| Project/Area Number |
01470119
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
応用生物化学・栄養化学
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| Research Institution | Faculty of Agriculture, Ibaraki University |
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Principal Investigator |
SUGAWARA Kiyoshi Ibaraki University, Faculty of Agriculture, Dept of Resource Biology, Professor, 農学部, 教授 (40007662)
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| Co-Investigator(Kenkyū-buntansha) |
FUKAZAWA Chikafusa National Food Res. Inst., Minstry of Agric. Forest. and Fish., Division of Appli, 食品総合研究所・応用微生物部, 室長
TAKAHARA Hidenari Ibarki University, Faculty of Agriculture Dept. of Resonrce Biology, Asit. Profe, 農学部, 助教授 (30122063)
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| Project Period (FY) |
1989 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥7,400,000 (Direct Cost: ¥7,400,000)
Fiscal Year 1990: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1989: ¥5,400,000 (Direct Cost: ¥5,400,000)
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| Keywords | Peptidylarginine deiminase / Protein deiminating enzyme / Primary structure of protein / Isozyme / CDNA cloning / ゲノムDNAクロ-ニング / ペプチジルアルギニンディミナ-ゼ / 蛋白質脱イミノ酸 |
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| Research Abstract |
1). Three types of isozyme of peptidylarginine deiminase were found and its characterization and tissue distribution were described.
2). Molecular cloning and sequencing of cDNA coding for mouse uterus peptidylarginine deiminase were performed. The amini terminus of the enzyme was predicted to be Nーacety1-methionine which corresponds to the initiatione. The MW of the mature enzyme is calculated to be 76,260 from deduced amino acid sequence.
3). The cloning of the genomic DNA was carried out, and the results demonstrated that the mouse peptidylarginine deiminase gene was a single copy gene. Then tow clones which covered 65% of the cDNA were isolated from mouse genomic libraries.
4). Gene expression plasmid of peptidylarginine deiminase cDNA was constracted. The EcoR1 digestsof cDNA fragments, PAD1 and PAD24 were ligated its 5'-EcoR1-3'-HindIII fragment was inserted to E. coil expression vector pKK223-3.
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