Figure 7

Homologous recombination efficiency in wild type and mutant strains. (A) Schematic of the homologous recombination assay. A PCR product spanning the deleted region of the genomic his3Δ1 allele was transformed into competent yeast cells. Homologous recombination of the PCR fragment with the genomic allele generates His-positive colonies. The hatched box indicated the Hin dIII (H) fragment deleted in the his3Δ1 allele. Numbers refer to base pair locations within the HIS3 open reading frame. (B) Recombination rate, relative to wild type, was measured as the efficiency of His-positive prototroph formation (cfus/cells transformed) at 30 or 34°C. The numbers represent the average of three transformation reactions and the standard deviation is given for each value. Efficiency of transformation was normalized to the transformation efficiency of a HIS3 marked 2κ plasmid compared with wild type. (C) Graphical representation of numbers given in (B). Strains are as follows: wild type JKc1046; rfx1Δ JKc1532; sml1Δ JKc1533.