The Experts below are selected from a list of 43275 Experts worldwide ranked by ideXlab platform
Donald L Jarvis - One of the best experts on this subject based on the ideXlab platform.
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Silkworms transformed with chimeric silkworm spider silk genes spin composite silk fibers with improved mechanical properties
Proceedings of the National Academy of Sciences of the United States of America, 2012Co-Authors: Florence Teule, Yun-gen Miao, Malcolm J Fraser, Bonghee Sohn, Joe J Hull, Randolph V Lewis, Donald L JarvisAbstract:The development of a spider silk-manufacturing process is of great interest. However, there are serious problems with natural manufacturing through spider farming, and standard recombinant protein production platforms have provided limited progress due to their inability to assemble spider silk proteins into fibers. Thus, we used piggyBac vectors to create transgenic Silkworms encoding chimeric silkworm/spider silk proteins. The silk fibers produced by these animals were composite materials that included chimeric silkworm/spider silk proteins integrated in an extremely stable manner. Furthermore, these composite fibers were, on average, tougher than the parental silkworm silk fibers and as tough as native dragline spider silk fibers. These results demonstrate that Silkworms can be engineered to manufacture composite silk fibers containing stably integrated spider silk protein sequences, which significantly improve the overall mechanical properties of the parental silkworm silk fibers.
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Silkworms transformed with chimeric silkworm/spider silk genes spin composite silk fibers with improved mechanical properties
Proceedings of the National Academy of Sciences of the United States of America, 2012Co-Authors: Florence Teule, Yun-gen Miao, Malcolm J Fraser, Bonghee Sohn, Joe J Hull, Randolph V Lewis, Young-soo Kim, Donald L JarvisAbstract:The development of a spider silk-manufacturing process is of great interest. However, there are serious problems with natural manufacturing through spider farming, and standard recombinant protein production platforms have provided limited progress due to their inability to assemble spider silk proteins into fibers. Thus, we used piggyBac vectors to create transgenic Silkworms encoding chimeric silkworm/spider silk proteins. The silk fibers produced by these animals were composite materials that included chimeric silkworm/spider silk proteins integrated in an extremely stable manner. Furthermore, these composite fibers were, on average, tougher than the parental silkworm silk fibers and as tough as native dragline spider silk fibers. These results demonstrate that Silkworms can be engineered to manufacture composite silk fibers containing stably integrated spider silk protein sequences, which significantly improve the overall mechanical properties of the parental silkworm silk fibers.
Yun-gen Miao - One of the best experts on this subject based on the ideXlab platform.
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Silkworms transformed with chimeric silkworm spider silk genes spin composite silk fibers with improved mechanical properties
Proceedings of the National Academy of Sciences of the United States of America, 2012Co-Authors: Florence Teule, Yun-gen Miao, Malcolm J Fraser, Bonghee Sohn, Joe J Hull, Randolph V Lewis, Donald L JarvisAbstract:The development of a spider silk-manufacturing process is of great interest. However, there are serious problems with natural manufacturing through spider farming, and standard recombinant protein production platforms have provided limited progress due to their inability to assemble spider silk proteins into fibers. Thus, we used piggyBac vectors to create transgenic Silkworms encoding chimeric silkworm/spider silk proteins. The silk fibers produced by these animals were composite materials that included chimeric silkworm/spider silk proteins integrated in an extremely stable manner. Furthermore, these composite fibers were, on average, tougher than the parental silkworm silk fibers and as tough as native dragline spider silk fibers. These results demonstrate that Silkworms can be engineered to manufacture composite silk fibers containing stably integrated spider silk protein sequences, which significantly improve the overall mechanical properties of the parental silkworm silk fibers.
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Silkworms transformed with chimeric silkworm/spider silk genes spin composite silk fibers with improved mechanical properties
Proceedings of the National Academy of Sciences of the United States of America, 2012Co-Authors: Florence Teule, Yun-gen Miao, Malcolm J Fraser, Bonghee Sohn, Joe J Hull, Randolph V Lewis, Young-soo Kim, Donald L JarvisAbstract:The development of a spider silk-manufacturing process is of great interest. However, there are serious problems with natural manufacturing through spider farming, and standard recombinant protein production platforms have provided limited progress due to their inability to assemble spider silk proteins into fibers. Thus, we used piggyBac vectors to create transgenic Silkworms encoding chimeric silkworm/spider silk proteins. The silk fibers produced by these animals were composite materials that included chimeric silkworm/spider silk proteins integrated in an extremely stable manner. Furthermore, these composite fibers were, on average, tougher than the parental silkworm silk fibers and as tough as native dragline spider silk fibers. These results demonstrate that Silkworms can be engineered to manufacture composite silk fibers containing stably integrated spider silk protein sequences, which significantly improve the overall mechanical properties of the parental silkworm silk fibers.
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cloning and expression of a cellulase gene in the silkworm bombyx mori by improved bac to bac bmnpv baculovirus expression system
Molecular Biology Reports, 2010Co-Authors: Xinghua Li, Roy Bhaskar, Jiabiao Hu, Huajun Yang, Fang Zhou, Dan Wang, Yun-gen MiaoAbstract:Cellulases catalyze the hydrolysis of cellulose which are mainly three types: endoglucanases, cellobiohydrolases and β-glucosidases. It can be used in converting cellulosic biomass to glucose that can be used in different applications such as production of fuel ethanol, animal feed, waste water treatment and in brewing industry. In this paper, we cloned a 1380-bp endoglucanase I (EG I) gene from mycelium of filamentous fungus Trichoderma viride strain AS 3.3711 using PCR-based exon splicing methods, and expressed the recombinant EG I mature peptide protein in both silkworm BmN cell line and silkworm larvae with a newly established Bac-to-Bac/BmNPV mutant baculovirus expression system, which lacks the virus-encoded chitinase (chiA) and cathepsin (v-cath) genes of Bombyx mori nucleopolyhedrovirus (BmNPV). An around 49-kDa protein was visualized after mBacmid/BmNPV/EG I infection, and the maximum expression in silkworm larvae was at 84 h post-infection. The ANOVA showed that the enzymes from recombinant baculoviruses infected Silkworms exhibited significant maximum enzyme activity at the environmental condition of pH 7.0 and temperature 50°C. It was stable at pH range from 5.0 to 10.0 and at temperature range from 50 to 60°C, and increased 24.71 and 22.84% compared with that from wild baculoviruses infected Silkworms and normal Silkworms, respectively. The availability of large quantities of EG I that the silkworm provides maybe greatly facilitate the future research and the potential application in industries.
Florence Teule - One of the best experts on this subject based on the ideXlab platform.
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Silkworms transformed with chimeric silkworm spider silk genes spin composite silk fibers with improved mechanical properties
Proceedings of the National Academy of Sciences of the United States of America, 2012Co-Authors: Florence Teule, Yun-gen Miao, Malcolm J Fraser, Bonghee Sohn, Joe J Hull, Randolph V Lewis, Donald L JarvisAbstract:The development of a spider silk-manufacturing process is of great interest. However, there are serious problems with natural manufacturing through spider farming, and standard recombinant protein production platforms have provided limited progress due to their inability to assemble spider silk proteins into fibers. Thus, we used piggyBac vectors to create transgenic Silkworms encoding chimeric silkworm/spider silk proteins. The silk fibers produced by these animals were composite materials that included chimeric silkworm/spider silk proteins integrated in an extremely stable manner. Furthermore, these composite fibers were, on average, tougher than the parental silkworm silk fibers and as tough as native dragline spider silk fibers. These results demonstrate that Silkworms can be engineered to manufacture composite silk fibers containing stably integrated spider silk protein sequences, which significantly improve the overall mechanical properties of the parental silkworm silk fibers.
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Silkworms transformed with chimeric silkworm/spider silk genes spin composite silk fibers with improved mechanical properties
Proceedings of the National Academy of Sciences of the United States of America, 2012Co-Authors: Florence Teule, Yun-gen Miao, Malcolm J Fraser, Bonghee Sohn, Joe J Hull, Randolph V Lewis, Young-soo Kim, Donald L JarvisAbstract:The development of a spider silk-manufacturing process is of great interest. However, there are serious problems with natural manufacturing through spider farming, and standard recombinant protein production platforms have provided limited progress due to their inability to assemble spider silk proteins into fibers. Thus, we used piggyBac vectors to create transgenic Silkworms encoding chimeric silkworm/spider silk proteins. The silk fibers produced by these animals were composite materials that included chimeric silkworm/spider silk proteins integrated in an extremely stable manner. Furthermore, these composite fibers were, on average, tougher than the parental silkworm silk fibers and as tough as native dragline spider silk fibers. These results demonstrate that Silkworms can be engineered to manufacture composite silk fibers containing stably integrated spider silk protein sequences, which significantly improve the overall mechanical properties of the parental silkworm silk fibers.
Randolph V Lewis - One of the best experts on this subject based on the ideXlab platform.
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Silkworms with Spider Silklike Fibers Using Synthetic Silkworm Chow Containing Calcium Lignosulfonate, Carbon Nanotubes, and Graphene
2019Co-Authors: Xiaoli Zhang, Thomas I. Harris, Ana Laura Licon, Paula F. Oliveira, Blake Taurone, Brittney J. Walsh, Brianne E. Bell, Caleb T. Walker, Bailey J. Mcfarland, Randolph V LewisAbstract:Silkworm silk has become increasingly relevant for material applications. However, the industry as a whole is retracting because of problems with mass production. One of the key problems is the inconsistent properties of the silk. A means by which to improve the silk material properties is through enhanced sericulture techniques. One possible technique is altering the feed of the Silkworms to include single-wall carbon nanotubes (SWNTs) or graphene (GR). Recently published results have demonstrated substantial improvement in fiber mechanical properties. However, the effect of the surfactant used to incorporate those materials into the feed on the fiber mechanical properties in comparison to normal silkworm silk has not been studied or reported. Thus, the total effect of feeding the SWNT and GR in the presence of surfactants on Silkworms is not understood. Our study focuses on the surfactant [calcium lignosulfonate (LGS)] and demonstrates that it alone results in appreciable improvement of mechanical properties in comparison to nontreated silkworm silk. Furthermore, our study demonstrates that mixing the LGS, SWNT, and GR directly into the artificial diet of Silkworms yields improved mechanical properties without decline below the control silk at high doses of SWNT or GR. Combined, we present evidence that mixing surfactants, in this case LGS, directly with the diet of Silkworms creates a high-quality fiber product that can exceed 1 GPa in tensile strength. With the addition of nanocarbons, either SWNT or GR, the improvement is even greater and consistently surpasses control fibers. However, feeding LGS alone is a more economical and practical choice to consistently improve the mechanical properties of silkworm fiber
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Silkworms transformed with chimeric silkworm spider silk genes spin composite silk fibers with improved mechanical properties
Proceedings of the National Academy of Sciences of the United States of America, 2012Co-Authors: Florence Teule, Yun-gen Miao, Malcolm J Fraser, Bonghee Sohn, Joe J Hull, Randolph V Lewis, Donald L JarvisAbstract:The development of a spider silk-manufacturing process is of great interest. However, there are serious problems with natural manufacturing through spider farming, and standard recombinant protein production platforms have provided limited progress due to their inability to assemble spider silk proteins into fibers. Thus, we used piggyBac vectors to create transgenic Silkworms encoding chimeric silkworm/spider silk proteins. The silk fibers produced by these animals were composite materials that included chimeric silkworm/spider silk proteins integrated in an extremely stable manner. Furthermore, these composite fibers were, on average, tougher than the parental silkworm silk fibers and as tough as native dragline spider silk fibers. These results demonstrate that Silkworms can be engineered to manufacture composite silk fibers containing stably integrated spider silk protein sequences, which significantly improve the overall mechanical properties of the parental silkworm silk fibers.
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Silkworms transformed with chimeric silkworm/spider silk genes spin composite silk fibers with improved mechanical properties
Proceedings of the National Academy of Sciences of the United States of America, 2012Co-Authors: Florence Teule, Yun-gen Miao, Malcolm J Fraser, Bonghee Sohn, Joe J Hull, Randolph V Lewis, Young-soo Kim, Donald L JarvisAbstract:The development of a spider silk-manufacturing process is of great interest. However, there are serious problems with natural manufacturing through spider farming, and standard recombinant protein production platforms have provided limited progress due to their inability to assemble spider silk proteins into fibers. Thus, we used piggyBac vectors to create transgenic Silkworms encoding chimeric silkworm/spider silk proteins. The silk fibers produced by these animals were composite materials that included chimeric silkworm/spider silk proteins integrated in an extremely stable manner. Furthermore, these composite fibers were, on average, tougher than the parental silkworm silk fibers and as tough as native dragline spider silk fibers. These results demonstrate that Silkworms can be engineered to manufacture composite silk fibers containing stably integrated spider silk protein sequences, which significantly improve the overall mechanical properties of the parental silkworm silk fibers.
Qingyou Xia - One of the best experts on this subject based on the ideXlab platform.
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Toxicological evaluation of transgenic Silkworms.
Toxicology research, 2020Co-Authors: Liang Jiang, Huizhen Guo, Qiang Sun, Yumei Wang, Enyu Xie, Haoyu Liuli, Qingyou XiaAbstract:Safety of transgenic Silkworms must be evaluated before their commercial application. We assessed subacute toxicity using a 28-day feeding study in rats. Eighty rats were evenly allocated into four groups, with each group containing 10 male and 10 female rats. Rats of three groups were fed dried transgenic silkworm H19.9A pupae with overexpressed endogenous Bmhsp19.9, transgenic silkworm A4SOR pupae with overexpressed exogenous SOR, or normal silkworm pupae at a dose of 3.0 g/kg/day, respectively. The fourth group served as a normal feeding control. The body weight, feed consumption, hematology response variables, serum biochemical parameters, organ weights, gross necropsy, and histopathologic of animals were evaluated. No mortality, adverse effects, or major differences in the evaluated parameters were observed in the groups fed transgenic pupae in comparison with the control, suggesting that transgenic Silkworms are toxicologically equivalent to normal Silkworms and are safe for consumption in rats.
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Enhanced heat tolerance in transgenic silkworm via overexpression of Pyrococcus furiosus superoxide reductase.
Insect biochemistry and molecular biology, 2017Co-Authors: Liang Jiang, Chunlin Huang, Bingbing Wang, Huizhen Guo, Qiang Sun, Fei Xia, Qingyou XiaAbstract:Heat shock causes a serious harm to organisms by accelerating the production of reactive oxygen species (ROS). Pyrococcus furiosus superoxide reductase (PfSOR) is an enzyme that efficiently detoxifies ROS. In order to generate a silkworm strain with high heat tolerance for sericulture, we synthesized an artificial DNA sequence encoding PfSOR based on the codon bias of Bombyx mori. PfSOR was successfully overexpressed in transgenic silkworm (named A4SOR) and BmE cells, as determined by RT-PCR and western blot analyses. An SOR activity assay confirmed that the expressed enzyme was functional in A4SOR. After exposure to a temperature of 35 °C for 44 h, the mortality rate was about 30% lower in transgenic A4SOR than in non-transgenic Silkworms. Moreover, transgene expression had no apparent effect on economic characteristics of Silkworms. The heat tolerance of silkworm was thus enhanced by expressing an archaeal SOR; this can be useful for sericulture in regions where the average temperature exceeds the optimal environmental temperature for B. mori of 25 °C.
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selection of reference genes for analysis of stress responsive genes after challenge with viruses and temperature changes in the silkworm bombyx mori
Molecular Genetics and Genomics, 2016Co-Authors: Huizhen Guo, Liang Jiang, Qingyou XiaAbstract:Viruses and high temperature (HT) are the primary threats to Silkworms. Changes in the expression of stress-response genes can be measured using quantitative polymerase chain reaction (qPCR) after exposure to viruses or HT. However, appropriate reference genes (RGs) for qPCR data normalization have not been established in this organism. In this study, we summarized the RGs used in the previous silkworm studies after infection with Bombyx mori nucleopolyhedrovirus (BmNPV), B. mori cytoplasmic polyhedrosis virus (BmCPV), or B. mori densovirus (BmDNV) or after HT treatment. The expression levels of these RGs were extracted from silkworm transcriptome data to screen for candidate RGs that were unaffected by the experimental conditions. Actin-1 (A1), actin-3 (A3), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and translation initiation factor 4a (TIF-4A) were selected for further qPCR verification. The results of RNA-seq and qPCR showed that GAPDH and TIF-4A were suitable RGs after BmNPV challenge or HT stress, whereas TIF-4A was an appropriate RG for BmCPV or BmDNV-Z challenge in Silkworms. These results suggested that TIF-4A may be the most appropriate RG for gene expression analysis after challenge with viruses or HT in Silkworms.
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the 5 utr intron of the midgut specific bmapn4 gene affects the level and location of expression in transgenic Silkworms
Insect Biochemistry and Molecular Biology, 2015Co-Authors: Liang Jiang, Chunlin Huang, Huizhen Guo, Qiang Sun, Tingcai Cheng, Zhengwen Peng, Yinghui Dang, Weiqiang Liu, Qingyou XiaAbstract:Introns are important for regulating gene expression. BmAPN4, which has a 5'-UTR upstream intron (5 UI), is specifically expressed in the entire silkworm midgut. In our previous study, the promoter region upstream of the 5 UI of BmAPN4 was cloned and identified as the P3 promoter (P3P) with activity only in the anterior midgut. In this study, the sequence consisting of the P3P and the 5 UI was cloned and named as P3P+5 UI. A transgenic vector was constructed in which EGFP was controlled by P3P+5 UI. Transgenic P3+5 UI Silkworms were generated by embryo microinjection. RT-PCR showed P3P+5 UI activity throughout the larval stage. Intense green fluorescence was seen only in the entire midgut of P3+5 UI Silkworms and expression was confirmed by RT-PCR. qPCR revealed that expression of EGFP in the anterior midgut of P3+5 UI Silkworms was 64% higher than in P3 Silkworms, indicating the 5 UI sustained intron-mediated enhancement of gene expression. These results suggested that the BmAPN4 5 UI affected the level and site of expression. The 5 UI was cloned and added behind P2P, another specific promoter with activity only in the anterior midgut of silkworm, to construct the P2P+5 UI and transgenic P2+5 UI Silkworms. Expression patterns were the same for P2P+5 UI and P2P, suggesting that the 5UI of BmAPN4 did not affect P2P. This study found that the BmAPN4 5 UI affected the amount and location of gene expression. Its influence appeared to be dependent on a specific promoter.
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the progress and future of enhancing antiviral capacity by transgenic technology in the silkworm bombyx mori
Insect Biochemistry and Molecular Biology, 2014Co-Authors: Liang Jiang, Qingyou XiaAbstract:Bombyx mori is a common lepidopteran model and an important economic insect for silk production. B. mori nucleopolyhedrovirus (BmNPV) is a typical pathogenic baculovirus that causes serious economic losses in sericulture. B. mori and BmNPV are a model of insect host and pathogen interaction including invasion of the host by the pathogen, host response, and enhancement of host resistance. The antiviral capacity of Silkworms can be improved by transgenic technology such as overexpression of an endogenous or exogenous antiviral gene, RNA interference of the BmNPV gene, or regulation of the immune pathway to inhibit BmNPV at different stages of infection. Antiviral capacity could be further increased by combining different methods. We discuss the future of an antiviral strategy in silkworm, including possible improvement of anti-BmNPV, the feasibility of constructing transgenic Silkworms with resistance to multiple viruses, and the safety of transgenic Silkworms. The silkworm model could provide a reference for disease control in other organisms.
