DNA base sequence changes induced by ethyl methanesulfonate in a chromosomally integrated shuttle vector gene in mouse cells

Abstract

We have analyzed the specificity of mutations induced by ethyl methanesulfonate (EtMes) in mouse cells carrying a selectable bacterial gene. The target gene was the Escherichia coli gpt gene contained within a retroviral shuttle vector integrated into mouse chromosomal DNA. Following mutagenesis by EtMes, cells with mutations in the gpt gene were selected as resistant to 6-thioguanine. Shuttle vector sequences were recovered from the mutant cell lines following fusion with monkey COS cells and introduced into bacteria as part of a bacterial plasmid. The DNA base sequences of the mutant genes were directly determined from plasmid DNA. All of the EtMes-induced mutations involving single base changes were found to be G:C to A:T transitions.