Figure 2

Complementation of hyporecombination phenotype of rec12-117 mutants by rec12⁺ cDNA and overexpression of rec12⁺ gene. (A) Recombination substrates. Intragenic recombination between the ade6-M26 and ade6-52 alleles was measured. (B) Complementation by rec12⁺ and full-length rec12⁺ cDNA. Assays were with rec12-117 strains harboring pREP42 inducible expression vector [32] constructs. Data are mean ± standard deviation from three separate experiments involving crosses of strains WSP1065 × WSP1067; WSP1066 × WSP1068; and WSP1823 × WSP1824. (C) Northern blot analysis of rec12⁺ gene expression induced from strains harboring low-, middle-, and high-expression versions of pREP vectors. Data were obtained using strains WSP1058; WSP1066; and WSP1074. (D) Effect of rec12⁺ expression level upon recombination in rec12-117 strain. Recombinant frequency data are the mean ± standard deviation from three separate experiments; expression levels are β-galactosidase levels of similar constructs under non-inducing and inducing conditions [32]. Relative rec12⁺ expression levels were: pREP2^u (^u = uninduced; ⁱ = induced) < pREP82-rec12^+,u < pREP42-rec12^+,u < pREP82-rec12^+,i < pREP42-rec12^+,i < pREP2-rec12^+,u < pREP2-rec12^+,i. Data were obtained from crosses of strains WSP1073 × WSP1075; WSP1058 × WSP1060; WSP1066 × WSP1068; and WSP1074 × WSP1076.