FIGURE 2.
The E. coli Lac assay for stress-induced mutagenesis. Above: schematic representation of the strain used in the Lac assay. This strain bears a ~200 kb F′ conjugative plasmid carrying the mutant lacIZ33 allele, a lacI-lacZ fusion gene with a +1 frameshift mutation. Below: generation-dependent mutation events occurring during growth of the culture prior to plating on lactose medium are detected as Lac+ colonies present on about day 2. During subsequent days of incubation, stress-induced Lac+ colonies accumulate, and include both Lac+ point mutants with a compensatory frameshift mutation or lac-amplified cells, with 20 to 50 copies of DNA spanning the lac gene, which provides sufficient beta-galactosidase activity for growth without a frameshift reversion mutation (Hastings et al., 2004).