| Project/Area Number |
09640805
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
動物生理・代謝
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
KAGEYAMA Takashi Kyoto University, Primate Research Institute, Associate Professor, 霊長類研究所, 助教授 (20027501)
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| Co-Investigator(Kenkyū-buntansha) |
YONEZAWA Satoshi Aichi Human Service Center, Institute for Developmental Research, Leader, 発達障害研究所, 室長 (90001867)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 1998: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1997: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | cathepsin E / protease / aspartic protease / cathepsin E inhibitor / mouse / Ascaris suum / アスパラギン酸プロテアーゼ / 蛋白質分解酵素 / 遺伝子解析 / 回虫インヒビター |
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| Research Abstract |
1. A 16.8-kb genomic clone containing the mouse cathepsin E gene was isolated from a 129/SvJ XFix II library. The gene was determined to be 13,8 kb in length and to consist of 9 exons. Highly conservative sequences were identified in the 5'-flnaking region containing potential binding motifs for GATA-transcriptional factors. The mouse cathepsin E gene was assigned to chromosome 1 band F by fluorescence in situ hybridization.
2. Molecular cloning of a cDNA for a pepsin inhibitor in the round worm, Ascaris suum, was achieved. The ORF was found to encode a 20-residue potential signal peptide and a 149-residue inhibitor moiety. To obtain the active inhibitor, we constracted a yeast expression vector, pYES2API, containing the inducible galactosidase promoter and a DNA fragment containing a fusion protein of the yeast alpha-factor leader and the Ascaris pepsin inhibitor. The active inhibitor was secreted in the culture medium, the yield being approximately 3 mg/I/day, and purified by a two-step procedure that included HPLC.
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