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An improved system to examine forward mutations that occurred in the supF gene of Escherichia coli carried on a multicopy plasmid is described. The system was validated by measuring spontaneous mutations of supF plasmids propagated in wild-type, recA- and mutM- mutY- E. coli strains, the mutation frequencies of which were 1.3 x 10(-7), 6.3 x 10(-7) and 1.5 x 10(-6), respectively. Sequence analysis of the supF mutant plasmids revealed that G:C--&gt;T:A and G:C--&gt;C:G transversions dominated. This improved system allows rapid scoring and sequencing forward mutations in the supF gene, thus permitting its use as a genetic target for repair and mutagenesis studies in bacteria and mammalian cells.

An Improved System for Selection of Forward Mutations in an Escherichia coli supF Gene Carried by Plasmids.