Abstract
Four independent mutants were isolated from mutagenized cultures of CHO cells by sib selection on the basis of resistance to a low concentration (2.6 × 10−5M) of BrdU. All four lines were stable, but all had about 100% of the wild-type (WT) specific activity of thymidine kinase (TK). None of the four yielded derivatives resistant to a high level of BrdU (2 × 10−4M) in one step even after mutagenesis, but variants resistant to 4–6 × 10−5M BrdU could be isolated at frequencies of about 2 × 10−5/cell. At frequencies of 10−4−10−5, the second-step mutants gave colonies resistant to 2 × 10−4 M BrdU. The second and third steps of resistance were correlated with partial and complete reduction, respectively, in the specific activity of TK, suggesting that the variants may be genotypically heterozygous and homozygous-negative at the tk locus. The first step of BrdU resistance was dominant and appeared to result from a mutation in the gene for ribonucleotide reductase, since in vitro assays on partially purified preparations showed that the reductase activity in mutant cells was less sensitive to BrdUTP than that in WT cells.
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Bradley, W.E.C., Dinelle, C., Charron, J. et al. Bromodeoxyuridine resistance in CHO cells occurs in three discrete steps. Somat Cell Mol Genet 8, 207–222 (1982). https://doi.org/10.1007/BF01538678
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DOI: https://doi.org/10.1007/BF01538678