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Genetic diversity of UV-sensitive DNA repair mutants of Chinese hamster ovary cells.

机译:中国仓鼠卵巢细胞对紫外线敏感的DNA修复突变体的遗传多样性

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摘要

Mutant lines of Chinese hamster ovary cells that show hypersensitivity to killing and mutagenesis by UV light were analyzed by genetic complementation analysis to determine whether defects in different gene loci might underlie a common cellular phenotype,. To facilitate rapid screening of mutant clones, a procedure was devised that allowed presumptive complementation to be assessed on the basis of the frequency of UV-resistant cells after fusion by polyethylene glycol. Four classes were identified among 44 clones tested. By using drug-resistance markers for selection of hybrid cells in crosses between UV mutant and wild type, a mutant from each of the four classes was shown to behave as phenotypically recessive. Hybrids were also isolated from crosses between each of the pair combinations of the four mutants. All such hybrids were relatively resistant to UV killing, providing confirmation of the complementation classes. When mutants representing the four UV-complementation classes were tested with the polyaromatic hydrocarbon 7-bromomethylbenz(a)anthracene, complementation was again seen for all pair combinations. These results suggest that each class of mutants represents a biochemical defect that plays a common role in the repair of both UV-induced and chemically induced lesions in the DNA.
机译:通过基因互补分析,分析了对仓鼠卵巢细胞表现出对杀灭和诱变超敏性的突变株,以确定不同基因位点的缺陷是否可能是共同的细胞表型。为了促进突变体克隆的快速筛选,设计了一种程序,该程序允许在聚乙二醇融合后基于抗紫外线细胞的频率评估推定的互补性。在测试的44个克隆中鉴定出四个类别。通过使用抗药性标记在UV突变体和野生型之间的杂交中选择杂交细胞,显示出来自四个类别的突变体表现为表型隐性的。还从四个突变体的每个配对组合之间的杂交中分离了杂种。所有这些杂种均相对抗紫外线杀伤,从而证实了互补类别。当用聚芳烃7-溴甲基苯并(a)蒽测试代表四个UV互补类的突变体时,所有对组合都再次看到互补。这些结果表明,每种类型的突变体都代表一种生化缺陷,该缺陷在修复DNA的紫外线诱导和化学诱导的损伤中起着共同的作用。

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