| Project/Area Number |
03640573
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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 |
植物生理学
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| Research Institution | OSAKA CITY UNIVERSITY |
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Principal Investigator |
SHIMODA Chikashi Osaka City University, Faculty of Science, Associate Professor, 理学部, 助教授 (80047290)
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| Project Period (FY) |
1991 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1993: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1992: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1991: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Keywords | Fission yeast / meiosis / sexual reproduction / RNA helicase / mating / specific antibody / DEAD protein / Drosophila / ポリクローナル抗体 / ウエスタンブロット / 細胞周期 / 転写後調節 / PCR法 / 部位特異的変異誘導 / RNAヘリカ-ゼ |
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| Research Abstract |
The fission yeast ste13^+ gene which encodes a putative ATP-dependent RNA helicase is essential for initiation of sexual reproduction in response to nitrogen-starvation signal. The ste13^+ gene and its products were analyzed to verify our hypothesis that Ste13 regulates gene expression during sexual cycle at a post-transcriptional level and the following results were obtained.
(1)In vitro mutagenesis of ste13 :
The ste13 null mutant was defective in mating and meiosis but able to grow in rich medium. Site-directed mutagenesis introduced into the consensus motifs of RNA helicases caused the sterile phenotype. These facts indicate that the ste13 gene is not essential for mitotic cell division but indispensable for induction of sexual cycle.
(2)Intracellular localization of the ste13 protein :
The GST-ste13 fusion proteins were overproduced in E.coli. The affinity-purified fusion protein was used as the antigen to prepare a polyclonal antibody. Western blot analysis revealed that this antibody recognized a 58 kDa polypeptide in S.pombe cell extracts. Immunofluorescence microscopy with anti-ste13 showed that the ste13 protein was not localized in the nucleus and distributed in the cytoplasm.
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