||Utilization of Trifluorothymidine (TFT) to Select for Thymidine Kinase-Deficient Mutants from L5178Y/TK Mouse Lymphoma Cells.
Moore-Brown, M. M. ;
Clive, D. ;
Howard, B. E. ;
Batson, A. G. ;
Johnson, K. O. ;
||Health Effects Research Lab., Research Triangle Park, NC. ;Burroughs Wellcome Co., Research Triangle Park, NC. ;North Carolina Univ. at Chapel Hill.
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Trifluorothymidine (TFT), a thymidine analog, was analyzed for its ability to select for thymidine kinase-deficient (TK-/-) mutants. In comparison with BUdR, the traditional selective agent for TK-/- cells, it was determined that TFT at 1/50th the dose (1 micrograms/ml vs. 50 micrograms/ml) is a more effective and versatile selective agent for TK-/- mutants arising from the TKt/- 3.7.2C heterozygote of L5178Y mouse lymphoma cells. Since TFT acts more rapidly than BUdR, it can be utilized in procedures (such as the analysis of the phenotypic lag) requiring the fast arrest of cell division. Reconstruction analyses of effective TK-/- mutant recovery indicate that TFT can be used to recover mutants from significantly higher densities of TIt/- cells than can BUdR. In addition, TK -/- mutants can attain larger colony size in TFT than in BUdR where severe stunting of growth occurs at high TK-/- cell densities. 190 of 194 isolated TFT-resistant large and small colony mutants (both spontaneous and induced).