| Project/Area Number |
08458220
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Molecular biology
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| Research Institution | Osaka University |
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Principal Investigator |
NOJIMA Hiroshi Osaka University, Research Institute for Microbial Diseases, Professor., 微生物病研究所 (30156195)
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| Co-Investigator(Kenkyū-buntansha) |
KIMURA Shinya Osaka University, Research Institute for Microbial Diseases, Technical Official, 微生物病研究所, 教務職員 (70273703)
NABESHIMA Kentaro Osaka University, Research Institute for Microbial Diseases, Research Associate., 微生物病研究所, 助手 (60294120)
TANAKA Seiji Osaka University, Research Institute for Microbial Diseases, Research Associa, 微生物病研究所, 助手 (50263314)
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| Project Period (FY) |
1996 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 1998: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1997: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1996: ¥3,600,000 (Direct Cost: ¥3,600,000)
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| Keywords | Subtraction / cDNA Library / Meiosis / Spermiogenesis / Go phase / Osteoclast / MITF / Metastasis / Gin4 / DNA傷害 / DNA複製開始 / 蛋白質キナーゼ / ヒドロキシ尿素 / 複製ライセンス因子 / サイクリンG / CDK5 / FISH / ウエスタンブロット / モノクローナル抗体 |
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| Research Abstract |
In order to isolate systematically the genes that control cell cycle and cell growth, we have established an improved protocol to prepare a subtracted cDNA library of high quality that allowed the large-scale isolation of transcriptionally induced mRNA (cDNA) species. Using this protocol, we prepared subtracted cDNA libraries of several experimental systems as described below. To perform the efficient and comprehensive isolation of the clones involved in the subtracted cDNA library, we also developed a stepwise subtraction method. By applying these techniques, we have isolated a large number of novel cDNA clones in the following experimental systems. They are comprehensively named as TISP (Transcription increased in spermiogenesis), meu (meiotic expression upregulated), EPOC (Expressed predominantly in osteoclast), TIGA (Transcription increased by growth arrest), ElM (Epxpression increased in mi mouse), TIB (Transcription increased in BL6 mouse), TRIF (Transcription reduced in F2408) and SSR (shear Stress Responsive). By DNA sequencing, we found that about half of these genes are novel, and some of them have not been registered in the gene bank even as EST (expressed sequence tag). We have examined the functions of these genes and found that many of them displayed biologically important functions. These results indicate that our strategy is applicable to a wide variety of biological phenomena in which the trancriptional up or down regulation play the important role in these phenomena and is useful to understand the regulatory mechanisms of otherwise unresolvable biological problems.
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