High mutation frequency in DNA transfected into mammalian cells.
The lacI gene of Escherichia coli was used to score mutation in mammalian cells of simian virus 40-based recombinant DNA vectors that provide for replication and selection in both bacterial and mammalian cells. Plasmid DNA was introduced into COS7 simian cells by DEAE-dextran transfection, allowed to replicate in the mammalian cells, and then returned to E. coli for analysis. Mutants in lacI were observed at frequencies of one to several percent, compared with a spontaneous mutation rate in E. coli of less than 10(-5). The lesions include a large number of base substitutions, in addition to deletions, duplications, and more complex rearrangements, including insertion into the plasmid of sequences originating in the host genome. We discuss possible sources of the high mutation frequency and its implications for experiments involving DNA transfer.
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D004260:DNA Repair; D004261:DNA Replication; D004274:DNA, Recombinant; D007763:Lac Operon; D009154:Mutation; D010957:Plasmids; D014162:Transfection
D004274:DNA, Recombinant