| Project/Area Number |
08555205
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
生物・生体工学
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
IMANAKA Tadayuki Kyoto Univ.Dept.of Synth.Chem.& Biol.Chem., Grad.School of Engin.Professor, 工学研究科, 教授 (30029219)
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| Co-Investigator(Kenkyū-buntansha) |
EMI Shigenori Director, Tsuruga Bio Research Inst.Toyobo Co., 敦賀バイオ研究所, 所長
TAGAGI Masahiro Osaka Univ., Dept of Biotechnology, Grad School of Engin, Asst.Prof., 大学院・工学研究科, 助教授 (00183434)
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| Project Period (FY) |
1996 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥11,300,000 (Direct Cost: ¥11,300,000)
Fiscal Year 1997: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 1996: ¥8,600,000 (Direct Cost: ¥8,600,000)
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| Keywords | Hyperthermophile / Evolution / Genome / Pyrococcus / RNA polmerase / Molecular Chaperon / Nitrogen Metabolism / Eukaryote |
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| Research Abstract |
The gene encoding the beta subunit of a molecular chaperonin from the hyperthermophilic archaeon Pyrococcus sp.strain KOD1 (cpkB) was cloned, sequenced, and expressed in Escherichia coli. The cpkB geme is composed of 1,641 nucleotides, encoding a protein (546 amino acids) with a molecular mass of 59,140 Da. The enhancing effect of CpkB on enzyme sstability was examined by using Saccharomyces cerevisiae alcohol dehydrogenase (ADH). Purified recombinat CpkB prevents thermal denaturation and enhances thermostability of ADH.CpkB requires ATP for its chaperonin function at a low CpkB concentration ; however, CpkB functions without ATP when present in excess. In vivo chapernoin function for the solubilization of insoluble proteins was also studied by coexpressing CpkB and CobQ (cobryic acid synthase), indication that Cpkb is useful for solubilizing the insoluble proteins in vivo. These results suggest that the beta subnit plays a major role in chaperonin acitivity and is functional without the alpha subunit.
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