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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)

Quantification and analysis of reverse mutations at the hgprt locus in Chinese hamster ovary cells.

We describe an assay for the quantification of reverse mutations at the hypoxanthine-guanine phosphoribosyltransferase (hgprt) locus in Chinese hamster ovary cells utilizing the selective agent L-azaserine (AS). Conditions are defined in terms of optimal AS concentration, cell density, and phenotypic expression time. After treatment, replicate cultures of 10(6) cells are allowed a 48-h phenotypic expression time in 100-mm plates. AS (10 muM) is then added directly to the growing culture and AS-resistant (ASr) cells form visible colonies. This assay is used to quantify ICR-191-, ICR-170-, and N-ethyl-N-nitrosourea-induced reversion of independently isolated HGPRT- clones. The ASr phenotype is characterized both physiologically and biochemically. All ASr clones isolated are stably resistant to AS and aminopterin but sensitive to 6-thioguanine. They also have re-expressed HGPRT enzyme. In addition, several revertants are shown to contain altered HGPRT. The data provide further evidence that ICR-191 and ICR-170 cause structural gene mutations in mammalian cells and also suggest that ICR-191, ICR-170, and N-ethyl-N-nitrosourea induce similar types of mutations in Chinese hamster ovary cells.[1]


  1. Quantification and analysis of reverse mutations at the hgprt locus in Chinese hamster ovary cells. Fuscoe, J.C., O'Neill, J.P., Machanoff, R., Hsie, A.W. Mutat. Res. (1982) [Pubmed]
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