted methyl filtered) Sorghum bicolor genomic clone ic39h07 5', DNA

  20. Potential of multiseeded mutant (msd) to boost sorghum grain yield

    Science.gov (United States)

    Seed number per plant is an important determinant of the grain yield in cereal and other crops. We have isolated a class of multiseeded (msd) sorghum (Sorghum bicolor L. Moench) mutants that are capable of producing three times the seed number and twice the seed weight per panicle as compared with t...

  1. Evaluation of whorl damage by fall armyworm (Lepidoptera:Noctuidae) on field and greenhouse grown sweet sorghum plants

    Science.gov (United States)

    The fall armyworm [Spodoptera frugiperda (Lepidoptera: Noctuidae)] is an economically important pest of sorghum [Sorghum bicolor (L) Moench]. However, resistance to fall armyworm in sweet sorghum has not been extensively studied. A collection of primarily sweet sorghum accessions were evaluated in t...

  2. Sweet Sorghum Alternative Fuel and Feed Pilot Project

    Energy Technology Data Exchange (ETDEWEB)

    Slack, Donald C. [Univ. of Arizona, Tucson, AZ (United States). Agricultural and Biosystems Engineering Dept.; Kaltenbach, C. Colin [Univ. of Arizona, Tucson, AZ (United States)

    2013-07-30

    The University of Arizona undertook a “pilot” project to grow sweet sorghum on a field scale (rather than a plot scale), produce juice from the sweet sorghum, deliver the juice to a bio-refinery and process it to fuel-grade ethanol. We also evaluated the bagasse for suitability as a livestock feed and as a fuel. In addition to these objectives we evaluated methods of juice preservation, ligno-cellulosic conversion of the bagasse to fermentable sugars and alternative methods of juice extraction.

  3. Sorghum bicolor L. Moench

    African Journals Online (AJOL)

    sorghum plants mitigates the negative effect of drought stress, favoring this crop cultivation in areas of low water ... It is a salt and aluminum-tolerant crop, making areas suitable for ... its growth or decrease its metabolic activity and later, when water ..... and osmoregulation, but also in stabilizing the structures and enzyme ...

  4. Human placental Na+, K+-ATPase α subunit: cDNA cloning, tissue expression, DNA polymorphism, and chromosomal localization

    International Nuclear Information System (INIS)

    Chehab, F.F.; Kan, Y.W.; Law, M.L.; Hartz, J.; Kao, F.T.; Blostein, R.

    1987-01-01

    A 2.2-kilobase clone comprising a major portion of the coding sequence of the Na + , K + -ATPase α subunit was cloned from human placenta and its sequence was identical to that encoding the α subunit of human kidney and HeLa cells. Transfer blot analysis of the mRNA products of the Na + , K + -ATPase gene from various human tissues and cell lines revealed only one band (≅ 4.7 kilobases) under low and high stringency washing conditions. The levels of expression in the tissues were intestine > placenta > liver > pancreas, and in the cell lines the levels were human erythroleukemia > butyrate-induced colon > colon > brain > HeLa cells. mRNA was undetectable in reticulocytes, consistent with the authors failure to detect positive clones in a size-selected ( > 2 kilobases) λgt11 reticulocyte cDNA library. DNA analysis revealed by a polymorphic EcoRI band and chromosome localization by flow sorting and in situ hybridization showed that the α subunit is on the short is on the short arm (band p11-p13) of chromosome 1

  5. Supplementary data: Mapping of shoot fly tolerance loci in sorghum ...

    Indian Academy of Sciences (India)

    Supplementary data: Mapping of shoot fly tolerance loci in sorghum using SSR markers. D. B. Apotikar, D. Venkateswarlu, R. B. Ghorade, R. M. Wadaskar, J. V. Patil and P. L. Kulwal. J. Genet. 90, 59–66. Table 1. List of SSR primers for sorghum. Primer code. Forward and reverse. Annealing temperature (°C). Product.

  6. Cloning and expression of NS3 gene of Pakistani isolate type 2 dengue virus

    Directory of Open Access Journals (Sweden)

    Yasmin Farkhanda

    2018-03-01

    Full Text Available Introduction: Dengue is one of the major emerging viral diseases in the world, with dramatic increases in reported cases in the last few decades and annual worldwide occurrence of approximately 390 million infections. It is a highly important mosquito-vectored disease and is a problem in tropical and subtropical areas of the world. The major aim of this study was to clone and express the dengue NS3 gene, in service to its therapeutic importance for the development of stable cell lines.

  7. cDNA Cloning, expression and characterization of an allergenic 60s ribosomal protein of almond (prunus dulcis).

    Science.gov (United States)

    Abolhassani, Mohsen; Roux, Kenneth H

    2009-06-01

    Tree nuts, including almond (prunus dulcis) are a source of food allergens often associated with life-threatening allergic reactions in susceptible individuals. Although the proteins in almonds have been biochemically characterized, relatively little has been reported regarding the identity of the allergens involved in almond sensitivity. The present study was undertaken to identify the allergens of the almond by cDNA library approach. cDNA library of almond seeds was constructed in Uni-Zap XR lamda vector and expressed in E. coli XL-1 blue. Plaques were immunoscreened with pooled sera of allergic patients. The cDNA clone reacting significantly with specific IgE antibodies was selected and subcloned and subsequently expressed in E. coli. The amino acids deducted from PCR product of clone showed homology to 60s acidic ribosomal protein of almond. The expressed protein was 11,450 Dalton without leader sequence. Immunoreactivity of the recombinant 60s ribosomal protein (r60sRP) was evaluated with dot blot analysis using pooled and individual sera of allergic patients. The data showed that r60sRP and almond extract (as positive control) possess the ability to bind the IgE antibodies. The results showed that expressed protein is an almond allergen.Whether this r60sRP represents a major allergen of almond needs to be further studied which requires a large number of sera from the almond atopic patients and also need to determine the IgE-reactive frequencies of each individual allergen.

  8. Double row spacing and drip irrigation as technical options in energy sorghum management

    Directory of Open Access Journals (Sweden)

    Neri Roncucci

    2014-02-01

    Full Text Available The effect of two row spacing configurations and four water supply levels was investigated on sweet and fibre sorghum in Central Italy for two consecutive years. Results highlighted the influence of both irrigation and row spatial configuration on crop productivity. Indeed, several studies have pointed out the positive response of sorghum to irrigation in Mediterranean climate, as in this environment water stress represents one of the main limiting factors on crop productivity. On the other hand, few attempts have been made to explore the role of row spacing on energy sorghum productivity. Results outlined an average increase in sorghum dry biomass yield ranging from +23% to +79% at variable rates of water supply as compared to rainfed control. The positive effect of irrigation was also observed on leaf area index and radiation use efficiency. Moreover, we observed a crop yield increase, from 9% to 20%, under double row spacing compared to the standard planting pattern (i.e. single row spacing. Finally, it was confirmed the efficient use of water by sorghum and the great ability of sorghum to increase its biomass yield in response to increasing volumes of water supplied. Therefore, this work suggests how row spacing configuration and drip irrigation could be feasible technical options to increase sorghum biomass yields in Mediterranean environments. These techniques should be experienced by farmers towards a sustainable intensification of current cropping systems.

  9. Cloning, expression and activation of a truncated 92-kDa gelatinase minienzyme.

    Science.gov (United States)

    Kröger, M; Tschesche, H

    1997-09-01

    The matrix metalloproteinases (MMPs) are a family of highly homologous zinc-endopeptidases that degrade extracellular matrix components. Human 92-kDa gelatinase (MMP-9) represents one of the MMPs that cleaves native collagen type IV. As a basis for structural investigations, the short form (catalytic domain, amino acid residues 113-450) of the 92-kDa gelatinase cDNA was cloned and expressed in E. coli as a minienzyme. By combination of reverse transcription (RT) and polymerase chain reaction (PCR), the truncated 92-kDa gelatinase-cDNA was amplified from the corresponding mRNA derived from ovarian carcinoma cells. The cDNA fragment obtained was cloned in E. coli and sequenced. With the exception of one nucleotide inversion at position 745 (gt-->tg) the cDNA sequence was identical to the nucleotide sequence of the 92-kDa gelatinase as has been previously reported. The protein was expressed in E. coli using the vector pET-12b. The recombinant protein was stored in inclusion bodies and extracted as a 38 kDa species from the inclusion bodies by solubilization in 8 M urea. The product was purified by affinity chromatography and gel filtration. Amino-terminal sequence analysis confirmed the identity with the catalytic domain of 92-kDa gelatinase. The recombinant protein was refolded in the presence of Ca2+ and Zn2+ and yielded an active minienzyme with gelatinolytic activity. It degrades the native substrate collagen type IV and the synthetic substrate Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 x AcOH like the full-length 92-kDa gelatinase. The catalytic activity could be inhibited by the specific MMP inhibitors TIMP-1 and TIMP-2.

  10. SOME CONSIDERATIONS ON THE PROSPECTS OF SORGHUM CROP

    OpenAIRE

    Agatha POPESCU; Reta CONDEI

    2014-01-01

    The paper purpose was to analyze the sorghum statement at world, EU and Romania level in order to establish the main trends in the future of this crop. Sorghum is an important cereal coming on the 5th position after maize, rice, wheat and barley at world level due to its importance in human nutrition, animal feed, in producing bioethanol and green energy, and due to its good impact on environment. It is cultivated on all the continents, in the tropical, subtropical and temperate areas due to ...

  11. Overexpression of Differentially Expressed Genes Identified in Non-pathogenic and Pathogenic Entamoeba histolytica Clones Allow Identification of New Pathogenicity Factors Involved in Amoebic Liver Abscess Formation.

    Directory of Open Access Journals (Sweden)

    Martin Meyer

    2016-08-01

    Full Text Available We here compared pathogenic (p and non-pathogenic (np isolates of Entamoeba histolytica to identify molecules involved in the ability of this parasite to induce amoebic liver abscess (ALA-like lesions in two rodent models for the disease. We performed a comprehensive analysis of 12 clones (A1-A12 derived from a non-pathogenic isolate HM-1:IMSS-A and 12 clones (B1-B12 derived from a pathogenic isolate HM-1:IMSS-B. "Non-pathogenicity" included the induction of small and quickly resolved lesions while "pathogenicity" comprised larger abscess development that overstayed day 7 post infection. All A-clones were designated as non-pathogenic, whereas 4 out of 12 B-clones lost their ability to induce ALAs in gerbils. No correlation between ALA formation and cysteine peptidase (CP activity, haemolytic activity, erythrophagocytosis, motility or cytopathic activity was found. To identify the molecular framework underlying different pathogenic phenotypes, three clones were selected for in-depth transcriptome analyses. Comparison of a non-pathogenic clone A1np with pathogenic clone B2p revealed 76 differentially expressed genes, whereas comparison of a non-pathogenic clone B8np with B2p revealed only 19 differentially expressed genes. Only six genes were found to be similarly regulated in the two non-pathogenic clones A1np and B8np in comparison with the pathogenic clone B2p. Based on these analyses, we chose 20 candidate genes and evaluated their roles in ALA formation using the respective gene-overexpressing transfectants. We conclude that different mechanisms lead to loss of pathogenicity. In total, we identified eight proteins, comprising a metallopeptidase, C2 domain proteins, alcohol dehydrogenases and hypothetical proteins, that affect the pathogenicity of E. histolytica.

  12. Overexpression of Differentially Expressed Genes Identified in Non-pathogenic and Pathogenic Entamoeba histolytica Clones Allow Identification of New Pathogenicity Factors Involved in Amoebic Liver Abscess Formation.

    Science.gov (United States)

    Meyer, Martin; Fehling, Helena; Matthiesen, Jenny; Lorenzen, Stephan; Schuldt, Kathrin; Bernin, Hannah; Zaruba, Mareen; Lender, Corinna; Ernst, Thomas; Ittrich, Harald; Roeder, Thomas; Tannich, Egbert; Lotter, Hannelore; Bruchhaus, Iris

    2016-08-01

    We here compared pathogenic (p) and non-pathogenic (np) isolates of Entamoeba histolytica to identify molecules involved in the ability of this parasite to induce amoebic liver abscess (ALA)-like lesions in two rodent models for the disease. We performed a comprehensive analysis of 12 clones (A1-A12) derived from a non-pathogenic isolate HM-1:IMSS-A and 12 clones (B1-B12) derived from a pathogenic isolate HM-1:IMSS-B. "Non-pathogenicity" included the induction of small and quickly resolved lesions while "pathogenicity" comprised larger abscess development that overstayed day 7 post infection. All A-clones were designated as non-pathogenic, whereas 4 out of 12 B-clones lost their ability to induce ALAs in gerbils. No correlation between ALA formation and cysteine peptidase (CP) activity, haemolytic activity, erythrophagocytosis, motility or cytopathic activity was found. To identify the molecular framework underlying different pathogenic phenotypes, three clones were selected for in-depth transcriptome analyses. Comparison of a non-pathogenic clone A1np with pathogenic clone B2p revealed 76 differentially expressed genes, whereas comparison of a non-pathogenic clone B8np with B2p revealed only 19 differentially expressed genes. Only six genes were found to be similarly regulated in the two non-pathogenic clones A1np and B8np in comparison with the pathogenic clone B2p. Based on these analyses, we chose 20 candidate genes and evaluated their roles in ALA formation using the respective gene-overexpressing transfectants. We conclude that different mechanisms lead to loss of pathogenicity. In total, we identified eight proteins, comprising a metallopeptidase, C2 domain proteins, alcohol dehydrogenases and hypothetical proteins, that affect the pathogenicity of E. histolytica.

  13. Kajian Nilai Energi Metabolis Biji Sorghum Melalui Teknologi Sangrai Pada Ayam Petelur Periode Afkir

    Directory of Open Access Journals (Sweden)

    Hanny Indrat Wahyuni

    2008-04-01

    Full Text Available Evaluation of metabolic energy value of roasted sorghum in culled laying chickens  ABSTRACT. Tannin contained in sorghum can be reduced by using technology processing such as roasting. By using this way, husk of sorghum can be removed leading to decrease of tannin content which is reflected by the value of metabolism energy. The purpose of this experiment is to investigate the effect of roasted sorghum on metabolism energy of culled laying chickens. Measurement of metabolic energy as mathematic is used as comparison. The material used in his experiment was red sorghum, water, and 39 culled laying chickens. Equipment used in this experiment was balance, roasting tool, plastic, force feeding equipment, metabolism cages and bomb calori-meter. This experiment used completely randomized design consisting of 4 treatments and 4 replications (each replication 3 chickens. Treatment consisted of T0 = no roasted sorghum, T1 = roasted for 5 minutes and T2 = roasted for 10 minutes. Data collected were metabolism energy of roasted sorghum both biologically (force feeding and mathematically (proximate analysis at culled laying chickens. All data were statistically calculated, further statistically was conducted by using Duncan and compression of metabolism energy was calculated by using t-Test. The results show that, no statistically effect (p>0, 05 on duration of roasting on metabolism energy of sorghum. Based on t-Test analysis, there was a significantly difference (p<0, 05 between biological metabolism and mathematical metabolism. From this experiment, it can be concluded that 10 minutes of roasting cannot increase of sorghum metabolic energy. The average of biological metabolic was lower (3105, 94 kcal/kg compared to the average of mathematical metabolic energy (3766, 82 kcal/kg.

  14. The Kraft Pulp And Paper Properties of Sweet Sorghum Bagasse (Sorghum bicolor L Moench

    Directory of Open Access Journals (Sweden)

    Widya Fatriasari

    2015-05-01

    Full Text Available This study investigated the potency of sweet sorghum (Sorghum bicolor bagasse as raw material for pulp and paper using kraft pulping. The effects of alkali and sulfidity loading on kraft pulp and paper properties were also investigated. The pulping condition of the kraft pulp consisted of three levels of alkali loading (17, 19 and 22% and sulfidity loading (20, 22 and 24%. The maximum cooking temperature was 170°C for 4 h with a liquid to wood ratio of 10:1. Kraft pulping of this Numbu bagasse produced good pulp indicated by high screen yield and delignification selectivity with a low Kappa number (< 10. The unbleached pulp sheet produced a superior brightness level and a high burst index. The increase of active alkali loading tended to produce a negative effect on the pulp yield, Kappa number and paper sheet properties. Therefore, it is suggested to use a lower active alkaline concentration.

  15. Cloning and expression of pineapple sucrosephosphate synthase ...

    African Journals Online (AJOL)

    A 1132-base pairs (bp) polymerase-chain-reaction product of sucrose-phosphate synthase (SPS) (EC 2.3.1.14) from pineapple (Ananas comosus cv. Comte de paris) fruit was cloned and nominated as Ac- SPS1. The sequence encodes a putative 377 amino acids protein containing two serine conserved features that had ...

  16. Quality and Quantity of Sorghum Hydroponic Fodder from Different Varieties and Harvest Time

    Science.gov (United States)

    Chrisdiana, R.

    2018-02-01

    This experiment was designed to compare different varieties and harvest time of sorghum hydroponic fodder based on nutrient content and biomass production. Experimental design for fodder productivity was completely randomized design with 2 x 3 factorial, i.e., sorghum varieties (KD 4 and Super-1) and time of harvesting the sorghum hydroponic fodder (8,12 and 16 d). Total biomass and DM production, were affected significantly (p<0.05) on harvest time. Total biomass and nutrient content were increased in longer harvest time. The nutrient content were increased with decreasing total value of DM. Super-1 varieties produce larger biomass and nutrient content higher than KD4 (p<0.05). Based on sorghum hidroponic fodder quality and quantity, sorghum hidroponic fodder with Super-1 varieties harvested at 12 d had a good quality of fodder and it can be alternative of technology providing quality forage and land saving with a short time planting period and continous production.

  17. Path analysis of the productive traits in Sorghum species

    Directory of Open Access Journals (Sweden)

    Ikanović Jela

    2011-01-01

    Full Text Available This research studied the phenotypic correlation coefficients between three Sorghum species, namely forage sorghum S. bicolor Moench. (c. NS-Džin, Sudan grass S. sudanense L. (c. Zora and interspecies hybrid S. bicolor x S. sudanense (c. Siloking. The analyses were performed on plant material samples taken from the first cutting, when plants were in the beginning phase of tasseling. The following morphologic traits were studied: plant height, number of leaves per plant, stem leaf weight and mean stem weight. Additionally, their direct and indirect effect on dependent variable green biomass yield was analyzed, for which path coefficients were calculated. This method enables more quality and full insight into relations existing among the studied traits, more precise establishment of cause-effect connections among them, as well as to separate direct from indirect effects of any particular trait on dependent variable, being biomass yield in this case. The analysis of phenotypic coefficients revealed differences in direct and indirect effect of certain traits on dependent variable. Sudan grass had the highest stem (2.281 m and most leaves per plant (7.917. Forage sorghum had the largest leaf weight per plant (49.05 g, while interspecies hybrid had the highest mean stem weight (80.798 g. Variations of these morphologic traits among species were found to be significant and very significant. Morphologic traits - stem height and weight significantly affected sorghum green biomass yield. Leaf number and leaf portion in total biomass were negatively correlated with yield. Cultivars differed significantly regarding morphologic and productive traits. Sudan grass had the lowest green biomass yield, while forage sorghum and interspecies hybrid had significant yield increase.

  18. Effects of Cerium Oxide Nanoparticles on Sorghum Plant Traits

    Science.gov (United States)

    Mu, L.; Chen, Y.; Darnault, C. J. G.; Rauh, B.; Kresovich, S.; Korte, C.

    2015-12-01

    Nanotechnology and nanomaterials are considered as the development of the modern science. However, besides with that wide application, nanoparticles arouse to the side effects on the environment and human health. As the catalyst of ceramics and fuel industry, Cerium (IV) oxide nanoparticles (CeO2 NPs) can be found in the environment following their use and life-cycle. Therefore, it is critical to assess the potential effects that CeO2 NPs found in soils may have on plants. In this study, CeO2 NPs were analyzed for the potential influence on the sorghum [Sorghum bicolor (L.) Moench] (Reg. no. 126) (PI 154844) growth and traits. The objectives of this research were to determine whether CeO2 NPs impact the sorghum germination and growth characteristics. The sorghum was grown in the greenhouse located at Biosystems Research Complex, Clemson University under different CeO2 NPs treatments (0mg; 100mg; 500mg; 1000mg CeO2 NPs/Kg soil) and harvested around each month. At the end of the each growing period, above ground vegetative tissue was air-dried, ground to 2mm particle size and compositional traits estimated using near-infrared spectroscopy. Also, the NPK value of the sorghum tissue was tested by Clemson Agriculture Center. After the first harvest, the result showed that the height of above ground biomass under the nanoparticles stress was higher than that of control group. This difference between the control and the nanoparticles treatments was significant (F>F0.05; LSD). Our results also indicated that some of the compositional traits were impacted by the different treatments, including the presence and/or concentrations of the nanoparticles.

  19. Productivity and Nutrient Quality of Some Sorghum Mutant Lines at Different Cutting Ages

    Directory of Open Access Journals (Sweden)

    R. E. Puteri

    2015-08-01

    Full Text Available The objective of the study was to explore the appropriate cutting age to produce optimal biomass and good nutrient quality from sorghum mutant lines BMR i.e., PATIR 3.5 M7, PATIR 3.6 M7, and PATIR 3.7 M7, also SAMURAI I (M17. A completely randomized in Split Plot design with 2 factors and 3 replicates was used. The first factor was the type of sorghum (SAMURAI I M17, PATIR 3.5, PATIR 3.6, PATIR 3.7 as the main plot and the second factor was the cutting age (85, 95, 105 as a subplot. Parameters observed were the production of stems, leaves, grains, total biomass production, ash, crude fat, crude fiber, crude protein, NFE, TDN, percentage of DMD, OMD and N-NH3. Data were analyzed by using ANOVA followed by DMRT (Duncan Multiple Range Test. The results showed that there were highly significant interactions (P<0.01 between cutting age and type of sorghum in production of stems, leaves, grains, total biomass production, value of TDN, DMD, OMD, and N-NH3. Increasing cutting age significantly increased the percentage of ash content, crude protein and crude fat. The sorghum type significantly affected crude fat content nonBMR sorghum variety of SAMURAI I (M17 and achieved optimal biomass production and nutrient content at cutting age of 85 d similar to BMR sorghum mutant lines PATIR 3.6 and PATIR 3.5, whereas BMR sorghum mutant lines of PATIR 3.7 achieved optimum production at the age of 95 d of cutting. All types of sorghum varieties was not recommended to be harvested at 105 d. Biomass production increased with the increasing of cutting age, but the nutrient content decreased.

  20. Molecular cloning and gene expression of Foxl2 in the Nile tilapia, Oreochromis niloticus

    International Nuclear Information System (INIS)

    Wang Deshou; Kobayashi, Tohru; Zhou Linyan; Nagahama, Yoshitaka

    2004-01-01

    A Foxl2 cDNA was cloned from the Nile tilapia ovary by RT-PCR and subsequent RACE. Alignment of known Foxl2 sequences from vertebrates confirmed the conservation of the Foxl2 open reading frame and protein sequences, especially the forkhead domain and C-terminal region, while some homopolymeric runs of amino acids are found only in mammals but not in non-mammalian vertebrates. RT-PCR revealed that Foxl2 is expressed in the tilapia brain (B), pituitary (P), gill, and gonads (G), with the highest level of expression in the ovary, reflecting the involvement of Foxl2 in B-P-G axis. Northern blotting and in situ hybridization also revealed an evident sexual dimorphic expression pattern in the gonads. Foxl2 mRNA was mainly detected in the granulosa cells surrounding the oocytes. The ovarian expression of Foxl2 in tilapia begins early during the differentiation of the gonads and persists until adulthood, implying the involvement of Foxl2 in fish gonad differentiation and the maintenance of ovarian function

  1. Pretreatment of sweet sorghum bagasse for hydrogen production by Caldicellulosiruptor saccharolyticus

    NARCIS (Netherlands)

    Panagiotopoulos, I.A.; Bakker, R.R.; Vrije, de G.J.; Koukios, E.G.; Claassen, P.A.M.

    2010-01-01

    Pretreatment of sweet sorghum bagasse, an energy crop residue, with NaOH for the production of fermentable substrates, was investigated. Optimal conditions for the alkaline pretreatment of sweet sorghum bagasse were realized at 10% NaOH (w/w dry matter). A delignification of 46% was then observed,

  2. The effect of alpha amylase enzyme on quality of sweet sorghum juice for chrystal sugar

    Science.gov (United States)

    Marwati, T.; Cahyaningrum, N.; Widodo, S.; Astiati, U. T.; Budiyanto, A.; Wahyudiono; Arif, A. B.; Richana, N.

    2018-01-01

    Sweet sorghum juice (Sorghum bicolor L. Moench) has characteristics similar to sugar cane juice and potentially used for sugar substitutes that can support food security. Nevertheless the sweet sorghum juicecontain starch which impede sorghum sugar crystallization. Therefore, research on the enzymatic process is needed to convert starch into reducing sugar. The experimental design used was the Factorial Randomized Design with the first factor was alpha amylase enzyme concentration (0, 20, 40, 60, 80, 100, 120 μL/100 mL) and second factor was incubation time (0, 30, 60, 90 minute) at temperature 100°C. The experiment was conducted on fresh sweet sorghum. The results showed that the addition of the alpha amylase enzyme increased the content of reducing sugar and decreased levels of starch. Elevating concentration of alpha amylase enzyme will increase the reducing sugar content in sweet sorghum juice. The optimum alpha amylase enzyme concentration to produce the highest total sugar was 80 μL/100 mL of sweet sorghum juice with the optimum incubation time was 90 minutes. The results of this study are expected to create a new sweetener for sugar substitution. From the economic prospective aspect, sorghum is a potential crop and can be relied upon to support the success of the food diversification program which further leads to the world food security

  3. Cloning and expression of N-glycosylation-related glucosidase from Glaciozyma antarctica

    Science.gov (United States)

    Yajit, Noor Liana Mat; Kamaruddin, Shazilah; Hashim, Noor Haza Fazlin; Bakar, Farah Diba Abu; Murad, Abd. Munir Abd.; Mahadi, Nor Muhammad; Mackeen, Mukram Mohamed

    2016-11-01

    The need for functional oligosaccharides in various field is ever growing. The enzymatic approach for synthesis of oligosaccharides is advantageous over traditional chemical synthesis because of the regio- and stereo- selectivity that can be achieved without the need for protection chemistry. In this study, the α-glucosidase I protein sequence from Saccharomyces cerevisiae (UniProt database) was compared using Basic Local Alignment Search Tool (BLAST) with Glaciozyma antarctica genome database. Results showed 33% identity and an E-value of 1 × 10-125 for α-glucosidase I. The gene was amplified, cloned into the pPICZα C vector and used to transform Pichia pastoris X-33 cells. Soluble expression of α-Glucosidase I (˜91 kDa) was achieved at 28 °C with 1.0 % of methanol.

  4. Cloning, expression, and enzymatic activity evaluation of cholesterol oxidase gene isolated from a native Rhodococcus sp.

    Directory of Open Access Journals (Sweden)

    Hamed Esmaeil Lashgarian

    2016-10-01

    Full Text Available Cholesterol oxidase (CHO is one of the valuable enzymes that play an important role in: measurement of serum cholesterol, food industry as a biocatalyst and agriculture as a biological larvicide. This enzyme was produced by several bacterial strains. Wild type enzyme produced by Rhodococcus sp. secret two forms of CHO enzyme: extra cellular and membrane bound type which its amount is low and unstable. The goal of the study was cloning, expression, and enzymatic activity evaluation of cholesterol oxidase gene isolated from a native Rhodococcus sp. CHO gene was isolated from native bacteria and cloned into pET23a. In the next step, the construct was expressed in E.coli BL21 and induced by different concentration of IPTG ranges from 0.1 - 0.9 mM. This gene contains 1642 bp and encodes a protein consists of 533 amino acids. It has about 96 % homology with CHO gene isolated from Rhodococcus equi. The high expression was obtained in 0.5 mM concentration of IPTG after 4 hour induction. This recombinant enzyme had a molecular weight of 55 kDa, that secretion of intra cellular type is much more than extracellular form. The optimum pH and temperature conditions for the recombinant enzyme were 7.5 and 45°C, respectively. CHO enzyme obtained from Rhodococcus sp. is a cheap enzyme with medical and industrial applications that can be produced easily and purified in large scale with simple methods.

  5. Effect of Agromorphological Diversity and Botanical Race on Biochemical Composition in Sweet Grains Sorghum [Sorghum Bicolor (L. Moench] of Burkina Faso

    Directory of Open Access Journals (Sweden)

    Nerbéwendé Sawadogo

    2017-05-01

    Full Text Available Sorghum bicolor (L. Moench is an under-harvested crop in Burkina Faso. It is grown mainly for its sweet grains in the pasty stage. However, the precocity of the cycle and the sweet grains at pasty stage make it an interesting plant with agro-alimentary potential during the lean season. This study was carried out to identify the main sugars responsible for the sweetness of the grains at the pasty stage and their variation according to the agro-morphological group and the botanical race. Thus, the grains harvested at the pasty stage of fifteen (15 accessions selected according to the agro-morphological group and botanical race were lyophilized and analyzed by High Performance Liquid Chromatography (HPLC. The results reveal the presence of four (4 main carbohydrates at pasty stage of grains such as fructose, glucose, sucrose and starch. Analysis of variance revealed that these carbohydrates discriminate significantly the agro-morphological groups and the botanical races. Moreover, with exception of the sucrose, the coefficient of determination (R2 values shows that the agro-morphological group factor has a greater effect on the expression of glucose, fructose and starch than the botanical race. Group III and caudatum race have the highest levels of fructose and would be the sweetest. While group IV and the guinea-bicolor race with the low value of fructose would be the least sweet. Fructose is therefore the main sugar responsible for the sweetness of the pasty grains of sweet grains sorghum.

  6. Grain sorghum dust increases macromolecular efflux from the in situ nasal mucosa.

    Science.gov (United States)

    Gao, X P

    1998-04-01

    The purpose of this study was to determine whether an aqueous extract of grain sorghum dust increases macromolecular efflux from the nasal mucosa in vivo and, if so, whether this response is mediated, in part, by substance P. Suffusion of grain sorghum dust extract on the in situ nasal mucosa of anesthetized hamsters elicits a significant increase in clearance of fluorescein isothiocyanate-labeled dextran (FITC-dextran; mol mass, 70 kDa; P grain sorghum dust elicits neurogenic plasma exudation from the in situ nasal mucosa.

  7. Lipids characterization of ultrasound and microwave processed germinated sorghum.

    Science.gov (United States)

    Hassan, Sadia; Imran, Muhammad; Ahmad, Nazir; Khan, Muhammad Kamran

    2017-06-27

    Cereal crops and oilseeds provide diverse pool of fatty acids with characteristic properties. Sorghum (Sorghum bicolor (L.) Moench) provides the staple food with serving as main source of energy and protein. Germination of sorghum generally increases the nutritive value of seeds and the effects of germination on lipids composition of seeds vary greatly with processing conditions. Therefore, the current study was conducted to compare the effect of emerging processing techniques such as ultrasound (US) and microwave (MW) on fatty acids composition and oil yield of sorghum seeds before and after germination. Initially sorghum grains were soaked with 5% NaOCl (sodium hypochlorite) for surface sterilization. Afterwards, grains were soaked in excess water for 22 h at room temperature and were divided into four portions. The first portion (100 g grains) was subjected to germination without applying any microwave and ultrasonic treatment (T 0 ). Second portion was further divided into four groups (T 1 , T 2 , T 3 , T 4 ) (100 g of each group) and grains were subjected to ultrasonic treatments using two different ultrasonic intensities (US 1 : 40%; US 2 : 60%) within range of 0-100% and with two different time durations (t US1 : 5 min; t US2 : 10 min) at constant temperature. Third portion was also divided into four groups (T 1 , T 2 , T 3 , T 4 ) (100 g of each group) and exposed to microwave treatments at two different power levels (MW 1 : 450 watt; MW 2 : 700 watt) within the range of 100-900 W for two different time durations (t MW1 : 15 s; t MW2 : 30s). Similarly, fourth portion was divided into four groups (T 1 , T 2 , T 3 , T 4 ) (100 g of each group). Each group was exposed to both MW (MW 1 , MW 2 ) (100-900 watt power) & US (US 1 , US 2 ) (0-100% intensity) treatments at two different time levels (t US , t MW ). Then, germination was carried out and pre-treated raw and pre-treated germinated sorghum grains were analyzed for total oil yield, fatty acid

  8. Cloning and expression analysis of carboxyltransferase of acetyl-coA carboxylase from Jatropha curcas.

    Science.gov (United States)

    Xie, Wu-Wei; Gao, Shun; Wang, Sheng-Hua; Zhu, Jin-Qiu; Xu, Ying; Tang, Lin; Chen, Fang

    2010-01-01

    A full-length cDNA of the carboxyltransferase (accA) gene of acetyl-coenzym A (acetyl-CoA) carboxylase from Jatropha curcas was cloned and sequenced. The gene with an open reading frame (ORF) of 1149 bp encodes a polypeptide of 383 amino acids, with a molecular mass of 41.9 kDa. Utilizing fluorogenic real-time polymerase chain reaction (RT-PCR), the expression levels of the accA gene in leaves and fruits at early, middle and late stages under pH 7.0/8.0 and light/darkness stress were investigated. The expression levels of the accA gene in leaves at early, middle and late stages increased significantly under pH 8.0 stress compared to pH 7.0. Similarly, the expression levels in fruits showed a significant increase under darkness condition compared to the control. Under light stress, the expression levels in the fruits at early, middle and late stages showed the largest fluctuations compared to those of the control. These findings suggested that the expression levels of the accA gene are closely related to the growth conditions and developmental stages in the leaves and fruits of Jatropha curcas.

  9. Biolistic mediated sorghum (Sorghum bicolor L. Moench) transformation via mannose and bialaphos based selection systems

    CSIR Research Space (South Africa)

    Grootboom, AW

    2010-01-01

    Full Text Available of transformation. In sorghum, concerns about flow of herbicide and antibiotic resistance gene into genetically related wild and weedy species have a direct bearing on the choice of suitable selectable markers in many tropical and subtropical regions. The authors...

  10. Sorghum (Sorghum bicolor) varieties adopt strongly contrasting strategies in response to drought.

    Science.gov (United States)

    Ogbaga, Chukwuma C; Stepien, Piotr; Johnson, Giles N

    2014-10-01

    Sorghum is one of the most drought tolerant crops but surprisingly, little is known about the mechanisms achieving this. We have compared physiological and biochemical responses to drought in two sorghum cultivars with contrasting drought tolerance. These closely related cultivars have starkly contrasting responses to water deficit. In the less tolerant Samsorg 40, drought induced progressive loss of photosynthesis. The more drought tolerant Samsorg 17 maintained photosynthesis, transpiration and chlorophyll content until the most extreme conditions. In Samsorg 40, there was a highly specific down-regulation of selected proteins, with loss of PSII and Rubisco but maintenance of PSI and cytochrome b6 f, allowing plants to maintain ATP synthesis. The nitrogen released allows for accumulation of glycine betaine and proline. To the best of our knowledge, this is the first example of specific reengineering of the photosynthetic apparatus in response to drought. In contrast, in Samsorg 17 we detected no substantial change in the photosynthetic apparatus. Rather, plants showed constitutively high soluble sugar concentration, enabling them to maintain transpiration and photosynthesis, even in extremely dry conditions. The implications for these strikingly contrasted strategies are discussed in relation to agricultural and natural systems. © 2014 Scandinavian Plant Physiology Society.

  11. Cloning and Expression of TRYP6 Gene from Leishmania major (MRHO/IR/75/ER

    Directory of Open Access Journals (Sweden)

    G Eslami

    2008-06-01

    Full Text Available Background: Leishmania, needs to detoxify the macrophage derived potent peroxides (H2O2. Tryparedoxin path­way contains tryparedoxin peroxidase (TSA or TRYP. The aim of the study was to detect the full-length gene se­quence and its encoded protein of the LmTRYP6 gene (EU251502, and comparison the gene sequence with LmTRYP6 (LmjF15.1140, another previously reported member of this gene family.Methods: L.major (MRHO/IR/75/ER promastigotes were cultured, DNA and RNA were extracted and the inter­ested gene was amplified using PCR and RT-PCR methods.  PCR/ RT-PCR fragments were purified and cloned first in pTZ57R/T and then in pET15b expression vector. The expressed protein was verified using western blot method. Char­acterization of the expressed protein was performed bioinformatically.Results: Molecular evaluation revealed that the cloned LmTRYP6 gene (EU251502 encoded a predicted 184 amino acid long protein with a theoretical isoelectric point of 6.1101. Alignment showed a number of changes in amino acid composition including the replacement of highly conserved Trp177 by Cys in LmTRYP6 (ABX26130.Conclusion: So far no study has been done on this group, i.e.  TRYP6 gene, from tryparedoxin peroxidase family. The low homology with LmTRYP6 (LmjF15.1140 and vast array of differences observed in the gene under study (LmTRYP6; EU251502 could open new windows in the field of anti-Leishmania combat. Based on its important role in the viability and successful establishment of the parasite in the host organism it looks to be very good candi­date for vaccine development and any other sort of novel drug development.

  12. Phenolic compounds and related enzymes as determinants of sorghum for food use

    NARCIS (Netherlands)

    Dicko, M.H.; Gruppen, H.; Traore, A.S.; Voragen, A.G.J.; Berkel, van W.J.H.

    2006-01-01

    Phenolic compounds and related enzymes such as phenol biosynthesizing enzymes (phenylalanine ammonia lyase) and phenol catabolizing enzymes (polyphenol oxidase and peroxidase) are determinants for sorghum utilization as human food because they influence product properties during and after sorghum

  13. Cloning, sequence analysis, and expression of the large subunit of the human lymphocyte activation antigen 4F2

    International Nuclear Information System (INIS)

    Lumadue, J.A.; Glick, A.B.; Ruddle, F.H.

    1987-01-01

    Among the earliest expressed antigens on the surface of activated human lymphocytes is the surface antigen 4F2. The authors have used DNA-mediated gene transfer and fluorescence-activated cell sorting to obtain cell lines that contain the gene encoding the large subunit of the human 4F2 antigen in a mouse L-cell background. Human DNAs cloned from these cell lines were subsequently used as hybridization probes to isolate a full-length cDNA clone expressing 4F2. Sequence analysis of the coding region has revealed an amino acid sequence of 529 residues. Hydrophobicity plotting has predicted a probable structure for the protein that includes an external carboxyl terminus, an internal leader sequence, a single hydrophobic transmembrane domain, and two possible membrane-associated domains. The 4F2 cDNA detects a single 1.8-kilobase mRNA in T-cell and B-cell lines. RNA gel blot analysis of RNA derived from quiescent and serum-stimulated Swiss 3T3 fibroblasts reveals a cell-cycle modulation of 4F2 gene expression: the mRNA is present in quiescent fibroblasts but increases 8-fold 24-36 hr after stimulation, at the time of maximal DNA synthesis

  14. Cloning, sequence analysis, and expression of the large subunit of the human lymphocyte activation antigen 4F2

    Energy Technology Data Exchange (ETDEWEB)

    Lumadue, J.A.; Glick, A.B.; Ruddle, F.H.

    1987-12-01

    Among the earliest expressed antigens on the surface of activated human lymphocytes is the surface antigen 4F2. The authors have used DNA-mediated gene transfer and fluorescence-activated cell sorting to obtain cell lines that contain the gene encoding the large subunit of the human 4F2 antigen in a mouse L-cell background. Human DNAs cloned from these cell lines were subsequently used as hybridization probes to isolate a full-length cDNA clone expressing 4F2. Sequence analysis of the coding region has revealed an amino acid sequence of 529 residues. Hydrophobicity plotting has predicted a probable structure for the protein that includes an external carboxyl terminus, an internal leader sequence, a single hydrophobic transmembrane domain, and two possible membrane-associated domains. The 4F2 cDNA detects a single 1.8-kilobase mRNA in T-cell and B-cell lines. RNA gel blot analysis of RNA derived from quiescent and serum-stimulated Swiss 3T3 fibroblasts reveals a cell-cycle modulation of 4F2 gene expression: the mRNA is present in quiescent fibroblasts but increases 8-fold 24-36 hr after stimulation, at the time of maximal DNA synthesis.

  15. Effect of sorghum type and malting on production of free amino nitrogen in conjunction with exogenous protease enzymes.

    Science.gov (United States)

    Dlamini, Bhekisisa C; Buys, Elna M; Taylor, John R N

    2015-01-01

    Sorghum types suitable for brewing and bioethanol production are required. The effect of sorghum type (white non-tannin versus white type II tannin) on free amino nitrogen (FAN) production from sorghum grain and malt using exogenous protease enzymes was investigated over extended incubation at moderate temperature (45 °C). With grain in the absence of exogenous proteases, white non-tannin sorghum produced substantially higher levels of FAN than white type II tannin sorghum, due to the tannins in the latter. Incubating sorghum grain with neutral proteinase and amino-peptidase in combination improved FAN production. The two sorghum types produced similar FAN levels when malted and incubated in the absence of the exogenous proteases. When both sorghums were malted and incubated with neutral proteinase alone substantially more FAN yield (124-126 mg 100 g(-1)) occurred than with grains (61-84 mg 100 g(-1)). The combination of amino-peptidase and proteinase did not improve FAN further. Neither, did malting influence wort free amino acid profile. Group B amino acids constituted the highest percentage (42-47%). With grain, white non-tannin sorghum plus proteinase and amino-peptidase yields the highest FAN, with malt both white non-tannin and white type II tannin sorghums plus proteinase yield the highest FAN. © 2014 Society of Chemical Industry.

  16. Critical Factors Affecting the Success of Cloning, Expression, and Mass Production of Enzymes by Recombinant E. coli.

    Science.gov (United States)

    Fakruddin, Md; Mohammad Mazumdar, Reaz; Bin Mannan, Khanjada Shahnewaj; Chowdhury, Abhijit; Hossain, Md Nur

    2013-01-01

    E. coli is the most frequently used host for production of enzymes and other proteins by recombinant DNA technology. E. coli is preferable for its relative simplicity, inexpensive and fast high-density cultivation, well-known genetics, and large number of compatible molecular tools available. Despite all these advantages, expression and production of recombinant enzymes are not always successful and often result in insoluble and nonfunctional proteins. There are many factors that affect the success of cloning, expression, and mass production of enzymes by recombinant E. coli. In this paper, these critical factors and approaches to overcome these obstacles are summarized focusing controlled expression of target protein/enzyme in an unmodified form at industrial level.

  17. Arsenic-contaminated soils. Phytotoxicity studies with sunflower and sorghum

    Energy Technology Data Exchange (ETDEWEB)

    Lyubun, Y.V.; Kosterin, P.V.; Zakharova, E.A.; Fedorov, E.E. [Inst. of Biochemistry and Physiology of Plants and Microorganisms, Russian Academy of Sciences, Saratov (Russian Federation); Shcherbakov, A.A. [Saratov Military Inst. of Radiological, Chemical and Biological Defence, Saratov (Russian Federation)

    2002-07-01

    Background, Aim and Scope. Environmental pollution caused by arsenic (As) is a major ecological problem. There has been intense worldwide effort to find As-hyperaccumulating plants that can be used in phytoremediation - the green-plant-assisted removal of chemical pollutants from soils. For phytoremediation, it is natural to prefer cultivated rather than wild plants, because their agriculture is well known. This study was conducted to evaluate the tolerance of common sunflower (Helianthus annuus L.) and sugar sorghum (Sorghum saccharatum Pers.) for soil-As contents of 10-100 mg As kg{sup -1} soil, with sodium arsenite as a model contaminant. Methods. Plants were grown in a growth chamber for 30 days. Microfield experiments were conducted on experimental plots. To study the phytoremediation effect of the auxins indole-3-acetic acid (IAA) and 2,4-dichlorophenoxyacetic acid (2,4-D), we treated 1- and 3-day-old plant seedlings with water solutions of the auxins (concentrations of 10{sup -5}, 10{sup -7}, and 10{sup -9} g l{sup -1}). The soil and plant-biomass samples were analyzed for total As by using the color reaction of ammonium molybdate with As. Results and Discussion. Phytotoxicity studies showed that 100 mg as kg{sup -1} soil poisoned sunflower and sorghum growth by 50%. There was a linear correlation between soil-As content and As accumulation in the plants. Laboratory experiments showed that the soil-As content was reduced two- to threefold after sunflower had been grown with 10-100 mg As kg{sup -1} soil for 30 days. Treatment of sunflower and sorghum seedlings with IAA and 2,4-D at a concentration of 10{sup -5} g l{sup -1} in microfield experiments enhanced the phytoremediation two- to fivefold as compared with untreated control plants. The best results were obtained with 3-day-old seedlings. Conclusion, Recommendation and Outlook. (a) Sunflower and sorghum are good candidates to remediate As-polluted soils. (b) Phytoremediation can be improved with IAA or 2

  18. Diversity, users' perception and food processing of sorghum: implications for dietary iron and zinc supply

    NARCIS (Netherlands)

    Kayodé, A.P.P.

    2006-01-01

    This thesis focuses on the diversity of sorghum and its post-harvest processing into food. We studied the contribution that sorghum can make to Fe and Zn intake by poor people in Africa, using the situation in Benin as a study context. The culinary and sensory characteristics of sorghum crops and

  19. Assessment of N2 fixing efficiency of Beijerinckia indica and Azospirillum brasilense in Sorghum (Sorghum bicolor (L.) moench) using 15N tracer

    International Nuclear Information System (INIS)

    Kanimoli, S.; Marimuthu, P.; Arulmozhiselvan, K.

    2010-01-01

    For studying the benefits of inoculation of N 2 fixing diazotrophs in the root zone of sorghum crop, a pot culture was conducted on neutral red sandy loam soil with sorghum cv. CO26, using 15 N tracer. At the end of 45 days duration after sowing, Beijerinckia indica inoculation contributed 56.9 per cent N derived from N 2 fixation, out of total N concentration in whole drymatter of sorghum plant. It proved to be the efficient N 2 fixer by contributing N from N 2 fixation to the tune of 17.6 Kg -1 . Accumulation of N derived from N 2 fixation from B. indica was primarily in leaf blade (50.0%) followed by stem (31.8%), leaf sheath (14.0%) and root (4.2%). Inoculation of Azospirillum brasllense accelerated uptake of N from soil and fertilizer N sources compared to B. indica and hence registered low N fixation. (author)

  20. Purification and characterization of recombinant human bile salt-stimulated lipase expressed in milk of transgenic cloned cows

    Science.gov (United States)

    Ding, Fangrong; Wang, Tao; Liu, Wenjie; Lindquist, Susanne; Hernell, Olle; Wang, Jianwu; Li, Jing; Li, Ling; Zhao, Yaofeng; Dai, Yunping; Li, Ning

    2017-01-01

    Bile salt-stimulated lipase (BSSL) is a lipolytic digestive enzyme with broad substrate specificity secreted from exocrine pancreas into the intestinal lumen in all species and from the lactating mammary gland into the milk of some species, notably humans but not cows. BSSL in breast milk facilitates digestion and absorption of milk fat and promotes growth of small for gestational age preterm infants. Thus, purified recombinant human BSSL (rhBSSL) can be used for treatment of patients with fat malabsorption and expressing rhBSSL in the milk of transgenic cloned cows would therefore be a mean to meet a medical need. In the present study, a vector pBAC-hLF-hBSSL was constructed, which efficiently expressed active rhBSSL in milk of transgenic cloned cows to a concentration of 9.8 mg/ml. The rhBSSL purified from cow milk had the same enzymatic activity, N-terminal amino acid sequence, amino acid composition and isoelectric point and similar physicochemical characteristics as human native BSSL. Our study supports the use of transgenic cattle for the cost-competitive, large-scale production of therapeutic rhBSSL. PMID:28475629

  1. Using elevated CO2 to increase the biomass of a Sorghum vulgare x Sorghum vulgare var. sudanense hybrid and Trifolium pratense L. and to trigger hyperaccumulation of cesium

    International Nuclear Information System (INIS)

    Wu Huibin; Tang Shirong; Zhang Ximei; Guo Junkang; Song, Zhengguo; Tian Shuai; Smith, Donald L.

    2009-01-01

    The most important challenge to use phytoremediation is how to improve its efficiency by increasing the accumulation of metals in plants, or by improving key plant biological traits that should enhance metal uptake. In this paper, we used open-top chambers to investigate the effects of elevated CO 2 (860 μL L -1 ) on biomass and Cs uptake by a Sorghum vulgare x Sorghum vulgare var. sudanense hybrid and Trifolium pratense L. growing on soils spiked with various levels of cesium (0, 300, 1500 and 3000 mg Cs kg -1 ). The results showed that elevated CO 2 not only increased aboveground biomass of the Sorghum and Trifolium species by 32-111%, and by 8-11%, respectively, compared to the ambient CO 2 treatment, but also caused more accumulation of Cs by Sorghum species (up to 73%) than Trifolium species (up to 43%). It was speculated that the increase in biomass and the improvement in Cs accumulation ability at elevated CO 2 could be related to lowered soil pH values, and changes in number and kind of microorganisms in the rhizospheres of the two tested species. This is the first report of a link among elevated CO 2 , increased biomass and hyperaccumulation of Cs by Sorghum and Trifolium species.

  2. Quantum dot-based molecular imaging of cancer cell growth using a clone formation assay.

    Science.gov (United States)

    Geng, Xia-Fei; Fang, Min; Liu, Shao-Ping; Li, Yan

    2016-10-01

    This aim of the present study was to investigate clonal growth behavior and analyze the proliferation characteristics of cancer cells. The MCF‑7 human breast cancer cell line, SW480 human colon cancer cell line and SGC7901 human gastric cancer cell line were selected to investigate the morphology of cell clones. Quantum dot‑based molecular targeted imaging techniques (which stained pan‑cytokeratin in the cytoplasm green and Ki67 in the cell nucleus yellow or red) were used to investigate the clone formation rate, cell morphology, discrete tendency, and Ki67 expression and distribution in clones. From the cell clone formation assay, the MCF‑7, SW480 and SGC7901 cells were observed to form clones on days 6, 8 and 12 of cell culture, respectively. These three types of cells had heterogeneous morphology, large nuclear:cytoplasmic ratios, and conspicuous pathological mitotic features. The cells at the clone periphery formed multiple pseudopodium. In certain clones, cancer cells at the borderline were separated from the central cell clusters or presented a discrete tendency. With quantum dot‑based molecular targeted imaging techniques, cells with strong Ki67 expression were predominantly shown to be distributed at the clone periphery, or concentrated on one side of the clones. In conclusion, cancer cell clones showed asymmetric growth behavior, and Ki67 was widely expressed in clones of these three cell lines, with strong expression around the clones, or aggregated at one side. Cell clone formation assay based on quantum dots molecular imaging offered a novel method to study the proliferative features of cancer cells, thus providing a further insight into tumor biology.

  3. Sorghum and rice: Mali

    International Nuclear Information System (INIS)

    2003-01-01

    Agriculture is the mainstay of the Malian economy and yet cereal imports absorb 6.5% of GDP. Food self-sufficiency is therefore a national priority. The Joint FAO/IAEA Division is supporting a programme to improve local varieties of sorghum and rice by using nuclear techniques to develop new cultivars that will produce higher yields under Mali's semi-arid climatic conditions. (IAEA)

  4. The influence of time and severity of Striga infection on the Sorghum bicolor - Striga hermonthica association

    NARCIS (Netherlands)

    Ast, van A.

    2006-01-01

    Keywords: Striga hermonthica , Sorghum bicolor , infection time, infection level, tolerance.This thesis presents the results of a study on the interaction between the parasitic weed Strigahermonthica (Del.) Benth. and sorghum ( Sorghum bicolor [L.] Moench). The main objective of the study was

  5. Molecular cloning, expression, and characterization of mouse amine N-sulfotransferases

    International Nuclear Information System (INIS)

    Takahashi, Saki; Sakakibara, Yoichi; Mishiro, Emi; Kouriki, Haruna; Nobe, Rika; Kurogi, Katsuhisa; Yasuda, Shin; Liu, M.-C.; Suiko, Masahito

    2008-01-01

    By searching the GenBank database, we recently identified a novel mouse cytosolic sulfotransferase (SULT) cDNA (IMAGE Clone ID 679629) and a novel mouse SULT gene (LOC 215895). Sequence analysis revealed that both mouse SULTs belong to the cytosolic SULT3 gene family. The recombinant form of these two newly identified SULTs, designated SULT3A1 and SULT3A2, were expressed using the pGEX-4T-1 glutathione S-transferase fusion system and purified from transformed BL21 Escherichia coli cells. Both purified SULT3A1 and SULT3A2 exhibited strong amine N-sulfonating activities toward 1-naphthylamine among a variety of endogenous and xenobiotic compounds tested as substrates. Kinetic constants of the sulfation of 1-naphthylamine and 1-naphthol by these two enzymes were determined. Collectively, these results imply that these two amine-sulfonating SULT3s may play essential roles in the metabolism and detoxification of aromatic amine compounds in the body

  6. Sorghum grain as human food in Africa: relevance of content of ...

    African Journals Online (AJOL)

    Sorghum is a staple food grain in many semi-arid and tropic areas of the world, notably in Sub-Saharan Africa because of its good adaptation to hard environments and its good yield of production. Among important biochemical components for sorghum processing are levels of starch (amylose and amylopectin) and starch ...

  7. A Gateway MultiSite recombination cloning toolkit.

    Directory of Open Access Journals (Sweden)

    Lena K Petersen

    Full Text Available The generation of DNA constructs is often a rate-limiting step in conducting biological experiments. Recombination cloning of single DNA fragments using the Gateway system provided an advance over traditional restriction enzyme cloning due to increases in efficiency and reliability. Here we introduce a series of entry clones and a destination vector for use in two, three, and four fragment Gateway MultiSite recombination cloning whose advantages include increased flexibility and versatility. In contrast to Gateway single-fragment cloning approaches where variations are typically incorporated into model system-specific destination vectors, our Gateway MultiSite cloning strategy incorporates variations in easily generated entry clones that are model system-independent. In particular, we present entry clones containing insertions of GAL4, QF, UAS, QUAS, eGFP, and mCherry, among others, and demonstrate their in vivo functionality in Drosophila by using them to generate expression clones including GAL4 and QF drivers for various trp ion channel family members, UAS and QUAS excitatory and inhibitory light-gated ion channels, and QUAS red and green fluorescent synaptic vesicle markers. We thus establish a starter toolkit of modular Gateway MultiSite entry clones potentially adaptable to any model system. An inventory of entry clones and destination vectors for Gateway MultiSite cloning has also been established (www.gatewaymultisite.org.

  8. Heterogeneity of functional properties of Clone 66 murine breast cancer cells expressing various stem cell phenotypes.

    Science.gov (United States)

    Mukhopadhyay, Partha; Farrell, Tracy; Sharma, Gayatri; McGuire, Timothy R; O'Kane, Barbara; Sharp, J Graham

    2013-01-01

    Breast cancer grows, metastasizes and relapses from rare, therapy resistant cells with a stem cell phenotype (cancer stem cells/CSCs). However, there is a lack of studies comparing the functions of CSCs isolated using different phenotypes in order to determine if CSCs are homogeneous or heterogeneous. Cells with various stem cell phenotypes were isolated by sorting from Clone 66 murine breast cancer cells that grow orthotopically in immune intact syngeneic mice. These populations were compared by in vitro functional assays for proliferation, growth, sphere and colony formation; and in vivo limiting dilution analysis of tumorigenesis. The proportion of cells expressing CD44(high)CD24(low/neg), side population (SP) cells, ALDH1(+), CD49f(high), CD133(high), and CD34(high) differed, suggesting heterogeneity. Differences in frequency and size of tumor spheres from these populations were observed. Higher rates of proliferation of non-SP, ALDH1(+), CD34(low), and CD49f(high) suggested properties of transit amplifying cells. Colony formation was higher from ALDH1(-) and non-SP cells than ALDH1(+) and SP cells suggesting a progenitor phenotype. The frequency of clonal colonies that grew in agar varied and was differentially altered by the presence of Matrigel™. In vivo, fewer cells with a stem cell phenotype were needed for tumor formation than "non-stem" cells. Fewer SP cells were needed to form tumors than ALDH1(+) cells suggesting further heterogeneities of cells with stem phenotypes. Different levels of cytokines/chemokines were produced by Clone 66 with RANTES being the highest. Whether the heterogeneity reflects soluble factor production remains to be determined. These data demonstrate that Clone 66 murine breast cancer cells that express stem cell phenotypes are heterogeneous and exhibit different functional properties, and this may also be the case for human breast cancer stem cells.

  9. Establishment and characterization of a spontaneously immortalized trophoblast cell line (HPT-8) and its hepatitis B virus-expressing clone.

    Science.gov (United States)

    Zhang, Lei; Zhang, Weilu; Shao, Chen; Zhang, Jingxia; Men, Ke; Shao, Zhongjun; Yan, Yongping; Xu, Dezhong

    2011-08-01

    Most trophoblast cell lines currently available to study vertical transmission of hepatitis B virus (HBV) are immortalized by viral transformation. Our goal was to establish and characterize a spontaneously immortalized human first-trimester trophoblast cell line and its HBV-expressing clone. Chorionic villi of Asian human first-trimester placentae were digested with trypsin and collagenase I to obtain the primary trophoblast cell culture. A spontaneously immortalized trophoblast cell line (HPT-8) was analyzed by scanning and transmission electron microscopy, cell cycle analysis, immunohistochemistry and immunofluorescence. HPT-8 cells were stably transfected with the adr subtype of HBV (HPT-8-HBV) and characterized by PCR and enzyme-linked immunosorbent assay. We obtained a clonal derivative of a spontaneously immortalized primary cell clone (HPT-8). HPT-8 cells were epithelioid and polygonal, and formed multinucleate, giant cells. They exhibited microvilli, distinct desmosomes between adjacent cells, abundant endoplasm, lipid inclusions and glycogen granules, which are all characteristic of cytotrophoblasts. HPT-8 cells expressed cytokeratin 7, cytokeratin 18, vimentin, cluster of differentiation antigen 9, epidermal growth factor receptor, stromal cell-derived factor 1 and placental alkaline phosphatase. They secreted prolactin, estradiol, progesterone and hCG, and were positive for HLA-G, a marker of extravillous trophoblasts. HPT-8-HBV cells were positive for HBV relaxed-circular, covalently closed circular DNA and pre-S sequence. HPT-8-HBV cells also produced and secreted HBV surface antigen and HBV e antigen. We established a trophoblast cell line, HPT-8 and its HBV-expressing clone which could be valuable in exploring the mechanism of HBV viral integration in human trophoblasts during intrauterine infection.

  10. Development of a gene cloning system in a fast-growing and moderately thermophilic Streptomyces species and heterologous expression of Streptomyces antibiotic biosynthetic gene clusters

    Science.gov (United States)

    2011-01-01

    Background Streptomyces species are a major source of antibiotics. They usually grow slowly at their optimal temperature and fermentation of industrial strains in a large scale often takes a long time, consuming more energy and materials than some other bacterial industrial strains (e.g., E. coli and Bacillus). Most thermophilic Streptomyces species grow fast, but no gene cloning systems have been developed in such strains. Results We report here the isolation of 41 fast-growing (about twice the rate of S. coelicolor), moderately thermophilic (growing at both 30°C and 50°C) Streptomyces strains, detection of one linear and three circular plasmids in them, and sequencing of a 6996-bp plasmid, pTSC1, from one of them. pTSC1-derived pCWH1 could replicate in both thermophilic and mesophilic Streptomyces strains. On the other hand, several Streptomyces replicons function in thermophilic Streptomyces species. By examining ten well-sporulating strains, we found two promising cloning hosts, 2C and 4F. A gene cloning system was established by using the two strains. The actinorhodin and anthramycin biosynthetic gene clusters from mesophilic S. coelicolor A3(2) and thermophilic S. refuineus were heterologously expressed in one of the hosts. Conclusions We have developed a gene cloning and expression system in a fast-growing and moderately thermophilic Streptomyces species. Although just a few plasmids and one antibiotic biosynthetic gene cluster from mesophilic Streptomyces were successfully expressed in thermophilic Streptomyces species, we expect that by utilizing thermophilic Streptomyces-specific promoters, more genes and especially antibiotic genes clusters of mesophilic Streptomyces should be heterologously expressed. PMID:22032628

  11. Generation of a vector system facilitating cloning of DMBT1 variants and recombinant expression of functional full-length DMBT1

    DEFF Research Database (Denmark)

    End, Caroline; Lyer, Stefan; Renner, Marcus

    2005-01-01

    of a vector system that facilitates cloning of DMBT1 variants. We demonstrate applicability of the vector system by expression of the largest DMBT1 variant in a tetracycline-inducible mammalian expression system using the Chinese hamster ovary cell line. Yields up to 30 mg rDMBT1 per litre of cell culture......Deleted in malignant brain tumours 1 (DMBT1) codes for a approximately 340kDa glycoprotein with highly repetitive scavenger receptor cysteine-rich (SRCR) domains. DMBT1 was implicated in cancer, defence against viral and bacterial infections, and differentiation of epithelial cells. Recombinant...... yields, and protein preparations which may substantially vary due to differential processing and genetic polymorphism, all of which impedes functional research on DMBT1. Cloning of DMBT1 cDNAs is hampered because of the size and the 13 highly homologous SRCR exons. In this study, we report on the setup...

  12. Molecular cloning and expression analysis of jasmonic acid dependent but salicylic acid independent LeWRKY1.

    Science.gov (United States)

    Lu, M; Wang, L F; Du, X H; Yu, Y K; Pan, J B; Nan, Z J; Han, J; Wang, W X; Zhang, Q Z; Sun, Q P

    2015-11-30

    Various plant genes can be activated or inhibited by phytohormones under conditions of biotic and abiotic stress, especially in response to jasmonic acid (JA) and salicylic acid (SA). Interactions between JA and SA may be synergistic or antagonistic, depending on the stress condition. In this study, we cloned a full-length cDNA (LeWRKY1, GenBank accession No. FJ654265) from Lycopersicon esculentum by rapid amplification of cDNA ends. Sequence analysis showed that this gene is a group II WRKY transcription factor. Analysis of LeWRKY1 mRNA expression in various tissues by qRT-PCR showed that the highest and lowest expression occurred in the leaves and stems, respectively. In addition, LeWRKY1 expression was induced by JA and Botrytis cinerea Pers., but not by SA.

  13. Nutrient content of sorghum beer strainings

    African Journals Online (AJOL)

    Sorghum beer strainings were analysed for starch, protein, fat, crude fibre, ash, minerals and ... The importance of minerals in animal nutrition has been recognized for many ..... strainings is probably due to yeast activity during fermentation ...

  14. Cloning, expression and characterization of phenylalanine ammonia-lyase from Rhodotorula glutinis.

    Science.gov (United States)

    Zhu, Longbao; Cui, Wenjing; Fang, Yueqin; Liu, Yi; Gao, Xinxing; Zhou, Zhemin

    2013-05-01

    The industrial-scale production of phenylalanine ammonia-lyase (PAL) mainly uses strains of Rhodotorula. However, the PAL gene from Rhodotorula has not been cloned. Here, the full-length gene of PAL from Rhodotorula glutinis was isolated. It was 2,121 bp, encoding a polypeptide with 706 amino acids and a calculated MW of 75.5 kDa. Though R. glutinis is an anamorph of Rhodosporium toruloides, the amino acid sequences of PALs them are not the same (about 74 % identity). PAL was expressed in E. coli and characterized. Its specific activity was 4.2 U mg(-1) and the k cat/K m was 1.9 × 10(4) mM(-1) s(-1), exhibiting the highest catalytic ability among the reported PALs. The genetic and biochemical information reported here should facilitate future application in industry.

  15. Lodging markedly reduced the biomass of sweet sorghum via decreasing photosynthesis in saline-alkali field

    Science.gov (United States)

    Guo, Jian Rong; Fan, Hai; Wang, Bao Shan

    2018-06-01

    Lodging is a serious problem in plant growth, especially in crops growth of the natural habitat. In order to determine the influence of lodging on the growth characters of sweet sorghum, plants grown in natural saline-alkali environment were used to investigate the fresh weight, dry weight, sugar content in the stalks and the photosynthesis index of salt tolerant crop sweet sorghum. Results showed that lodging significantly reduced the growth of sweet sorghum, the fresh weight and dry weight was only 28.3% and 22.5% of the normal plants when lodging occurred after 49 days. Lodging also reduced the stalks sugar content of sweet sorghum, the stalk sugar content of lodged plants was only 45.4% of that in the normal plants, when lodging occurred for 49 days. Lodging reduced the growth and sugar content by reducing the photosynthesis parameters of sweet sorghum grown in the saline-alkali field, thus, affected the accumulation of photosynthate. Interestingly, with the extension of the lodging time, lodging led to a decrease in photosynthetic rate of sweet sorghum mainly due to non-stomatal factors.

  16. Molecular cloning and expression analysis of KIN10 and cold-acclimation related genes in wild banana 'Huanxi' (Musa itinerans).

    Science.gov (United States)

    Liu, Weihua; Cheng, Chunzhen; Lai, Gongti; Lin, Yuling; Lai, Zhongxiong

    2015-01-01

    Banana cultivars may experience chilling or freezing injury in some of their cultivated regions, where wild banana can still grow very well. The clarification of the cold-resistant mechanism of wild banana is vital for cold-resistant banana breeding. In this study, the central stress integrator gene KIN10 and some cold-acclimation related genes (HOS1 and ICE1s) from the cold-resistant wild banana 'Huanxi' (Musa itinerans) were cloned and their expression patterns under different temperature treatments were analyzed. Thirteen full-length cDNA transcripts including 6 KIN10s, 1 HOS1 and 6 ICE1s were successfully cloned. Quantitative real-time PCR (qRT-PCR) results showed that all these genes had the highest expression levels at the critical temperature of banana (13 °C). Under chilling temperature (4 °C), the expression level of KIN10 reduced significantly but the expression of HOS1 was still higher than that at the optimal temperature (28 °C, control). Both KIN10 and HOS1 showed the lowest expression levels at 0 °C, the expression level of ICE1, however, was higher than control. As sucrose plays role in plant cold-acclimation and in regulation of KIN10 and HOS1 bioactivities, the sucrose contents of wild banana under different temperatures were detected. Results showed that the sucrose content increased as temperature lowered. Our result suggested that KIN10 may participate in cold stress response via regulating sucrose biosynthesis, which is helpful in regulating cold acclimation pathway in wild banana.

  17. Soil Organic Carbon Response to Cover Crop and Nitrogen Fertilization under Bioenergy Sorghum

    Science.gov (United States)

    Sainju, U. M.; Singh, H. P.; Singh, B. P.

    2015-12-01

    Removal of aboveground biomass for bioenergy/feedstock in bioenergy cropping systems may reduce soil C storage. Cover crop and N fertilization may provide additional crop residue C and sustain soil C storage compared with no cover crop and N fertilization. We evaluated the effect of four winter cover crops (control or no cover crop, cereal rye, hairy vetch, and hairy vetch/cereal rye mixture) and two N fertilization rates (0 and 90 kg N ha-1) on soil organic C (SOC) at 0-5, 5-15, and 15-30 cm depths under forage and sweet sorghums from 2010 to 2013 in Fort Valley, GA. Cover crop biomass yield and C content were greater with vetch/rye mixture than vetch or rye alone and the control, regardless of sorghum species. Soil organic C was greater with vetch/rye than rye at 0-5 and 15-30 cm in 2011 and 2013 and greater with vetch than rye at 5-15 cm in 2011 under forage sorghum. Under sweet sorghum, SOC was greater with cover crops than the control at 0-5 cm, but greater with vetch and the control than vetch/rye at 15-30 cm. The SOC increased at the rates of 0.30 Mg C ha-1 yr-1 at 0-5 cm for rye and the control to 1.44 Mg C ha-1 yr-1 at 15-30 cm for vetch/rye and the control from 2010 to 2013 under forage sorghum. Under sweet sorghum, SOC also increased linearly at all depths from 2010 to 2013, regardless of cover crops. Nitrogen fertilization had little effect on SOC. Cover crops increased soil C storage compared with no cover crop due to greater crop residue C returned to the soil under forage and sweet sorghum and hairy vetch/cereal rye mixture had greater C storage than other cover crops under forage sorghum.

  18. Statistical screening and selection of sweet sorghum varieties for bioethanol production

    International Nuclear Information System (INIS)

    Mehmood, S.; Aqil, T.; Tahir, M.S.

    2014-01-01

    This study aims at the screening of four cultivars of sorghums as a feedstock for bioethanol production. The straw of these varieties were subjected to pretreatment (dilute sulfuric acid) followed by enzyme hydrolysis to evaluate their potential to produce sugars. Four factor full factorial experimental design (2*2*2*4=32) was used to investigate the effects of experimental factors; sorghum varieties (84-Y-01, 85-G-86, Mr. Buster and RARI S-3), acid concentration (1 and 2%), temperature (121 and 140 degree C) and pretreatment time (30 and 60 min). The tested sorghum varieties follow the order 85-G-86 (47 g/100g) > Mr. Buster (44.6 g/100g) > 84-Y-01 (42 g/100g) > RARI S-3 (36 g/100g) for their sugar yield. The factors followed given order of significance; variety > temperature > acid concentration > pretreatment time. Sorghum variety (85-G-86) was selected as an appropriate feedstock for bioethanol production due to its higher sugar yield and lower concentration of by-products and furans. (author)

  19. Growing sweet sorghum as a source of fermentable sugars for energy

    Energy Technology Data Exchange (ETDEWEB)

    Gascho, G.J.; Nichols, R.L.; Powell-Gaines, T.

    1984-08-01

    Studies were undertaken on the southern coastal plain (Georgia) of the USA on sweet sorghum to evaluate its potential as a fuel ethanol feedstock. Field experiments were designed over three years to study several aspects of the production of fermentable sugars from sweet sorghum and these included cultivar types, fertility needs, weed control and growth regulation. Wray was the best cultivar, producing a high sugar per hectare. To justify the operation of an ethanol plant, sweet sorghum should be harvested over a period of months, so cultivars were selected for yearly, medium and late maturity, thus ensuring a constant supply of feedstock over a four month period. The fertility needs of sweet sorghum appear to be relatively low and the yield response to applications of N, P, K are given. The best weed control was achieved by treating with Propazine plus Metolacheor. Application of several growth regulators such as Gibberellin didn't significantly increase the yield of sugars. Finally, a method to measure the fermentable sugars was developd using the Technicon Autoanalyser II.

  20. Isolation, cDNA cloning and gene expression of an antibacterial protein from larvae of the coconut rhinoceros beetle, Oryctes rhinoceros.

    Science.gov (United States)

    Yang, J; Yamamoto, M; Ishibashi, J; Taniai, K; Yamakawa, M

    1998-08-01

    An antibacterial protein, designated rhinocerosin, was purified to homogeneity from larvae of the coconut rhinoceros beetle, Oryctes rhinoceros immunized with Escherichia coli. Based on the amino acid sequence of the N-terminal region, a degenerate primer was synthesized and reverse-transcriptase PCR was performed to clone rhinocerosin cDNA. As a result, a 279-bp fragment was obtained. The complete nucleotide sequence was determined by sequencing the extended rhinocerosin cDNA clone by 5' rapid amplification of cDNA ends. The deduced amino acid sequence of the mature portion of rhinocerosin was composed of 72 amino acids without cystein residues and was shown to be rich in glycine (11.1%) and proline (11.1%) residues. Comparison of the deduced amino acid sequence of rhinocerosin with those of other antibacterial proteins indicated that it has 77.8% and 44.6% identity with holotricin 2 and coleoptrecin, respectively. Rhinocerosin had strong antibacterial activity against E. coli, Streptococcus pyogenes, Staphylococcus aureus but not against Pseudomonas aeruginosa. Results of reverse-transcriptase PCR analysis of gene expression in different tissues indicated that the rhinocerosin gene is strongly expressed in the fat body and the Malpighian tubule, and weakly expressed in hemocytes and midgut. In addition, gene expression was inducible by bacteria in the fat body, the Malpighian tubule and hemocyte but constitutive expression was observed in the midgut.

  1. Problems, control, and opportunity of starch in the large scale processing of sugarcane and sweet sorghum

    Science.gov (United States)

    Both sugarcane (Saccharum officinarum) and sweet sorghum (Sorghum bicolor) crops are members of the grass (Poaceae) family, and consist of stalks rich in soluble sugars. The extracted juice from both of these crops contains insoluble starch, with much greater quantities occurring in sweet sorghum. ...

  2. Preparation of Proper Immunogen by Cloning and Stable Expression of cDNA coding for Human Hematopoietic Stem Cell Marker CD34 in NIH-3T3 Mouse Fibroblast Cell Line

    Science.gov (United States)

    Shafaghat, Farzaneh; Abbasi-Kenarsari, Hajar; Majidi, Jafar; Movassaghpour, Ali Akbar; Shanehbandi, Dariush; Kazemi, Tohid

    2015-01-01

    Purpose: Transmembrane CD34 glycoprotein is the most important marker for identification, isolation and enumeration of hematopoietic stem cells (HSCs). We aimed in this study to clone the cDNA coding for human CD34 from KG1a cell line and stably express in mouse fibroblast cell line NIH-3T3. Such artificial cell line could be useful as proper immunogen for production of mouse monoclonal antibodies. Methods: CD34 cDNA was cloned from KG1a cell line after total RNA extraction and cDNA synthesis. Pfu DNA polymerase-amplified specific band was ligated to pGEMT-easy TA-cloning vector and sub-cloned in pCMV6-Neo expression vector. After transfection of NIH-3T3 cells using 3 μg of recombinant construct and 6 μl of JetPEI transfection reagent, stable expression was obtained by selection of cells by G418 antibiotic and confirmed by surface flow cytometry. Results: 1158 bp specific band was aligned completely to reference sequence in NCBI database corresponding to long isoform of human CD34. Transient and stable expression of human CD34 on transfected NIH-3T3 mouse fibroblast cells was achieved (25% and 95%, respectively) as shown by flow cytometry. Conclusion: Cloning and stable expression of human CD34 cDNA was successfully performed and validated by standard flow cytometric analysis. Due to murine origin of NIH-3T3 cell line, CD34-expressing NIH-3T3 cells could be useful as immunogen in production of diagnostic monoclonal antibodies against human CD34. This approach could bypass the need for purification of recombinant proteins produced in eukaryotic expression systems. PMID:25789221

  3. Cloning, analysis and functional annotation of expressed sequence tags from the Earthworm Eisenia fetida

    Science.gov (United States)

    Pirooznia, Mehdi; Gong, Ping; Guan, Xin; Inouye, Laura S; Yang, Kuan; Perkins, Edward J; Deng, Youping

    2007-01-01

    Background Eisenia fetida, commonly known as red wiggler or compost worm, belongs to the Lumbricidae family of the Annelida phylum. Little is known about its genome sequence although it has been extensively used as a test organism in terrestrial ecotoxicology. In order to understand its gene expression response to environmental contaminants, we cloned 4032 cDNAs or expressed sequence tags (ESTs) from two E. fetida libraries enriched with genes responsive to ten ordnance related compounds using suppressive subtractive hybridization-PCR. Results A total of 3144 good quality ESTs (GenBank dbEST accession number EH669363–EH672369 and EL515444–EL515580) were obtained from the raw clone sequences after cleaning. Clustering analysis yielded 2231 unique sequences including 448 contigs (from 1361 ESTs) and 1783 singletons. Comparative genomic analysis showed that 743 or 33% of the unique sequences shared high similarity with existing genes in the GenBank nr database. Provisional function annotation assigned 830 Gene Ontology terms to 517 unique sequences based on their homology with the annotated genomes of four model organisms Drosophila melanogaster, Mus musculus, Saccharomyces cerevisiae, and Caenorhabditis elegans. Seven percent of the unique sequences were further mapped to 99 Kyoto Encyclopedia of Genes and Genomes pathways based on their matching Enzyme Commission numbers. All the information is stored and retrievable at a highly performed, web-based and user-friendly relational database called EST model database or ESTMD version 2. Conclusion The ESTMD containing the sequence and annotation information of 4032 E. fetida ESTs is publicly accessible at . PMID:18047730

  4. The role of phenylpropanoid pathway metabolites in resistance of sorghum to pathogens

    Science.gov (United States)

    Sorghum is being developed for diverse uses, including for bioenergy and food. In order to increase efficiency of ethanol production from plant materials, sorghum lines with reduced lignin were developed by incorporating two mutations in lignin biosynthesis pathway genes: brown midrib (bmr) 6 and bm...

  5. Cloning, Expression, and Chromosomal Stabilization of the Propionibacterium shermanii Proline Iminopeptidase Gene (pip) for Food-Grade Application in Lactococcus lactis

    NARCIS (Netherlands)

    Leenhouts, Kees; Bolhuis, Albert; Boot, Johan; Deutz, Inge; Toonen, Marjolein; Venema, Gerard; Kok, Jan; Ledeboer, Aat

    1998-01-01

    Proline iminopeptidase produced by Propionibacterium shermanii plays an essential role in the flavor development of Swiss-type cheeses. The enzyme (Pip) was purified and characterized, and the gene (pip) was cloned and expressed in Escherichia coli and Lactococcus lactis, the latter species being an

  6. Using Genotyping by Sequencing to Map Two Novel Anthracnose Resistance Loci in Sorghum bicolor.

    Science.gov (United States)

    J Felderhoff, Terry; M McIntyre, Lauren; Saballos, Ana; Vermerris, Wilfred

    2016-07-07

    Colletotrichum sublineola is an aggressive fungal pathogen that causes anthracnose in sorghum [Sorghum bicolor (L.) Moench]. The obvious symptoms of anthracnose are leaf blight and stem rot. Sorghum, the fifth most widely grown cereal crop in the world, can be highly susceptible to the disease, most notably in hot and humid environments. In the southeastern United States the acreage of sorghum has been increasing steadily in recent years, spurred by growing interest in producing biofuels, bio-based products, and animal feed. Resistance to anthracnose is, therefore, of paramount importance for successful sorghum production in this region. To identify anthracnose resistance loci present in the highly resistant cultivar 'Bk7', a biparental mapping population of F3:4 and F4:5 sorghum lines was generated by crossing 'Bk7' with the susceptible inbred 'Early Hegari-Sart'. Lines were phenotyped in three environments and in two different years following natural infection. The population was genotyped by sequencing. Following a stringent custom filtering protocol, totals of 5186 and 2759 informative SNP markers were identified in the two populations. Segregation data and association analysis identified resistance loci on chromosomes 7 and 9, with the resistance alleles derived from 'Bk7'. Both loci contain multiple classes of defense-related genes based on sequence similarity and gene ontologies. Genetic analysis following an independent selection experiment of lines derived from a cross between 'Bk7' and sweet sorghum 'Mer81-4' narrowed the resistance locus on chromosome 9 substantially, validating this QTL. As observed in other species, sorghum appears to have regions of clustered resistance genes. Further characterization of these regions will facilitate the development of novel germplasm with resistance to anthracnose and other diseases. Copyright © 2016 Felderhoff et al.

  7. Granivorous birds and sorghum crop in the province of Villa Clara,Cuba

    Directory of Open Access Journals (Sweden)

    Orlando Miguel Saucedo Castillo

    2017-07-01

    Full Text Available In order to reduce the damages granivorous birds cause to sorghum (Sorghum bicolor L. Moench in the province of Villa Clara, Cuba, research based on the determination of the major endemic, migratory birds and their relationship with the distribution were made space of historical meteorological variables in the province in the seasonal behavior of birds in different climatic regions. Population to sorghum producers grouped in different forms surveys were conducted, which yielded a large database, such as the determination of the main grain-eating birds percentage damage incurred, varieties, grain color, growth stage and other indicators. Nine main species affecting sorghum grain-eating birds in our province were recorded; Passer domesticus, Lonchura malacca, Lonchura punctulata, Dives atroviolaceus, Passerina cyanea, Zonotrichia leucophrys, Columbina passerine, Zenaida macroura y Zenaida asiatica. The spatial distribution of meteorological variables and their relation to the seasonal behavior of birds in different climatic regions of the province was determined, based on record four preferential habitat areas. The results allowed us to provide companies and different forms of production in Villa Clara, the possibility of a varietal structure planting of sorghum on the basis of different preferential areas granivorous birds, together with the morphological and physiological characteristics of different genotypes introduced in agricultural production of the province and nationally.

  8. The murine ufo receptor: molecular cloning, chromosomal localization and in situ expression analysis.

    Science.gov (United States)

    Faust, M; Ebensperger, C; Schulz, A S; Schleithoff, L; Hameister, H; Bartram, C R; Janssen, J W

    1992-07-01

    We have cloned the mouse homologue of the ufo oncogene. It encodes a novel tyrosine kinase receptor characterized by a unique extracellular domain containing two immunoglobulin-like and two fibronectin type III repeats. Comparison of the predicted ufo amino acid sequences of mouse and man revealed an overall identity of 87.6%. The ufo locus maps to mouse chromosome 7A3-B1 and thereby extends the known conserved linkage group between mouse chromosome 7 and human chromosome 19. RNA in situ hybridization analysis established the onset of specific ufo expression in the late embryogenesis at day 12.5 post coitum (p.c.) and localized ufo transcription to distinct substructures of a broad spectrum of developing tissues (e.g. subepidermal cells of the skin, mesenchymal cells of the periosteum). In adult animals ufo is expressed in cells forming organ capsules as well as in connective tissue structures. ufo may function as a signal transducer between specific cell types of mesodermal origin.

  9. Fuel ethanol production from sweet sorghum using repeated-batch fermentation.

    Science.gov (United States)

    Chohnan, Shigeru; Nakane, Megumi; Rahman, M Habibur; Nitta, Youji; Yoshiura, Takanori; Ohta, Hiroyuki; Kurusu, Yasurou

    2011-04-01

    Ethanol was efficiently produced from three varieties of sweet sorghum using repeated-batch fermentation without pasteurization or acidification. Saccharomyces cerevisiae cells could be recycled in 16 cycles of the fermentation process with good ethanol yields. This technique would make it possible to use a broader range of sweet sorghum varieties for ethanol production. Copyright © 2010 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  10. Map-based cloning and expression analysis of BMR-6 in sorghum

    Indian Academy of Sciences (India)

    The samples were then ground in a mill and passed .... Figure 2. The histological sections of midrib between WT (a and c) and bmr-6 (b and d). Journal of Genetics ... Segregation for green and brown midrib in F2 populations from two crosses.

  11. Cloning of the human androgen receptor cDNA

    International Nuclear Information System (INIS)

    Govindan, M.V.; Burelle, M.; Cantin, C.; Kabrie, C.; Labrie, F.; Lachance, Y.; Leblanc, G.; Lefebvre, C.; Patel, P.; Simard, J.

    1988-01-01

    The authors discuss how in order to define the functional domains of the human androgen receptor, complementary DNA (cDNA) clones encoding the human androgen receptor (hAR) have been isolated from a human testis λgtll cDNA library using synthetic oligonnucleotide probes, homologous to segments of the human glucocorticoid, estradiol and progesterone receptors. The cDNA clones corresponding to the human glucocorticoid, estradiol and progesterone receptors were eliminated after cross-hybridization with their respective cDNA probes and/or after restriction mapping of the cDNA clones. The remaining cDNA clones were classified into different groups after analysis by restriction digestion and cross-hybridization. Two of the largest cDNA clones from each group were inserted into an expression vector in both orientations. The linearized plasmids were used as templates in in vitro transcription with T7 RNA polymerase. Subsequent in vitro translation of the purified transcripts in rabbit reticulocyte lysate followed by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) permitted the characterization of the encoded polyeptides. The expressed proteins larger than 30,000 Da were analyzed for their ability to bind tritium-labelled dihydrotestosterone ([ 3 H] DHT) with high affinity and specificity

  12. Molecular cloning of the Coch gene of guinea pig inner ear and its expression analysis in cultured fibrocytes of the spiral ligament.

    Science.gov (United States)

    Li, Lishu; Ikezono, Tetsuo; Sekine, Kuwon; Shindo, Susumu; Matsumura, Tomohiro; Pawankar, Ruby; Ichimiya, Issei; Yagi, Toshiaki

    2010-08-01

    We have cloned guinea pig Coch cDNA and the sequence information will be useful for future molecular study combined with physiological experiments. Proper Coch gene expression appears to be dependent on the unique extracellular micro-environment of the inner ear in vivo. These results provide insight into the Coch gene expression and its regulation. To characterize the guinea pig Coch gene, we performed molecular cloning and expression analysis in the inner ear and cultured fibrocytes of the spiral ligament. The Coch cDNA was isolated using RACE. Cochlin isofoms were studied by Western blot using three different types of mammalian inner ear. The cochlear fibrocytes were cultured and characterized by immunostaining. Coch gene expression in the fibrocytes was investigated and the influence of cytokine stimulation was evaluated. The full-length 1991 bp Coch cDNA that encodes a 553 amino acid protein was isolated. The sequence had significant homology with other mammals, and the sizes of the Cochlin isoforms were identical. In the cultured fibrocytes, Coch mRNA was expressed in a very small amount and the isoform production was different, compared with the results in vivo. Cytokine stimulation did not alter the level of mRNA expression or isoform formation.

  13. Expansion of the gateway multisite recombination cloning toolkit.

    Science.gov (United States)

    Shearin, Harold K; Dvarishkis, Alisa R; Kozeluh, Craig D; Stowers, R Steven

    2013-01-01

    Precise manipulation of transgene expression in genetic model organisms has led to advances in understanding fundamental mechanisms of development, physiology, and genetic disease. Transgene construction is, however, a precondition of transgene expression, and often limits the rate of experimental progress. Here we report an expansion of the modular Gateway MultiSite recombination-cloning platform for high efficiency transgene assembly. The expansion includes two additional destination vectors and entry clones for the LexA binary transcription system, among others. These new tools enhance the expression levels possible with Gateway MultiSite generated transgenes and make possible the generation of LexA drivers and reporters with Gateway MultiSite cloning. In vivo data from transgenic Drosophila functionally validating each novel component are presented and include neuronal LexA drivers, LexAop2 red and green fluorescent synaptic vesicle reporters, TDC2 and TRH LexA, GAL4, and QF drivers, and LexAop2, UAS, and QUAS channelrhodopsin2 T159C reporters.

  14. Household production of sorghum beer in Benin: technological and socio-economic aspects

    NARCIS (Netherlands)

    Kayodé, A.P.P.; Hounhouigan, J.D.; Nout, M.J.R.; Niehof, A.

    2007-01-01

    This study evaluated the sorghum brewing microenterprises in Benin with emphasis on the beer quality, the social significance of the product as well as the income generated. Tchoukoutou, the Benin opaque sorghum beer, has important social functions as it fosters the cooperative spirit and remains an

  15. Supplemental irrigation for grain sorghum production in the US Eastern Coastal Plain

    Science.gov (United States)

    Grain sorghum is an important grain crop throughout the world and is generally considered drought tolerant. Recently, in the US eastern Coastal Plain region, there was an emphasis on increasing regional grain production with grain sorghum having an important role. The region soils have low water hol...

  16. Intercropping Urochloa brizantha and sorghum inoculated with Azospirillum brasilense for silage

    Directory of Open Access Journals (Sweden)

    Allan Hisashi Nakao

    Full Text Available ABSTRACT Livestock performance in the Brazilian Cerrado has been limited by the low availability of good quality fodder, especially during periods of low rainfall. The aim of this study was to evaluate growth and dry matter production in two cultivars of sorghum, inoculated or not with diazotrophic bacteria, and as a monocrop or intercropped with palisade grass under a system of crop-livestock integration. The experiments were carried out in the field in the Cerrado region during the autumn-winter period of 2015 and 2016, on the experimental farm of the Faculty of Engineering at Ilha Solteira, UNESP, in Selvíria, in the State of Mato Groso do Sul, Brazil (MS. A randomised complete block experimental design was used in a 2 x 2 x 2 factorial scheme with four replications. The treatments corresponded to two agricultural years (2015 and 2016; the cultivation of dual-purpose grain sorghum, alone or intercropped with palisade grass; with or without inoculation of the sorghum seeds with the bacterium Azospirillum brasilense. The dry matter production of the plant components and plant growth were evaluated for the preparation of silage. Inoculation of sorghum seeds with the bacterium Azospirillum brasilense increases the production of plant dry matter for silage, irrespective of the cultivar or intercrop. Dual-purpose grain sorghum intercropped with palisade grass is a viable agronomic system for producing plant matter for silage during the autumn season.

  17. Cloning and expression of SgCYP450-4 from Siraitia grosvenorii

    Directory of Open Access Journals (Sweden)

    Dongping Tu

    2016-10-01

    Full Text Available CYP450 plays an essential role in the development and growth of the fruits of Siraitia grosvenorii. However, little is known about the SgCYP450-4 gene in S. grosvenorii. Here, based on transcriptome data, a full-length cDNA sequence of SgCYP450-4 was cloned by reverse transcriptase-polymerase chain reaction (RT-PCR and rapid-amplification of cDNA ends (RACE strategies. SgCYP450-4 is 1677 bp in length (GenBank accession No. AEM42985.1 and contains a complete open reading frame (ORF of 1422 bp. The deduced protein was composed of 473 amino acids, the molecular weight is 54.01 kDa, the theoretical isoelectric point (PI is 8.8, and the protein was predicted to possess cytochrome P450 domains. SgCYP450-4 gene was highly expressed in root, diploid fruit and fruit treated with hormone and pollination. At 10 days after treatment with pollination and hormones, the expression of SgCYP450-4 had the highest level and then decreased over time, which was consistent with the development of fruits of S. Grosvenorii. Hormonal treatment could significantly induce the expression of SgCYP450-4. These results provide a reference for regulation of fruit development and the use of parthenocarpy to generate seedless fruit, and provide a scientific basis for the production of growth regulator application agents.

  18. Animal cloning: problems and prospects.

    Science.gov (United States)

    Wells, D N

    2005-04-01

    An efficient animal cloning technology would provide many new opportunities for livestock agriculture, human medicine, and animal conservation. Nuclear cloning involves the production of animals that are genetically identical to the donor cells used in a technique known as nuclear transfer (NT). However, at present it is an inefficient process: in cattle, only around 6% of the embryos transferred to the reproductive tracts of recipient cows result in healthy, longterm surviving clones. Of concern are the high losses throughout gestation, during birth and in the post-natal period through to adulthood. Many of the pregnancy losses relate to failure of the placenta to develop and function correctly. Placental dysfunction may also have an adverse influence on postnatal health. These anomalies are probably due to incorrect epigenetic reprogramming of the donor genome following NT, leading to inappropriate patterns of gene expression during the development of clones. Whilst some physiological tests on surviving clones suggest normality, other reports indicate a variety of post-natal clone-associated abnormalities. This variability in outcome may reflect species-specific and/or cloning methodological differences. Importantly, to date it appears that these clone-associated phenotypes are not transmitted to offspring following sexual reproduction. This indicates that they represent epigenetic errors, rather than genetic errors, which are corrected during gametogenesis. Whilst this needs confirmation at the molecular level, it provides initial confidence in the first application of NT in agriculture, namely, the production of small numbers of cloned sires from genetically elite bulls, for natural mating, to effectively disseminate genetic gain. In addition to the animal welfare concerns with the technology, the underlying health of the animals and the consequential effect on food safety are critical aspects that require investigation to gain regulatory and consumer

  19. The Effect of Vesicular Arbuscular Mycorrhizal (VAM on Yield and Yield Components of Three Sorghum (Sorghum bicolor Cultivars

    Directory of Open Access Journals (Sweden)

    A. Mehraban

    2012-10-01

    Full Text Available To evaluate the influence of vesicular arbuscular mycorrhizal (VAM on yield and yield components of three sorghum cultivars, a factorial experiment based randomized complete block design with four replications was carried out in 2007, at the Agricultural Research Center of Zahak, Iran. The treatments were different mycorrhiza species in three levels: without mycorrhiza (M1, Glomus etanicatum (M2 and G. mosseae(M3 and three cultivars of sorghum: local cultivars (C1, KGS25 (C2 and KGS29 (C3. The results showed that all of the traits measured were increased by inoculation of cultivars with mycorrhiza. The highest plant height (165.1 cm, stem diameter (1.61 cm, flag leaf length (27.22 cm, flag leaf width (3.67 cm and ear width (5.00 cm was obtained by inoculation of seed with Glumus etanicatum, and highest ear length (19.21 cm, ear number (2.51, seed number per ear (10252.11, 1000-seed weight (17.56 g and grain yield (1967.32 kg/ha by using Glumus mossea. The highest leaf width and length belonged to local cultivar, and the highest seed yield to KGS 29 cultivar. However, differences of other traits among sorghum cultivars were not significant. Based on the experimental results it can be concluded that highest grain yield may be obtained by inoculating seeds of KGS 29 with Glumus mossea.

  20. Molecular cloning and gene expression analysis of Ercc6l in Sika deer (Cervus nippon hortulorum.

    Directory of Open Access Journals (Sweden)

    Yupeng Yin

    Full Text Available BACKGROUND: One important protein family that functions in nucleotide excision repair (NER factors is the SNF2 family. A newly identified mouse ERCC6-like gene, Ercc6l (excision repair cross-complementing rodent repair deficiency, complementation group 6-like, has been shown to be another developmentally related member of the SNF2 family. METHODOLOGY/PRINCIPAL FINDINGS: In this study, Sika deer Ercc6l cDNA was first cloned and then sequenced. The full-length cDNA of the Sika deer Ercc6l gene is 4197 bp and contains a 3732 bp open reading frame that encodes a putative protein of 1243 amino acids. The similarity of Sika deer Ercc6l to Bos taurus Ercc6l is 94.05% at the amino acid sequence level. The similarity, however, is reduced to 68.42-82.21% when compared to Ercc6l orthologs in other mammals and to less than 50% compared to orthologs in Gallus gallus and Xenopus. Additionally, the expression of Ercc6l mRNA was investigated in the organs of fetal and adult Sika deer (FSD and ASD, respectively by quantitative RT-PCR. The common expression level of Ercc6l mRNA in the heart, liver, spleen, lung, kidney, and stomach from six different developmental stages of 18 Sika deer were examined, though the expression levels in each organ varied among individual Sika deer. During development, there was a slight trend toward decreased Ercc61 mRNA expression. The highest Ercc6l expression levels were seen at 3 months old in every organ and showed the highest level of detection in the spleen of FSD. The lowest Ercc6l expression levels were seen at 3 years old. CONCLUSIONS/SIGNIFICANCE: We are the first to successfully clone Sika deer Ercc6l mRNA. Ercc6l transcript is present in almost every organ. During Sika deer development, there is a slight trend toward decreased Ercc61 mRNA expression. It is possible that Ercc6l has other roles in embryonic development and in maintaining the growth of animals.

  1. Fuel ethanol production from sweet sorghum bagasse using microwave irradiation

    International Nuclear Information System (INIS)

    Marx, Sanette; Ndaba, Busiswa; Chiyanzu, Idan; Schabort, Corneels

    2014-01-01

    Sweet sorghum is a hardy crop that can be grown on marginal land and can provide both food and energy in an integrated food and energy system. Lignocellulose rich sweet sorghum bagasse (solid left over after starch and juice extraction) can be converted to bioethanol using a variety of technologies. The largest barrier to commercial production of fuel ethanol from lignocellulosic material remains the high processing costs associated with enzymatic hydrolysis and the use of acids and bases in the pretreatment step. In this paper, sweet sorghum bagasse was pretreated and hydrolysed in a single step using microwave irradiation. A total sugar yield of 820 g kg −1 was obtained in a 50 g kg −1 sulphuric acid solution in water, with a power input of 43.2 kJ g −1 of dry biomass (i.e. 20 min at 180 W power setting). An ethanol yield based on total sugar of 480 g kg −1 was obtained after 24 h of fermentation using a mixed culture of organisms. These results show the potential for producing as much as 0.252 m 3  tonne −1 or 33 m 3  ha −1 ethanol using only the lignocellulose part of the stalks, which is high enough to make the process economically attractive. - Highlights: • Different sweet sorghum cultivars were harvested at 3 and 6 months. • Sweet sorghum bagasse was converted to ethanol. • Microwave pretreatment and hydrolysis was done in a single step. • Sugar rich hydrolysates were converted to ethanol using co-fermentation

  2. Cloning, expression and location of RNase9 in human epididymis

    Directory of Open Access Journals (Sweden)

    Lin YQ

    2008-11-01

    Full Text Available Abstract Background Mammalian spermatozoa become fully motile and fertile during transit through the luminal fluid of the epididymis. At least 200 proteins are present in the epididymal lumen, but the potential roles of these luminal proteins in male fertility are unknown. Investigation of the function of these proteins will elucidate the mechanism of sperm maturation, and also provide new drug targets for male contraception. We cloned RNase9 from a human epididymis cDNA library for characterization and analysis of its functions. Findings It was predicted that human RNase9 gene was located on chromosome 14q11.2 and encoded a 205 amino acids protein with a signal peptide of 26 amino acids at the N-terminus. The protein had eight conserved cysteine residues characteristic of the RNase A family members and several potential post-translational modification sites. At the transcriptional level, RNase9 was expressed in a wide variety of tissues, and the expression was higher in men than in boys. RNase9 was localized to the post-equatorial region of the sperms' head. Immunofluorescence staining showed that RNase9 protein was present mostly in the epithelium of the epididymal tubule. Recombinant RNase9 had no ribonuclease activity. In addition, RNase9 had no detectable effect on sperm motility and fertilization as demonstrated by blocking spermatozoa with anti-RNase9 polyclonal serum. Conclusion RNase9 is expressed in a wide variety of tissues. It is located on the post-equatorial region of the sperm head and the epithelium of epididymal tubule. Although RNase9 belongs to the RNase A family, it has no ribonuclease activity.

  3. Cloning, Expression, and Characterization of a Novel Thermophilic Monofunctional Catalase from Geobacillus sp. CHB1.

    Science.gov (United States)

    Jia, Xianbo; Chen, Jichen; Lin, Chenqiang; Lin, Xinjian

    2016-01-01

    Catalases are widely used in many scientific areas. A catalase gene (Kat) from Geobacillus sp. CHB1 encoding a monofunctional catalase was cloned and recombinant expressed in Escherichia coli (E. coli), which was the first time to clone and express this type of catalase of genus Geobacillus strains as far as we know. This Kat gene was 1,467 bp in length and encoded a catalase with 488 amino acid residuals, which is only 81% similar to the previously studied Bacillus sp. catalase in terms of amino acid sequence. Recombinant catalase was highly soluble in E. coli and made up 30% of the total E. coli protein. Fermentation broth of the recombinant E. coli showed a high catalase activity level up to 35,831 U/mL which was only lower than recombinant Bacillus sp. WSHDZ-01 among the reported catalase production strains. The purified recombinant catalase had a specific activity of 40,526 U/mg and K m of 51.1 mM. The optimal reaction temperature of this recombinant enzyme was 60°C to 70°C, and it exhibited high activity over a wide range of reaction temperatures, ranging from 10°C to 90°C. The enzyme retained 94.7% of its residual activity after incubation at 60°C for 1 hour. High yield and excellent thermophilic properties are valuable features for this catalase in industrial applications.

  4. Cloning, Expression, and Characterization of a Novel Thermophilic Monofunctional Catalase from Geobacillus sp. CHB1

    Science.gov (United States)

    2016-01-01

    Catalases are widely used in many scientific areas. A catalase gene (Kat) from Geobacillus sp. CHB1 encoding a monofunctional catalase was cloned and recombinant expressed in Escherichia coli (E. coli), which was the first time to clone and express this type of catalase of genus Geobacillus strains as far as we know. This Kat gene was 1,467 bp in length and encoded a catalase with 488 amino acid residuals, which is only 81% similar to the previously studied Bacillus sp. catalase in terms of amino acid sequence. Recombinant catalase was highly soluble in E. coli and made up 30% of the total E. coli protein. Fermentation broth of the recombinant E. coli showed a high catalase activity level up to 35,831 U/mL which was only lower than recombinant Bacillus sp. WSHDZ-01 among the reported catalase production strains. The purified recombinant catalase had a specific activity of 40,526 U/mg and K m of 51.1 mM. The optimal reaction temperature of this recombinant enzyme was 60°C to 70°C, and it exhibited high activity over a wide range of reaction temperatures, ranging from 10°C to 90°C. The enzyme retained 94.7% of its residual activity after incubation at 60°C for 1 hour. High yield and excellent thermophilic properties are valuable features for this catalase in industrial applications. PMID:27579320

  5. YIELD AND QUALITY OF SORGHUM IN IRRIGATED AGRO LANDSCAPES OF REPUBLIC OF DAGESTAN

    Directory of Open Access Journals (Sweden)

    M G. Muslimov

    2016-01-01

    Full Text Available Aim. One of drought-resistant crops that can provide stable high yields is sorghum, which is salt-tolerant, heat-resistant and a flexible crop of versatile use (green forage, silage, hay, grass meal, grain forage. The research conducted in 2010-2013 included studies on the effectiveness of the methods and norms of sowing the sorghum, required quantities of mineral fertilizers to increase the crop yields and nutritional value of sorghum sown in the irrigated lowland areas of Dagestan. Methods. We conducted three field researches. In experiments with grain sorghum (the middle ripening group Zernogradskiy 88 we studied drill and broad-cast methods of sowing, seeding rate, the calculated doses of mineral fertilizers on programmable levels of crop yields: 6 t/ha (N160P112K70, 7 t/ha (N190P128K80 and 8 t/ha (N220P144K90. Seeding rate was 300, 350 and 400 thousand viable seeds per 1 ha; broadcast was chosen as a sowing method.A field experiment with sweet sorghum included promising hybrid crop Debut, fertilizers N140P80K70, N190P110K95 and N240P140K120 to obtain 60, 70 and 80 t/ha of green mass for two mowings, respectively. Results. The use of fertilizers based on a given level of productivity at optimum plant population can significantly improve the nutritional regime of the soil during the growing season of the sweet sorghum and create optimal conditions for nitrogen, phosphorus and potassium security for the crops and thus obtain the planned crop yield. Conclusion. The fodder quality of sweet sorghum varies depending on the nutrient status of the soil and mowing time.

  6. Cloning, expression, purification, crystallization and preliminary X-ray crystallographic analysis of bacterioferritin A from Mycobacterium tuberculosis

    International Nuclear Information System (INIS)

    Gupta, Vibha; Gupta, Rakesh K.; Khare, Garima; Salunke, Dinakar M.; Tyagi, Anil K.

    2008-01-01

    The cloning, purification and crystallization of a bacterioferritin from M. tuberculosis together with preliminary X-ray characterization of its crystals are reported. Bacterioferritins (Bfrs) comprise a subfamily of the ferritin superfamily of proteins that play an important role in bacterial iron storage and homeostasis. Bacterioferritins differ from ferritins in that they have additional noncovalently bound haem groups. To assess the physiological role of this subfamily of ferritins, a greater understanding of the structural details of bacterioferritins from various sources is required. The gene encoding bacterioferritin A (BfrA) from Mycobacterium tuberculosis was cloned and expressed in Escherichia coli. The recombinant protein product was purified by affinity chromatography on a Strep-Tactin column and crystallized with sodium chloride as a precipitant at pH 8.0 using the vapour-diffusion technique. The crystals diffracted to 2.1 Å resolution and belonged to space group P4 2 , with unit-cell parameters a = 123.0, b = 123.0, c = 174.6 Å

  7. Identification of genetic markers linked to anthracnose resistance in sorghum using association analysis.

    Science.gov (United States)

    Upadhyaya, Hari D; Wang, Yi-Hong; Sharma, Rajan; Sharma, Shivali

    2013-06-01

    Anthracnose in sorghum caused by Colletotrichum sublineolum is one of the most destructive diseases affecting sorghum production under warm and humid conditions. Markers and genes linked to resistance to the disease are important for plant breeding. Using 14,739 SNP markers, we have mapped eight loci linked to resistance in sorghum through association analysis of a sorghum mini-core collection consisting of 242 diverse accessions evaluated for anthracnose resistance for 2 years in the field. The mini-core was representative of the International Crops Research Institute for the Semi-Arid Tropics' world-wide sorghum landrace collection. Eight marker loci were associated with anthracnose resistance in both years. Except locus 8, disease resistance-related genes were found in all loci based on their physical distance from linked SNP markers. These include two NB-ARC class of R genes on chromosome 10 that were partially homologous to the rice blast resistance gene Pib, two hypersensitive response-related genes: autophagy-related protein 3 on chromosome 1 and 4 harpin-induced 1 (Hin1) homologs on chromosome 8, a RAV transcription factor that is also part of R gene pathway, an oxysterol-binding protein that functions in the non-specific host resistance, and homologs of menthone:neomenthol reductase (MNR) that catalyzes a menthone reduction to produce the antimicrobial neomenthol. These genes and markers may be developed into molecular tools for genetic improvement of anthracnose resistance in sorghum.

  8. [Cloning, subcellular localization, and heterologous expression of ApNAC1 gene from Andrographis paniculata].

    Science.gov (United States)

    Wang, Jian; Qi, Meng-Die; Guo, Juan; Shen, Ye; Lin, Hui-Xin; Huang, Lu-Qi

    2017-03-01

    Andrographis paniculata is widely used as medicinal herb in China for a long time and andrographolide is its main medicinal constituent. To investigate the underlying andrographolide biosynthesis mechanisms, RNA-seq for A. paniculata leaves with MeJA treatment was performed. In A. paniculata transcriptomic data, the expression pattern of one member of NAC transcription factor family (ApNAC1) matched with andrographolide accumulation. The coding sequence of ApNAC1 was cloned by RT-PCR, and GenBank accession number was KY196416. The analysis of bioinformatics showed that the gene encodes a peptide of 323 amino acids, with a predicted relative molecular weight of 35.9 kDa and isoelectric point of 6.14. To confirm the subcellular localization, ApNAC1-GFP was transiently expressed in A. paniculata protoplast. The results indicated that ApNAC1 is a nucleus-localized protein. The analysis of real-time quantitative PCR revealed that ApNAC1 gene predominantly expresses in leaves. Compared with control sample, its expression abundance sharply increased with methyl jasmonate treatment. Based on its expression pattern, ApNAC1 gene might involve in andrographolide biosynthesis. ApNAC1 was heterologously expressed in Escherichia coli and recombinant protein was purified by Ni-NTA agarose. Further study will help us to understand the function of ApNAC1 in andrographolide biosynthesis. Copyright© by the Chinese Pharmaceutical Association.

  9. Molecular cloning and expression analysis of a zebrafish novel zinc finger protein gene rnf141

    Directory of Open Access Journals (Sweden)

    Wenqian Deng

    2009-01-01

    Full Text Available ZNF230 is a novel zinc finger gene cloned by our laboratory. In order to understand the potential functions of this gene in vertebrate development, we cloned the zebrafish orthologue of human ZNF230, named rnf141. The cDNA fragment of rnf141 was obtained by rapid amplification of cDNA ends (RACE. The open reading frame (ORF encodes a polypeptide of 222 amino acids which shares 75.65% identity with the human ZNF230. RT-PCR analysis in zebrafish embryo and adult tissues revealed that rnf141 transcripts are maternally derived and that rnf141 mRNA has a broad distribution. Zygotic rnf141 message is strongly localized in the central nervous system, as shown by whole-mount in situ hybridization. Knockdown and over expression of rnf141 can induce abnormal phenotypes, including abnormal development of brain, as well as yolk sac and axis extendsion. Marker gene analysis showed that rnf141 may play a role in normal dorsoventral patterning of zebrafish embryos, suggesting that rnf141 may have a broad function during early development of vertebrates.

  10. Characterization of novel Brown midrib 6 mutations affecting lignin biosynthesis in sorghum

    Science.gov (United States)

    The presence of lignin reduces the quality of lignocellulosic biomass for forage materials and feedstock for biofuels. In C4 grasses, the brown midrib phenotype has been linked to mutations to genes in the monolignol biosynthesis pathway. For example, the Bmr6 gene in sorghum (Sorghum bicolor) has b...

  11. Critical periods of sorghum and palisadegrass in intercropped cultivation for climatic risk zoning

    Directory of Open Access Journals (Sweden)

    Nino Rodrigo Cabral de Barros Lima

    2011-07-01

    Full Text Available The objective of this work was to define critical periods for sorghum and palisadegrass cultivated on crop-livestock integrated systems under water deficit. An experiment was carried out in a completely random block design with four treatments (control and interruption of water supply in three periods and three replicates. Water supply was interrupted until soil water humidity was close to permanent wilting point at the phases: germination of palisadegrass seeds; start of tillering of palisadegrass and initiation of panicles of shorghum; start of shorghum flowering. Water deficit starting at palisadegrass germination delayed intital development of the plants because of the reduction in tillering. Water restriction at panicle initiation phase and at sorghum flowering determined reduction of grain production. Critical periods for intercrop of sorghum and palisadegrass correspond to palisadegrass germination phase and flowering and panicle inititation phase of sorghum.

  12. Evidence for an evolutionarily conserved interaction between cell wall biosynthesis and flowering in maize and sorghum

    Directory of Open Access Journals (Sweden)

    Thompson Karen J

    2002-01-01

    Full Text Available Abstract Background Factors that affect flowering vary among different plant species, and in the grasses in particular the exact mechanism behind this transition is not fully understood. The brown midrib (bm mutants of maize (Zea mays L., which have altered cell wall composition, have different flowering dynamics compared to their wild-type counterparts. This is indicative of a link between cell wall biogenesis and flowering. In order to test whether this relationship also exists in other grasses, the flowering dynamics in sorghum (Sorghum bicolor (L. Moench were investigated. Sorghum is evolutionarily closely related to maize, and a set of brown midrib (bmr mutants similar to the maize bm mutants is available, making sorghum a suitable choice for study in this context. Results We compared the flowering time (time to half-bloom of several different bmr sorghum lines and their wild-type counterparts. This revealed that the relationship between cell wall composition and flowering was conserved in sorghum. Specifically, the mutant bmr7 flowered significantly earlier than the corresponding wild-type control, whereas the mutants bmr2, bmr4, bmr6, bmr12, and bmr19 flowered later than their wild-type controls. Conclusion The change in flowering dynamics in several of the brown midrib sorghum lines provides evidence for an evolutionarily conserved mechanism that links cell wall biosynthesis to flowering dynamics. The availability of the sorghum bmr mutants expands the germplasm available to investigate this relationship in further detail.

  13. Molecular markers associated with aluminium tolerance in Sorghum bicolor.

    Science.gov (United States)

    Too, Emily Jepkosgei; Onkware, Augustino Osoro; Were, Beatrice Ang'iyo; Gudu, Samuel; Carlsson, Anders; Geleta, Mulatu

    2018-01-01

    Sorghum ( Sorghum bicolor , L. Moench) production in many agro-ecologies is constrained by a variety of stresses, including high levels of aluminium (Al) commonly found in acid soils. Therefore, for such soils, growing Al tolerant cultivars is imperative for high productivity. In this study, molecular markers associated with Al tolerance were identified using a mapping population developed by crossing two contrasting genotypes for this trait. Four SSR ( Xtxp34 , Sb5_236 , Sb6_34 , and Sb6_342 ), one STS ( CTG29_3b ) and three ISSR ( 811_1400 , 835_200 and 884_200 ) markers produced alleles that showed significant association with Al tolerance. CTG29_3b, 811_1400 , Xtxp34 and Sb5_ 236 are located on chromosome 3 with the first two markers located close to Alt SB , a locus that underlie the Al tolerance gene ( SbMATE ) implying that their association with Al tolerance is due to their linkage to this gene. Although CTG29_3b and 811_ 1400 are located closer to Alt SB , Xtxp34 and Sb5_236 explained higher phenotypic variance of Al tolerance indices. Markers 835_200 , 884_200 , Sb6_34 and Sb6_342 are located on different chromosomes, which implies the presence of several genes involved in Al tolerance in addition to S bMATE in sorghum. These molecular markers have a high potential for use in breeding for Al tolerance in sorghum.

  14. Cloning, sequencing, and expression of interferon-γ from elk in North America

    Science.gov (United States)

    Sweeney, Steven J.; Emerson, Carlene; Eriks, Inge S.

    2001-01-01

    Eradication of Mycobacterium bovis relies on accurate detection of infected animals, including potential domestic and wildlife reservoirs. Available diagnostic tests lack the sensitivity and specificity necessary for accurate detection, particularly in infected wildlife populations. Recently, an in vitro diagnostic test for cattle which measures plasma interferon-gamma (IFN-γ) levels in blood following in vitro incubation with M. bovis purified protein derivative has been enveloped. This test appears to have increased sensitivity over traditional testing. Unfortunately, it does not detect IFN-γ from Cervidae. To begin to address this problem, the IFN-γ gene from elk (Cervus elaphus) was cloned, sequenced, expressed, and characterized. cDNA was cloned from mitogen stimulated peripheral blood mononuclear cells. The predicted amino acid (aa) sequence was compared to known sequences from cattle, sheep, goats, red deer (Cervus elaphus), humans, and mice. Biological activity of the recombinant elk IFN-γ (rElkIFN-γ) was confirmed in a vesicular stomatitis virus cytopathic effect reduction assay. Production of monoclonal antibodies to IFN-γ epitopes conserved between ruminant species could provide an important tool for the development of reliable, practical diagnostic assays for detection of a delayed type hypersensitivity response to a variety of persistent infectious agents in ruminants, including M. bovis and Brucella abortus. Moreover, development of these reagents will aid investigators in studies to explore immunological responses of elk that are associated with resistance to infectious diseases.

  15. Genetic diversity among sorghum landraces and polymorphism ...

    African Journals Online (AJOL)

    breeding program through marker-assisted selection. ... Keywords: Sorghum, diversity, stay-green trait, marker, polymorphism. ..... Na: Number of different alleles; Na Freq: Frequency of different alleles; Ne: Number of effective alleles; ...

  16. Yield and forage value of a dual-purpose bmr-12 sorghum hybrid

    Science.gov (United States)

    Grain sorghum [Sorghum bicolor (L.) Moench] is an important crop for rainfed production systems with 2.7 million ha grown in the USA in 2013. The brown-midrib (bmr) mutations, especially bmr-12, have resulted in low stover lignin and high fiber digestibility without reducing grain yield in some sor...

  17. Genetic analysis of male sterility genes in different A and B sorghum ...

    African Journals Online (AJOL)

    Hybrid seed production requires use of cytoplasmic male sterility (CMS). Without this system, hybrid seed production would not be economically feasible. There is, therefore, need for developing A and B sorghum lines, as an essential step for development of hybrid sorghum industry. A genetic study of male sterility in ...

  18. Morphophysiological characteristic analysis demonstrated the potential of sweet sorghum (Sorghum bicolor (L.) Moench) in the phytoremediation of cadmium-contaminated soils.

    Science.gov (United States)

    Jia, Weitao; Lv, Sulian; Feng, Juanjuan; Li, Jihong; Li, Yinxin; Li, Shizhong

    2016-09-01

    Cadmium (Cd) contamination is a worldwide environmental problem, and remediation of Cd pollution is of great significance for food production as well as human health. Here, the responses of sweet sorghum cv. 'M-81E' to cadmium stress were studied for its potential as an energy plant in restoring soils contaminated by cadmium. In hydroponic experiments, the biomass of 'M-81E' showed no obvious change under 10 μM cadmium treatment. Cadmium concentration was the highest in roots of seedlings as well as mature plants, but in agricultural practice, the valuable and harvested parts of sweet sorghum are shoots, so promoting the translocation of cadmium to shoots is of great importance in order to improve its phytoremediation capacity. Further histochemical assays with dithizone staining revealed that cadmium was mainly concentrated in the stele of roots and scattered in intercellular space of caulicles. Moreover, the correlation analysis showed that Cd had a negative relationship with iron (Fe), zinc (Zn), and manganese (Mn) in caulicles and leaves and a positive relationship with Fe in roots. These results implied that cadmium might compete with Fe, Zn, and Mn for the transport binding sites and further prevent their translocation to shoots. In addition, transmission electron microscopic observations showed that under 100 μM cadmium treatment, the structure of chloroplast was impaired and the cell wall of vascular bundle cells in leaves and xylem and phloem cells in roots turned thicker compared to control. In summary, morphophysiological characteristic analysis demonstrated sweet sorghum can absorb cadmium and the growth is not negatively affected by mild level cadmium stress; thus, it is a promising material for the phytoremediation of cadmium-contaminated soils considering its economic benefit. This study also points out potential strategies to improve the phytoremediation capacity of sweet sorghum through genetic modification of transporters and cell wall

  19. The ura5 gene of the ascomycete Sordaria macrospora: molecular cloning, characterization and expression in Escherichia coli.

    Science.gov (United States)

    Le Chevanton, L; Leblon, G

    1989-04-15

    We cloned the ura5 gene coding for the orotate phosphoribosyl transferase from the ascomycete Sordaria macrospora by heterologous probing of a Sordaria genomic DNA library with the corresponding Podospora anserina sequence. The Sordaria gene was expressed in an Escherichia coli pyrE mutant strain defective for the same enzyme, and expression was shown to be promoted by plasmid sequences. The nucleotide sequence of the 1246-bp DNA fragment encompassing the region of homology with the Podospora gene has been determined. This sequence contains an open reading frame of 699 nucleotides. The deduced amino acid sequence shows 72% similarity with the corresponding Podospora protein.

  20. Simultaneous inclusion of sorghum and cottonseed meal or millet in broiler diets: effects on performance and nutrient digestibility.

    Science.gov (United States)

    Batonon-Alavo, D I; Bastianelli, D; Lescoat, P; Weber, G M; Umar Faruk, M

    2016-07-01

    Two experiments were conducted to investigate the use of sorghum, cottonseed meal and millet in broiler diets and their interaction when they are used simultaneously. In Experiment 1, a corn-soybean meal control diet was compared with eight experimental treatments based on low tannin sorghum (S30, S45 and S60), cottonseed meal (CM15, CM40) or both ingredients included in the same diet (S30/CM40, S45/CM25 and S60CM15). Results showed that BW gain was not affected by the inclusion of sorghum or cottonseed meal. However, feed intake tended to be affected by the cereal type with the highest values with sorghum-based diets. Feed conversion ratio increased (Pdigestibility (%) of protein and energy with the cottonseed meal and sorghum/cottonseed meal-based diets having lower protein and energy digestibility compared with corn-based diets. In Experiment 2, a control diet was compared with six diets in which corn was substituted at 60%, 80% or 100% by either sorghum or millet and other three diets with simultaneous inclusion of these two ingredients (S30/M30, S40/M40, S50/M50). Single or combined inclusion of sorghum and millet resulted in similar feed intake and growth performance as the control diet. Apparent ileal digestibility of protein and energy was higher with millet-based diets (Pdigestibility of protein in sorghum and millet-based diets tended to decrease linearly with the increasing level of substitution. Sorghum-based diets resulted in lower total tract digestibility of fat compared with millet and sorghum/millet-based diets (Pdigestibility of starch were obtained with the control diet and millet-based diets compared with the sorghum-based treatments. Results of the two experiments suggest that broiler growth performance was not affected by the dietary level of sorghum, millet or cottonseed meal. Nutrient digestion can, however, be affected by these feed ingredients.

  1. Compartmentation of sucrose during radial transfer in mature sorghum culm

    Directory of Open Access Journals (Sweden)

    Vietor Donald M

    2007-06-01

    Full Text Available Abstract Background The sucrose that accumulates in the culm of sorghum (Sorghum bicolor (L. Moench and other large tropical andropogonoid grasses can be of commercial value, and can buffer assimilate supply during development. Previous study conducted with intact plants showed that sucrose can be radially transferred to the intracellular compartment of mature ripening sorghum internode without being hydrolysed. In this study, culm-infused radiolabelled sucrose was traced between cellular compartments and among related metabolites to determine if the compartmental path of sucrose during radial transfer in culm tissue was symplasmic or included an apoplasmic step. This transfer path was evaluated for elongating and ripening culm tissue of intact plants of two semidwarf grain sorghums. The metabolic path in elongating internode tissue was also evaluated. Results On the day after culm infusion of the tracer sucrose, the specific radioactivity of sucrose recovered from the intracellular compartment of growing axillary-branch tissue was greater (nearly twice than that in the free space, indicating that sucrose was preferentially transferred through symplasmic routes. In contrast, the sucrose specific radioactivity in the intracellular compartment of the mature (ripening culm tissue was probably less (about 3/4's than that in free space indicating that sucrose was preferentially transferred through routes that included an apoplasmic step. In growing internodes of the axillary branch of sorghum, the tritium label initially provided in the fructose moiety of sucrose molecules was largely (81% recovered in the fructose moiety, indicating that a large portion of sucrose molecules is not hydrolysed and resynthesized during radial transfer. Conclusion During radial transfer of sucrose in ripening internodes of intact sorghum plants, much of the sucrose is transferred intact (without hydrolysis and resynthesis and primarily through a path that includes an

  2. [Cloning and expression of Micrococcus luteus IAM 14879 Rpf and its role in the recovery of the VBNC state in Rhodococcus sp. DS471].

    Science.gov (United States)

    Ding, Linxian; Zhang, Pinghua; Hong, Huachang; Lin, Hongjun; Yokota, Akira

    2012-01-01

    The purpose of the present study was to produce the Rpf (resuscitation promoting factor) protein by cloning and expressing the rpf gene, secreted by Micrococcus luteus IAM 14879, in Escherichia coli and to evaluate its role in the recovery of the VBNC (viable but non-culturable) state in high-GC Gram-positive bacteria. Genomic DNA was extracted from Micrococcus luteus IAM 14879 and the rpf gene was amplified by PCR using specific primers. The PCR products was purified, cloned into a pET15b expression vector, and transformed into Escherichia coli BL21 (DE3). Then the pET15b plasmid expression vector was used to confirm the purification of the recombinant proteins via SDS-PAGE. The VBNC state cells from the high-GC Gram-positive bacteria, Rhodococcus sp. DS471, were used to confirm the promotion and recovery of growth capacity. Rhodococcus sp. DS471 were isolated from soil and closely related to Micrococcus luteus IAM 14879. The gene sequences confirmed that the rpf gene from Micrococcus luteus IAM 14879 that was expressed in Escherichia coli, was 672 bp. SDS-PAGE analysis showed that the recombinant Rpf protein was obtained successfully, and further studies showed it capable of promoting the recovery of the VBNC state by about 100-fold relative to the control. Rpf of Micrococus luteus IAM 14879 can be successfully cloned and expressed in Escherichia coli and shows a strong ability to promote the recovery of the VBNC state of cells of Rhodococcus sp. DS471.

  3. [Clone, construct, expression and verification of lactoferricin B gene and several sequence mutations in yeast].

    Science.gov (United States)

    Feng, Yong-qian; Zha, Xiao-jun; Zhai, Chao-yang

    2007-07-01

    To construct the eucaryotic recombinant plasmid of pYES2/LactoferricinB expressing in yeast of S. cerevisiae, of which the expressed protein antibacterial activity was verified in preliminary. By self-template PCR method, the gene of Lactoferricin B and its several sequence mutations were amplified with the parts of the pre-synthesized single chains. And then Lactoferricin B gene and its mutants were cloned into the vector of pYES2 to construct the recombined expression plasmid pYES2/Lactoferricin B etc. extracted and used to transform the yeast S. cerevisiae. The expressions of proteins were determined after induced by galactose. The expression proteins were collected and purified by hydronium-exchange column, and the bacterial inhibited test was applied to identify the protein antibacterial activities. The PCR amplifying and DNA sequencing tests indicated that the purpose plasmid contained the Lactoferricin B gene and several mutations. The induced target proteins were confirmed by SDS-PAGE electrophoresis and mass spectrum test. The protein antibacterial activities of mutations were verified in preliminary. The recombined plasmid pYES2/Lactoferricin B etc. are successfully constructed and induced to express in yeast cell of S. cerevisiae; the obtained recombined protein of Lactoferricin B provides a basis for further research work on the biological function and antibacterial activity.

  4. Dicty_cDB: SSM365 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available WGS-SbicolorF (JM107 adapted methyl filtered) Sorghum bicolor genomic clone hv90e08 5', DNA sequence. 36 1.2... 2 BZ330175 |BZ330175.1 hv90e08.g1 WGS-SbicolorF (JM107 adapted methyl filtered) ...rF (JM107 adapted methyl filtered) Sorghum bicolor genomic clone hz26f02 5', DNA sequence. 36 1.4 2 BZ628823... |BZ628823.1 ih62d12.g1 WGS-SbicolorF (DH5a methyl filtered) Sorghum bicolor geno...mic clone ih62d12 5', DNA sequence. 36 1.7 2 BZ340606 |BZ340606.1 ic39h07.b1 WGS-SbicolorF (JM107 adapted methyl filter

  5. High-polyphenol sorghum bran extract inhibits cancer cell growth through DNA damage, cell cycle arrest, and apoptosis

    Science.gov (United States)

    As diet is one of the major controllable factors in cancer development, potentially chemopreventive foods are of significant interest to public health. One such food is sorghum (Sorghum bicolor), a cereal grain that contains varying concentrations of polyphenols. In a panel of 15 sorghum germplasm...

  6. Characterization of novel multi-seeded (msd) mutants of sorghum for increasing grain number

    Science.gov (United States)

    The tribe Andropogoneae of the Poaceae family exhibits highly branched inflorescence known as panicle or tassel. Characteristically, each spikelet in a panicle or tassel comprise of a combination of sessile/fertile and sterile florets. In sorghum, (Sorghum bicolor L. Moench), the existing cultivars ...

  7. Factors affecting the porridge-making quality in South African sorghums

    CSIR Research Space (South Africa)

    Taylor, JRN

    1997-04-01

    Full Text Available fermented, sour porridges remain popular, particularly among the Tswana of Botswana and South Africa (Novellie 1982; Sooliman 1993) The production of sorghum porridge involves ?rst producing a meal from sorghum grain. Commercially, this is generally done..., South Africa. the remaining part of the kernel (essentially endosperm) into a coarse meal. Alternatively, endosperm meal can be produced directly from grain by roller milling (Munck 1995). The meal is then cooked with boiling water into a porridge...

  8. Using elevated CO{sub 2} to increase the biomass of a Sorghum vulgare x Sorghum vulgare var. sudanense hybrid and Trifolium pratense L. and to trigger hyperaccumulation of cesium

    Energy Technology Data Exchange (ETDEWEB)

    Wu Huibin [Centre for Research in Ecotoxicology and Environmental Remediation, Institute of Agro-Environmental Protection, Ministry of Agriculture, Tianjin 300191 (China); Open Key Laboratory of Agro-environment and Agro-product Safety of the Ministry of Agriculture, Tianjin (China); College of Resources and Environment, Huazhong Agricultural University, 430070 Wuhan, Hubei Province (China); Tang Shirong, E-mail: tangshir@hotmail.com [Centre for Research in Ecotoxicology and Environmental Remediation, Institute of Agro-Environmental Protection, The Ministry of Agriculture, Tianjin 300191 (China); Open Key Laboratory of Agro-environment and Agro-product Safety of the Ministry of Agriculture, Tianjin (China); Zhang Ximei; Guo Junkang; Song, Zhengguo; Tian Shuai [Centre for Research in Ecotoxicology and Environmental Remediation, Institute of Agro-Environmental Protection, Ministry of Agriculture, Tianjin 300191 (China); Open Key Laboratory of Agro-environment and Agro-product Safety of the Ministry of Agriculture, Tianjin (China); Smith, Donald L. [Plant Science Department, McGill University, Macdonald Campus, 21111 Lakeshore Road, Ste. Anne de Bellevue, Quebec, H9X 3V9 (Canada)

    2009-10-30

    The most important challenge to use phytoremediation is how to improve its efficiency by increasing the accumulation of metals in plants, or by improving key plant biological traits that should enhance metal uptake. In this paper, we used open-top chambers to investigate the effects of elevated CO{sub 2} (860 {mu}L L{sup -1}) on biomass and Cs uptake by a Sorghum vulgare x Sorghum vulgare var. sudanense hybrid and Trifolium pratense L. growing on soils spiked with various levels of cesium (0, 300, 1500 and 3000 mg Cs kg{sup -1}). The results showed that elevated CO{sub 2} not only increased aboveground biomass of the Sorghum and Trifolium species by 32-111%, and by 8-11%, respectively, compared to the ambient CO{sub 2} treatment, but also caused more accumulation of Cs by Sorghum species (up to 73%) than Trifolium species (up to 43%). It was speculated that the increase in biomass and the improvement in Cs accumulation ability at elevated CO{sub 2} could be related to lowered soil pH values, and changes in number and kind of microorganisms in the rhizospheres of the two tested species. This is the first report of a link among elevated CO{sub 2}, increased biomass and hyperaccumulation of Cs by Sorghum and Trifolium species.

  9. Variation of Transpiration Efficiency in Sorghum

    Science.gov (United States)

    Declining freshwater resources, increasing population, and growing demand for biofuels pose new challenges for agriculture research. To meet these challenges, the concept “Blue Revolution” was proposed to improve water productivity in agriculture--“More Crop per Drop”. Sorghum is the fifth most imp...

  10. Variation in transpiration efficiency in sorghum

    Science.gov (United States)

    Declining freshwater resources, increasing population, and growing demand for biofuels pose new challenges for agriculture research. To meet these challenges, the concept “Blue Revolution” was proposed to improve water productivity in agriculture--“More Crop per Drop”. Sorghum is the fifth most imp...

  11. On the extent of genetic variation for transpiration efficiency in sorghum

    International Nuclear Information System (INIS)

    Hammer, G.L.; Broad, I.J.; Farquhar, G.D.

    1997-01-01

    A glasshouse study examined 49 diverse sorghum lines for variation in transpiration efficiency. Three of the 49 lines grown were Sorghum spp. native to Australia; one was the major weed Johnson grass (Sorghum halepense), and the remaining 45 lines were cultivars of Sorghum bicolor. All plants were grown under non-limiting water and nutrient conditions using a semi-automatic pot watering system designed to facilitate accurate measurement of water use. Plants were harvested 56-58 days after sowing and dry weights of plant parts were determined. Transpiration efficiency differed significantly among cultivars. The 3 Australian native sorghums had much lower transpiration efficiency than the other 46 cultivars, which ranged from 7.7 to 6.0 g/kg. For the 46 diverse cultivars, the ratio of range in transpiration efficiency to its l.s.d. was 2.0, which was similar to that found among more adapted cultivars in a previous study. This is a significant finding as it suggests that there is likely to be little pay-off from pursuing screening of unadapted material for increased variation in transpiration efficiency. It is necessary, however, also to examine absolute levels of transpiration efficiency to determine whether increased levels have been found. The cultivar with greatest transpiration efficiency in this study (IS9710) had a value 9% greater (P < 0.05) than the accepted standard for adapted sorghum cultivars. The potential impact of such an increase in transpiration efficiency warrants continued effort to capture it. Transpiration efficiency has been related theoretically and experimentally to the degree of carbon isotope discrimination in leaf tissue in sorghum, which thus offers a relatively simple selection index. In this study, the variation in transpiration efficiency was not related simply to carbon isotope discrimination. Significant associations of transpiration efficiency with ash content and indices of photosynthetic capacity were found. However, the

  12. Comparative potentials of native arbuscular mycorrhizal fungi to improve nutrient uptake and biomass of Sorghum bicolor Linn

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    Pattarawadee Sumthong Nakmee

    2016-05-01

    Full Text Available Sorghum (Sorghum bicolor Linn. seedlings were grown in pots using Pakchong soil from Nakhon Ratchasima province. Ten species of native Arbuscular mycorrhizal (AM fungi: Glomus sp. 1, Glomus sp. 2, Glomus sp. 3, Glomus aggregatum, Glomus fasciculatum, Acaulospora longula, Glomus occultum, Acaulospora scrobiculata, Acaulospora spinosa and Scutellospora sp., were used to inoculate sorghum seedlings. The sorghum growth and uptake of several major nutrients were evaluated at the harvesting stage. The results revealed that sorghum inoculated with A. scrobiculata produced the greatest biomass, grain dry weight and total nitrogen uptake in shoots. The highest phosphorus uptake in shoots was found in A. spinosa-inoculated plants, followed by Glomus sp. and A. scrobiculata, whereas Scutellospora sp.-inoculated plants showed the highest potassium uptake in shoots followed by A. scrobiculata. Overall, the most efficient AM fungi for improvement of nutrient uptake, biomass and grain dry weight in sorghum were A. scrobiculata.

  13. Exploitation of sweet sorghum biomass for biofuel production using mixed acidogenic and methanogenic cultures and pure cultures of ruminococcus albus

    International Nuclear Information System (INIS)

    Ntaikou, I.; Antonopoulou, G.; Marazioti, C.; Lyberatos, G.

    2008-01-01

    Full text: The present study focuses on the exploitation of sweet sorghum biomass for gas biofuel production in continuous and batch systems. Sweet sorghum is an annual C 4 plant of tropical origin, well-adapted to sub-tropical and temperate regions and highly productive in biomass. It is rich in readily fermentable sugars and thus it can be considered as an excellent raw material for biohydrogen production from many different fermentative microorganisms. Extraction of free sugars from the sorghum stalks was achieved using water at 30 degrees centigrade. After the extraction process a liquid fraction (sorghum extract), rich in sucrose, and a solid fraction (sorghum cellulosic-hemicellulosic residues or sorghum bagasse), containing the cellulose and hemicelluloses, were obtained. A two-step continuous process was developed for the biological hydrogen production and the subsequent production of biogas from sweet sorghum extract. In the first reactor sugars were fermented to hydrogen, volatile fatty acids and alcohols b mixed acidogenic culture derived from the indigenous microfauna of sweet sorghum. The hydrogen producing reactor was operated at five different hydraulic retention times (HRT), i.e 24h, 12h, 8h, 6h and 4h. The HRT of 12h proved to be the most effective leading to the production 10.4 L H 2 /kg sweet sorghum biomass. Subsequently, the effluent was fed to the methanogenic reactor, where all the residual organic compounds were digested by an acclimated methanogenic culture derived from activated sludge. The operation of the methanogenic reactor was studied at three different HRTs, i.e 20d, 15d and 10d with the latter being the most prosing leading to the production 35.2 L CH 4 /kg sweet sorghum biomass. Both continuous and batch cultures were used for the investigation of hydrogen production from sweet sorghum biomass using Ruminococcus albus. R. albus is an important, fibrolytic bacterium of the rumen that can hydrolyse both cellulose and hemicellulose

  14. Response of Fusarium thapsinum to sorghum brown midrib lines and to phenolic metabolites

    Science.gov (United States)

    Presentation type: poster Presentation title: Response of Fusarium thapsinum to sorghum brown midrib lines and to phenolic metabolites Sorghum lines were bred for reduced lignin for cellulosic bioenergy uses, through the incorporation of brown midrib (bmr) bmr6 and/or 12 into two gen...

  15. Genetic diversity of sweet sorghum germplasm in Mexico using AFLP and SSR markers

    Science.gov (United States)

    The objective of this work was to evaluate the diversity and genetic relationships between lines and varieties of the sweet sorghum (Sorghum bicolor) germplasm bank of the National Institute for Forestry, Agriculture and Livestock Research, Mexico, using AFLP and SSR markers. The molecular markers ...

  16. Comparison of Chemical and Degradability Characteristics of Green Forage and Silage of Sorghums Varieties with Corn Using In vitro

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    A. Hedayatipour

    2012-10-01

    Full Text Available The chemical and fermentative parameters of three fresh forages and silages of sorghum including Sweet, Pegah and Speedfeed varieties were compared with corn using in vitro method, also degradability coefficients of forages and silages were determined by in situ method. Forages were planted in the same condition and harvested in soft dough stage, then ensilaged in four replicates for each time of 30, 60 and 90 days of preservation in mini silos. Buffering capacity in green Sweet sorghum was lower than corn and Speedfeed, and acid detergent fiber and water soluble carbohydrates respectively were significantly highest and lowest in fresh forage of Speedfeed sorghum. In time of 60 days, percent of acid detergent lignin of corn silage was lower than Sweet and Speedfeed sorghum silages; similarly, residual water soluble carbohydrate was lowest in corn silage. The lactate Concentration in corn and Pegah sorghums was higher than Sweet and Speedfeed silages. In corn and Sweet sorghum silages, Contents of acetic acid and ammonium nitrogen were highest and lowest, respectively. In nylon bag experiment, Degradation rate of corn and Pegah sorghum forages were significantly higher than Sweet and Speedfeed sorghums that cause to more effective degradability with passage rate of 0.08 in this forages. Also, the slowly degradation coefficient of corn silage was higher than sorghums silages. In conclusion, Speedfeed sorghum forage is not suitable for making silage in comparison others, and corn silage had more potential of degradability.

  17. Cloning and expression of three thaumatin-like protein genes from Polyporus umbellatus

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    Mengmeng Liu

    2017-05-01

    Full Text Available Genes encoding thaumatin-like protein (TLPs are frequently found in fungal genomes. However, information on TLP genes in Polyporus umbellatus is still limited. In this study, three TLP genes were cloned from P. umbellatus. The full-length coding sequence of PuTLP1, PuTLP2 and PuTLP3 were 768, 759 and 561 bp long, respectively, encoding for 256, 253 and 187 amino acids. Phylogenetic trees showed that P. umbellatus PuTLP1, PuTLP2 and PuTLP3 were clustered with sequences from Gloeophyllum trabeum, Trametes versicolor and Stereum hirsutum, respectively. The expression patterns of the three TLP genes were higher in P. umbellatus with Armillaria mellea infection than in the sclerotia without A. mellea. Furthermore, over-expression of three PuTLPs were carried out in Escherichia coli BL21 (DE3 strain, and high quality proteins were obtained using Ni-NTA resin that can be used for preparation of specific antibodies. These results suggest that PuTLP1, PuTLP2 and PuTLP3 in P. umbellatus may be involved in the defense response to A. mellea infections.

  18. Dexamethasone attenuates grain sorghum dust extract-induced increase in macromolecular efflux in vivo.

    Science.gov (United States)

    Akhter, S R; Ikezaki, H; Gao, X P; Rubinstein, I

    1999-05-01

    The purpose of this study was to determine whether dexamethasone attenuates grain sorghum dust extract-induced increase in macromolecular efflux from the in situ hamster cheek pouch and, if so, whether this response is specific. By using intravital microscopy, we found that an aqueous extract of grain sorghum dust elicited significant, concentration-dependent leaky site formation and increase in clearance of FITC-labeled dextran (FITC-dextran; mol mass, 70 kDa) from the in situ hamster cheek pouch (P grain sorghum dust extract- and substance P-induced increases in macromolecular efflux from the in situ hamster cheek pouch in a specific fashion.

  19. Potensi penggunaan beberapa varietas sorgum manis (Sorghum bicolor (L. Moench sebagai tanaman pakan

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    Mustikoweni Purnomohadi

    2012-02-01

    Full Text Available Sweet sorghum is a versatile crop that can be used as grain crop, sugar alcohol production and even as forage crop. The aim of this study was to evaluate the potential use of sweet sorghum either as grain crop or forage crop. The experiment used four varieties of sweet sorghum: Rio, Cawley, Keller and Wray, which were planted in polybag with six replication using Completely Randomized Design. The result of the research showed that Keller and Wray had longer vegetative growth, and good quality of chemical composition for forage than Rio and Cawley.

  20. Cis-regulatory signatures of orthologous stress-associated bZIP transcription factors from rice, sorghum and Arabidopsis based on phylogenetic footprints

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    Xu Fuyu

    2012-09-01

    Full Text Available Abstract Background The potential contribution of upstream sequence variation to the unique features of orthologous genes is just beginning to be unraveled. A core subset of stress-associated bZIP transcription factors from rice (Oryza sativa formed ten clusters of orthologous groups (COG with genes from the monocot sorghum (Sorghum bicolor and dicot Arabidopsis (Arabidopsis thaliana. The total cis-regulatory information content of each stress-associated COG was examined by phylogenetic footprinting to reveal ortholog-specific, lineage-specific and species-specific conservation patterns. Results The most apparent pattern observed was the occurrence of spatially conserved ‘core modules’ among the COGs but not among paralogs. These core modules are comprised of various combinations of two to four putative transcription factor binding site (TFBS classes associated with either developmental or stress-related functions. Outside the core modules are specific stress (ABA, oxidative, abiotic, biotic or organ-associated signals, which may be functioning as ‘regulatory fine-tuners’ and further define lineage-specific and species-specific cis-regulatory signatures. Orthologous monocot and dicot promoters have distinct TFBS classes involved in disease and oxidative-regulated expression, while the orthologous rice and sorghum promoters have distinct combinations of root-specific signals, a pattern that is not particularly conserved in Arabidopsis. Conclusions Patterns of cis-regulatory conservation imply that each ortholog has distinct signatures, further suggesting that they are potentially unique in a regulatory context despite the presumed conservation of broad biological function during speciation. Based on the observed patterns of conservation, we postulate that core modules are likely primary determinants of basal developmental programming, which may be integrated with and further elaborated by additional intrinsic or extrinsic signals in