Thompson L H, Busch D B, Brookman K, Mooney C L, Glaser D A
Proc Natl Acad Sci U S A. 1981 Jun;78(6):3734-7. doi: 10.1073/pnas.78.6.3734.
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.
通过遗传互补分析对中国仓鼠卵巢细胞的突变株进行了分析,这些突变株对紫外线杀伤和诱变表现出超敏反应,以确定不同基因位点的缺陷是否可能是共同细胞表型的基础。为便于快速筛选突变克隆,设计了一种程序,可根据聚乙二醇融合后抗紫外线细胞的频率评估推定互补情况。在测试的44个克隆中鉴定出四类。通过使用抗药标记物在紫外线突变体与野生型之间的杂交中选择杂交细胞,四类中的每一类的一个突变体表现为表型隐性。还从四个突变体的每对组合之间的杂交中分离出杂种。所有这些杂种对紫外线杀伤相对抗性,证实了互补类别。当用多环芳烃7-溴甲基苯并(a)蒽测试代表四个紫外线互补类别的突变体时,所有对组合再次出现互补。这些结果表明,每类突变体代表一种生化缺陷,在DNA中紫外线诱导和化学诱导损伤的修复中起共同作用。