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A Quantitative Assay Of Mutation Induction At The Hypoxanthine-guanine Phosphoribosyl Transferase Locus In Chinese Hamster Ovary Cells (CHO/HGPRT System): Development And Definition Of The System.
J. O'Neill, P. Brimer, R. Machanoff, G. P. Hirsch, A. Hsie
Published 1977 · Biology, Medicine
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An assay is described for the measurement of mutation induction at the hypoxanthine-guanine phosphoribosyl transferase (HGPRT) locus in Chinese hamster ovary (CHO) cells utilizing resistance to 6-thioguanine (TG). Optimal selection conditions are defined for such parameters as phenotypic expression time prior to selection, and TG concentration and cell density which permits maximum mutant recovery. The nature of the TG-resistant mutants is characterized by several physiological and biochemical methods. The data demonstrate that more than 98% of the mutant clones isolated by this selection procedure contain altered HGPRTase activity. The CHO/HGPRT system thus shows the specificity necessary for a specific gene locus mutational assay.
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