Derivation of Tk- Clones from Revertant Tk+ Mammalian Cells

AUTOR(ES)

Roufa, D. J.

RESUMO

In order to obtain a large collection of Chinese hamster cell clones defective in thymidine kinase (TK-), BrdUr selection experiments have been performed on wild-type and revertant TK+ cell lines. No clones (< 10-9) were obtained from the wild-type TK+ cell line by single-step selection. In contrast, revertant TK+ clones readily gave rise to stable TK- derivatives (1 - 2 x 10-4). Both wild-type and revertant TK+ clones spontaneously yielded 8-AGr colonies with the same frequency (1 - 5 x 10-6), suggesting that the differences between wild-type and revertant cell lines specifically affected selection of the TK- phenotype. The increased frequency of TK- clones reflects perhaps the number (ploidy) or character of the autosomal TK loci in TK+ revertants, or perhaps the mechanisms which regulate expression of the TK genes. Several mutagens, EMS, MNNG and UV, stimulated the TK+ revertants' frequency of TK- subclones only slightly (< 3-fold). Biochemical and genetic data indicated that the TK- clones derived from one revertant are phenotypically different. The phenotypes displayed by these cell lines are stable and do not depend upon the continued presence of the selective agent.

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