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Mutagenicity of an antitumor protein, neocarzinostatin, in Escherichia coli. |
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| Authors: | K Tatsumi H Nishioka |
| Institution: | Environmental and Medical Sciences Division, AERE Harwell, Oxfordshire OX11 ORA, England |
| Abstract: | 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. |
| Keywords: | HX hypoxanthine HGPRT HX-guanine phosphoribosyl transferase F12FCM5 medium F12 containing 5% dialyzed fetal calf serum F12FCM10 medium F12 containing 10% dialyzed fetal calf serum TG 6-thioguanine (2-amino 6-mercaptourine) TG medium HX-free medium F12 containing 5% dialyzed fetal calf serum and 10 μM TG AP aminopterin AP medium medium F12FCM10 containing 1 μM AP SDS sodium dodecyl sulfate TCA trichloroacetic acid EMS ethyl methanesulfonate |
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