Extragenic suppression of tra1-L3733A. A. CY4018 (tra1-L3733A, LA) was plated onto YPD media containing 4% ethanol (EtOH) at 30°C. Growing colonies were in succession: colony purified on media containing ethanol, plated on YPD at 30°C, and retested on media containing ethanol. The 11 strains passing this selection (CY5579, es2; CY5580, es12; CY5581, es35; etc.), CY2706 (TRA1), and CY4018 (tra1-L3733A) were grown to stationary phase, serially diluted and plated onto YPD or YPD containing 4% ethanol at 30°C. B. The es2, es38 and es41 alleles are recessive. Strains CY5579 (es2 tra1-L3733A), CY5584 (es38 tra1-L3733A) and CY5587 (es41 tra1-L3733A) were mated with CY5522 (tra1-L3733A) to generate diploid strains homozygous for tra1-L3733A and heterozygous for each suppressor. Serial dilutions of these strains as well as the diploid wild-type strain CY5557 and the homozygous tra1-L3733A strain CY5558 were spotted onto the indicated plates. C. es2 forms an independent complementation group. Serial dilutions of diploid strains arising from crosses of haploid strains CY5579 with CY5522 (es2 x ES2), CY5579 with CY5666 (es2 x es2), CY5666 with CY5584 (es2 x es38), and CY5666 with CY5587 (es2 x es41) were spotted onto the indicated plates.
Kvas et al. BMC Genetics 2012 13:19 doi:10.1186/1471-2156-13-19