Thymidylate synthetase gene as a quantitative mutation marker in Chinese hamster cells

A quantitative mutation marker for cultured mammalian cells is presented which uses a selective medium containing folinic acid, aminopterin and thymidine (the 'FAT' medium) to select for mutants deficient in thymidylate synthetase (TS) activity. Optimization of FAT medium was carried out using Chinese hamster V79 cell lines having 3 levels of TS activity. By manipulating the concentration of folinic acid in FAT medium, TS-deficient mutants can be readily selected. TS mutation is inducible in a dose-dependent manner by either ethyl methanesulfonate or ultraviolet light irradiation. Expression time for TS mutation was also determined using two concentrations of ethyl methanesulfonate and found to be very short, being 1 or 2 days. This newly characterized TS mutation marker should be useful in the study of both spontaneous and induced mutagenesis.