PMID: 2494446Mar 1, 1989Paper

Molecular analysis of spontaneous mutations at the gpt locus in Chinese hamster ovary (AS52) cells

Mutation Research
K R Tindall, L F Stankowski

Abstract

AS52 cells are Chinese hamster ovary (CHO) cells that carry a single functional copy of the bacterial gpt gene and allow the isolation of 6-thioguanine-resistant (6TGr)mutants arising from mutation at the chromosally integrated gpt locus. The gpt locus in AS52 cells is extremely stable, giving rise to 6TGr mutants at frequencies comparable to the endogenous CHO hprt locus. In this study, we describe the spectrum of spontaneous mutations observed in AS52 cells by Southern blot and DNA sequence analyses. Using the polymerase chain reaction (PCR) and the Thermus aquaticus (Taq) polymerase, we have enzymatically amplified 6TGr mutant gpt sequences in vitro. The PCR product was then sequenced without further cloning manipulations to directly identify gpt structural gene mutations. Deletions predominant among the 62 spontaneous 6TGr-AS52 mutant clones analyzed in this study. Of these, 79% (49/62) of the mutations were identified as deletions either by Southern blotting, PCR amplification or DNA sequence analysis. Among these deletions is a predominant 3-base deletion that was observed in 31% (19/62) of the mutants. These data provide a basis for future comparisons of induced point mutational spectra derived in the AS52 cell line, and...Continue Reading

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Citations

Jan 16, 1999·Journal of Biochemical and Molecular Toxicology·M E Ariza, M V Williams
May 20, 1999·Teratogenesis, Carcinogenesis, and Mutagenesis·M J PlewaD Anderson
Jan 1, 1990·Teratogenesis, Carcinogenesis, and Mutagenesis·A AidooR H Heflich
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Aug 1, 1989·Mutation Research·J G Burkhart, H V Malling
Jun 6, 1994·The Science of the Total Environment·F E RossettoE Nieboer
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Apr 23, 2003·Environment International·Estelle FachMarshall V Williams
Sep 28, 1998·Biochimica Et Biophysica Acta·B C Baguley, L R Ferguson
Oct 20, 2001·Advances in Space Research : the Official Journal of the Committee on Space Research (COSPAR)·S R GondaB W Glickman
Apr 7, 2011·Environmental Science & Technology·Michael J PlewaWilliam A Mitch
Apr 12, 2006·Biological Chemistry·Akira Murakami, Hajime Ohigashi
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Oct 1, 1993·Environmental Health Perspectives·A W HsieP Hrelia
Jan 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·A B KulkarniS Karlsson
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Feb 24, 2006·Toxicology·Daniel Ballmaier, Bernd Epe
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