Characterization of X-ray-sensitive cells and isolation of the gene responsible for their radiosensitivity.
| Project/Area Number |
60480504
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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 |
放射線5生物学
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| Research Institution | Faculty of Medicine, University of Tokyo |
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Principal Investigator |
SAKAI Kazuo (1986-1987) Faculty of Medicine, %university of Tokyo, 医学部(医), 助手 (40153837)
岡田 重文 (1985) 東京大学, 医学部, 教授
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| Co-Investigator(Kenkyū-buntansha) |
YASUDA Hideyo Faculty of Pharmacology, Kanazawa University, 薬学部, 助教授 (40111554)
SUZUKI Norio Faculty of Medicine, University of Tokyo, 医学部(医), 教授 (10010050)
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| Project Period (FY) |
1985 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| Budget Amount *help |
¥6,500,000 (Direct Cost: ¥6,500,000)
Fiscal Year 1987: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1986: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1985: ¥4,700,000 (Direct Cost: ¥4,700,000)
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| Keywords | DNA damage / DNA Repair / Radiosensitivity / Cell death / Gene transfer / Ataxia telangiectasia / アタキシア細胞 / AT細胞 / X線感受性 / 融合細胞 |
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| Research Abstract |
The purpose of the present project is to elucidate the mechanism of radiationinduced cell death by characterizing radiosensitive mutant cells and by isolation of a gene responsible for their radiosensitivity.
Toward this goal 4 subjects were set and have been carried out in parallel. I. Isolation of radiosensitive mutants
Cultured cells (human and rodent) were treated with mutagens and selected for radiosensitivity. In mouse L5178Y cells five radiosensitive strains were isolated.
Il. DNA breaks and their repair in radiosensitive cells
In fibroblasts of parients with Ataxia telangiectasia (AT), which were more radiosensitive than normal cells, induction and repair of DNA breaks after X- irradiation were investigated. There was no difference between normal and AT cells either in the number of breaks induced or in the DNA repair profile.
IlI. A new technique which is useful for further characterization of radiosensitive cells
Using a specific DNA probe for DHFR gene, DNA breaks in the gene was analyzed on Southern analysis panels of DNA from X-irradiated cells. DNA breaks was detected as a shift of DHFR signals to lower molecular wight region and/or disappearance of the signal in a dose-dependent manner.
IV. Gene isolation system
Normal human DNA was transfected into UV-and drug-sensitive CHO mutant cells. UV-resistant transformants were picked up and DNA fragments responsible for the transformaiton were identified using human repetitive sequence as a marker. The gene defective in the mutant was found to be the one already isolated by another group, ERCC-1. This results, however, indicates a usefulness of the present strategy for isolation of radiosensitive genes.
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Report
(2 results)
Research Products
(12 results)
