| Project/Area Number |
03454556
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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 |
分子遺伝学・分子生理学
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| Research Institution | Institute of Molecular and Cellular Biosciences |
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Principal Investigator |
AYUSAWA Dai Institute of Molecular and Cellular Biosciences, Associate Professor, 分子細胞生物学研究所, 助教授 (00142109)
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| Co-Investigator(Kenkyū-buntansha) |
OISHI Michio Institute of Molecular and Cellular Bioscience, Professor, 分子細胞生物学研究所, 教授 (00126004)
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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 |
¥5,100,000 (Direct Cost: ¥5,100,000)
Fiscal Year 1993: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1992: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1991: ¥2,900,000 (Direct Cost: ¥2,900,000)
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| Keywords | Senescence / Immortalization / Gene / Chromosome 7 / Signal transduction / Gene mapping / L1反復配列 / 相補性群 / 不死化細胞株 / 微小核細胞融合法 / 放射線雑種細胞 / 分裂停止遺伝子 / 老化遺伝子 / 正常線維芽細胞 / 不死化 / RFLP解析 / 微少核細胞融合法 / 不死化遺伝子 / 染色体導入 / 染色体マッピング / 不死化細胞 / アンチセンスcDNAライブラリ- |
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| Research Abstract |
Normal somatic cells in mammals stop dividing in culture after a limited number of cell division. This phenomenon is called cellular senescence and governed by several genes. When these genes are inactivated in a cell, it acquires potential to divide indefinitely and immortalize. Genetic analysis revealed that there exist at least four genetic complementation groups (A-D) in immortal human cell lines. We have found that introduction of human chromosme 7 by microcell-mediated chromosome transfer specifically induces senescence in cell lines belonging to group D.Then we made a panel of radiation hybrids under the backgroud of mouse A9 cells. The hybrids were characterized by FISH analysis and PCR with STS markers to determine how fragments of human chromosme 7 are integrated on mouse A9 genome. Of these hybrids, we selected a clone that contains only 2-3 Mb human DNA and can induce senescence in group D cell lines. We rescued human DNA from this clone and used it as a probe to map the putative senescence gene on human chromosme 7. The mapping data are useful for us to identify the gene.
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