Transgenic Chinese hamster V79 cell lines which exhibit variable levels of gpt mutagenesis.
The Escherichia coli gpt gene coding for xanthine-guanine phosphoribosyl transferase has been stably transfected into HPRT- Chinese hamster V79 cells. Several gpt- cell lines have been established, which retain the sequence(s) even after long-term culture without selection for gpt. Each cell line exhibits a characteristic spontaneous mutation frequency (10(-5) to 10(-2)) in 6-thioguanine (6TG) selection. While spontaneous mutagenesis to gpt- occurs rather frequently for most cell lines, it cannot be correlated with either the number of plasmid integration sites or deletion of the plasmid sequence(s). One transgenic cell line (g12), which continuously maintains a low spontaneous mutation frequency (approximately 3 x 10(-5)), was used in comparative mutagenesis studies with wild-type V79 cells (gpt vs. hprt). Alkylating agents such as N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and beta-propiolactone (BPL) are shown to be equally toxic and mutagenic in both g12 and V79 cells. UV and X-rays are also equally toxic to both cell lines. The gpt locus of the g12 transfectants, however, is two to three times more sensitive to UV and 2.5-4.5 times more sensitive to X-ray mutagenesis than the endogenous hprt of wild-type V79 cells. The data presented here suggests that g12 cells may be useful to study mammalian mutagenesis by agents which yield limited response at the hprt locus. Future studies with these transgenic cells and other transgenic lines are planned to compare the mutability and repair of the same gene (gpt) at different integration sites in mammalian cells.
Klein
C B
CB
Institute of Environmental Medicine, New York University Medical Center, New York, New York.
Rossman
T G
TG
D000818:Animals; D015139:Blotting, Southern; D002460:Cell Line; D006224:Cricetinae; D004926:Escherichia coli; D005798:Genes, Bacterial; D007041:Hypoxanthine Phosphoribosyltransferase / Q000378:metabolism; D008769:Methylnitronitrosoguanidine / Q000633:toxicity; D009152:Mutagenicity Tests / Q000379:methods*; D009154:Mutation; D010430:Pentosyltransferases / Q000235:genetics*; D011420:Propiolactone / Q000494:pharmacology; D014162:Transfection; D014466:Ultraviolet Rays; D014965:X-Rays
D008769:Methylnitronitrosoguanidine; D011420:Propiolactone; D010430:Pentosyltransferases; C024827:xanthine phosphoribosyltransferase; D007041:Hypoxanthine Phosphoribosyltransferase