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CHO细胞中的分子重组与DNA双链断裂修复

Molecular recombination and the repair of DNA double-strand breaks in CHO cells.

作者信息

Resnick M A, Moore P D

出版信息

Nucleic Acids Res. 1979 Jul 11;6(9):3145-60. doi: 10.1093/nar/6.9.3145.

Abstract

Molecular recombination and the repair of DNA double-strand breaks (DSB) have been examined in the G-0 and S phase of the cell cycle using a temperature-sensitive CHO cell line to test i) if there are cell cycle restrictions on the repair of DSB's' ii) the extent to which molecular recombination can be induced between either sister chromatids or homologous chromosomes and iii) whether repair of DSB's involves recombination (3). Mitomycin C (1-2 micrograms/ml) or ionizing radiation (50 krad) followed by incubation resulted in molecular recombination (hybrid DNA) in S phase cells. Approximately 0.03 to 0.10% of the molecules (number average molecular weight: 5.6 x 10(6) Daltons after shearing) had hybrid regions for more than 75% of their length. However, no recombination was detected in G-0 cells. Since the repair of DSB was observed in both stages with more than 50% of the breaks repaired in 5 hours, it appears that DSB repair in G-0 cells does not involve recombination between homologous chromosomes. The possibility is not excluded that repair in G-0 cells involves only small regions (less than 4 x 10(6) Daltons).

摘要

利用温度敏感型中国仓鼠卵巢(CHO)细胞系,在细胞周期的G-0期和S期对DNA双链断裂(DSB)的分子重组及修复进行了研究,以检验:i)DSB修复是否存在细胞周期限制;ii)姐妹染色单体或同源染色体之间分子重组可被诱导的程度;iii)DSB修复是否涉及重组(3)。丝裂霉素C(1-2微克/毫升)或电离辐射(50拉德)处理后进行孵育,导致S期细胞出现分子重组(杂交DNA)。约0.03%至0.10%的分子(剪切后数均分子量:5.6×10⁶道尔顿)其杂交区域长度超过其总长度的75%。然而,在G-0期细胞中未检测到重组。由于在两个阶段均观察到DSB的修复,且5小时内超过50%的断裂得以修复,因此G-0期细胞中的DSB修复似乎不涉及同源染色体之间的重组。不排除G-0期细胞中的修复仅涉及小区域(小于4×10⁶道尔顿)的可能性。

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