title: Unique pattern of point mutations arising after gene transfer into mammalian cells.
abstract: We have used a simian virus 40 (SV40)-based shuttle vector, pZ189, to analyze the sequence specificity of spontaneous point mutations that arise after transfection of this vector into monkey cells. The majority of the mutants which we studied had multiple base substitutions (mostly G-C----A-T transitions and G-C----T-A transversions) within the 160-bp region sequenced. Almost all of the mutations occurred in the right-hand G-C bp of one of the two following sequences, 5'-TC-3':3'-AG-5' or 5'-CC-3':3'-GG-5'. We postulate that these mutations result from DNA replication infidelity occurring during repair of the transfected DNA which has been damaged by cellular nucleases. The sequence specificity of the mutations suggests an effect of the following nucleotide on misincorporation wherein A (or less frequently T) is preferentially misincorporated opposite C when the next nucleotide inserted is A (or less frequently G). Our results support the utility of the shuttle vector as a model in studies on gene transfer and document the extreme plasticity of DNA transfected into mammalian cells.
mesh_terms: D000818:Animals; D001482:Base Composition; D001483:Base Sequence; D002460:Cell Line; D004261:DNA Replication; D005822:Genetic Vectors; D009154:Mutation; D010957:Plasmids; D013539:Simian virus 40 / Q000235:genetics; D014162:Transfection