Quantitative plating assay for suppression of rec104-8 by specific alleles of REC102 a

StrainRelevant genotypePlasmidbSporulation %No. of surviving asci/totalcRFd (103) Avg fold increasee
LS5-1 rec104-8 pRS426 (vector)272/300.220.0400.009
LS5-1 rec104-8 pJK22 (hcREC102)2811/302.3 (10)0.21 (5.3)0.17 (18)11.5
LS5-1 rec104-8 pLS27 (REC102-35)4113/302.9 (13)0.30 (7.5)0.20 (22)14.2
LS5-1 rec104-8 pLS30 (REC102-48)7822/308.9 (40)4.1 (102)1.2 (133)92.1
  • a Quantitative plating assay for recombination. The LS5-1 diploid was transformed with the indicated plasmid, and transformants were sporulated at the nonpermissive temperature (35°C). Sporulated cells were plated on media diagnostic for recombination, as well as complete media, to determine the frequency of recombination. All experiments were done on the same day. The results shown are averages of three independent cultures.

  • b The relevant gene contained in the plasmid is in parentheses. The plasmids containing REC102-35 and REC102-48 were CEN plasmids.

  • c The viability of meiotic products was monitored by examining 30 asci to determine how many could make a viable colony. At least one spore must be alive for an ascus to make a colony.

  • d Recombination frequency (RF) is calculated as the number of recombinants divided by the total number of cells plated. The values in parentheses are fold increases relative to the vector control (pRS426). Canr colonies are an indirect measure of crossing over, since recombination is required for haploidization and viability.

  • e The average increase is the average, over all three loci, of the relative amount of recombination compared to the cells containing the vector.