Complementation of the DNA-repair defect in a CHO mutant by human DNA that lacks highly abundant repetitive sequences

Mutation Research/DNA Repair Reports Pub Date : 1988-11-01 DOI:10.1016/0167-8817(88)90022-3

Ann M. Dulhanty , Jaime S. Rubin , Gordon F. Whitmore

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引用次数: 12

Abstract

Recently, two human DNA-repair genes have been cloned which complement the defects in complementation groups 1 and 2 of the CHO mutants which are sensitive to ultraviolet light and deficient in the incision step of excision repair. Here we report human gene transfer-mediated complementation of a group 4 CHO mutant sensitive to ultraviolet light and mitomycin C (MMC). The transfectants generated by transfecting human DNA into the repair-deficient cell line demonstrate the repair-proficient phenotype, as they have wild-type levels of resistance to UV light and MMC and are competent in performing the incision step of excision erpair in response to UV irradiation. 3 of the 8 transfectants isolated display no detectable human repetitive sequences, while the other 5 contain varying amounts of human repetitive DNA. As the evidence suggests that all of the transfectants are repair-proficient as a result of the uptake of humand DNA, we conclude that the human gene that complements the repair defect in group 4 CHO mutants contains no highly abundant human repetitive sequences. This imposes the necessity of developing cloning strategies involving the identification of sequences that flank the gene.

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缺乏高度丰富重复序列的人DNA对CHO突变体中DNA修复缺陷的补充

最近，克隆了两个人类dna修复基因，它们弥补了CHO突变体中对紫外光敏感且在切除修复的切口步骤中缺乏的互补组1和2的缺陷。在这里，我们报道了人类基因转移介导的对紫外线和丝裂霉素C (MMC)敏感的4组CHO突变体的互补。通过将人类DNA转染到修复缺陷细胞系中产生的转染物显示出修复能力强的表型，因为它们对紫外线和MMC具有野生型的抗性，并且能够在紫外线照射下执行切除修复的切口步骤。8个分离的转染物中有3个没有显示可检测的人类重复序列，而其他5个含有不同数量的人类重复DNA。有证据表明，由于摄取人类DNA，所有的转染物都具有修复能力，因此我们得出结论，补充第4组CHO突变体修复缺陷的人类基因不包含高度丰富的人类重复序列。这就要求有必要发展克隆策略，包括鉴定基因侧面的序列。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Mutation Research/DNA Repair Reports

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