| Project/Area Number |
04660122
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
応用微生物学・発酵学
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| Research Institution | HIROSHIMA UNIVERSITY |
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Principal Investigator |
TSUCHIYA Eiko Hiroshima University, Dept. Fermentation technol., Associate Professor, 工学部, 助教授 (90127671)
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| Project Period (FY) |
1992 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1993: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1992: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Saccharomyces cerevisiae / Cell-Division Cycle / G2-Phase Control / DNA-Binding Protein / Protein Kinase / Nuclear Protein / Saccharomyces cevevisiae / プロテインチナーゼ |
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| Research Abstract |
The NPSl gene is a novel cell division cycle (CDC) gene of Saccharomyces cerevisiae enconding a 160 kd nuclear protein essenntial for cell growth. I investigated the functions of NPS1 gene in cell cycle and obtained the following results.
1) I constructed a mutant nps1 gene which has a glutamic acid residue at position 792 instead of lysin residue (Lys792) at a suspected ATP-binding site of protein kinase consennsus motief, and found that Lys792 is indispensable for in vivo function of NPS1 gene product.
2) By investigating the phenotypes of conditional lethal strain which can express NPS1 gene only in galactose medium, I found that nps1 gene works in g2 phase of S.cerevisiae transducing the signal of completion of DNA replication.
3) The NPS1 gene product was found to locate in nuclear saffold of S.cerevisiae and possess DNA-binding activity.
4) I succeeded to produce the Nps1 protein in SF9 cell transformed with recombinant baculovirus carrying NPS1 gene. Large scaled culture of transformed SF9 cells and purification of Nps1 protein are now on going.
5) In order to identify genes acting in relation to NPS1, I carried out the following experiments : i) isolate mutant whose growth is suppressed by the high dosage-expression of NPS1 gene ; ii) screen the high copy suppressor of lethality caused by NPS1 depletion ; iii) screen the gene which cause synthetic lethality with high-dosage expression of NPS1 gene. The analyzes of mutants and genes obtained by the above experiments are now on going.
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