Translational Enhancement of Recombinant Protein Synthesis in Transgenic Silkworms by a 5'-Untranslated Region of Polyhedrin Gene of Bombyx mori Nucleopolyhedrovirus(GENETICS, MOLECULAR BIOLOGY, AND GENE ENGINEERING)

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概要

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

• 社団法人日本生物工学会の論文
• 2008-06-25

著者

• Yoshizato Katsutoshi

  Yoshizato Morphomatrix Project Erato Jst

• Yoshizato Katsutoshi

  Yoshizato Project Cooperative Link Of Unique Science And Technology For Economy Revitalization Hiros

• Iizuka Masashi

  Yoshizato Project Cooperative Link Of Unique Science And Technology For Economy Revitalization Hiros

• Shimizu Katsuhiko

  Yoshizato Project, Cluster

• Tomita Masahiro

  Yoshizato Project, Cooperative Link of Unique Science and Technology for Economy Revitalization, Hir

• Kikuchi Yutaka

  Developmental Biology Laboratory, Department of Biological Science, Graduate School of Science, Hiro

• Tomita Masahiro

  Yoshizato Project Cooperative Link Of Unique Science And Technology For Economy Revitalization Hiros

• Shimizu Katsuhiko

  Yoshizato Project Cooperative Link Of Unique Science And Technology For Economy Revitalization Hiros

• Shimizu Katsuhiko

  Yoshizato Project Cluster

• Kikuchi Yutaka

  Developmental Biology Laboratory Department Of Biological Science Graduate School Of Science Hiroshi

• Kikuchi Yutaka

  Developmental Biology Laboratory And Hiroshima University 21st Century Coe Program For Advanced Radi

• Yoshizato Katsutoshi

  Yoshizato Morphomatrix Project Erato Jrdc

関連論文

• FUNCTIONS OF A BZIP-CONTAINING TRANSCRIPTION FACTOR DURING XENOPUS LARVAL-TO-ADULT GASTROINTESTINAL REMODELING(Developmental Biology,Abstracts of papers presented at the 75^ Annual Meeting of the Zoological Society of Japan)
• Translational Enhancement of Recombinant Protein Synthesis in Transgenic Silkworms by a 5'-Untranslated Region of Polyhedrin Gene of Bombyx mori Nucleopolyhedrovirus(GENETICS, MOLECULAR BIOLOGY, AND GENE ENGINEERING)
• Metamorphosis-associated and region-specific expression of calbindin gene in the posterior intestinal epithelium of Xenopus laevis larva
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• Editorial
• Amphibian Metamorphosis

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