“…The following strains of a similar design, rps5-K41A Mat a
his3-1, leu2-0, ura3-0 , rps5::kanMX , < yrps5-K41A ; LEU2, 2μ >, rps5-F43G Mat a
his3-1, leu2-0, ura3-0 , rps5::kanMX , < yrps5-F43G ; LEU2, 2μ >, rps5-K45A Mat a
his3-1, leu2-0, ura3-0 , rps5::kanMX , < yrps5-K45A ; LEU2, 2μ >, expressing yeast rps5 variants with the indicated amino acid substitutions, were obtained as follows: point mutations were first introduced into the RPS5 gene in pTEF_yS5 plasmid (10) using side-directed mutagenesis and the following primers 5′-CAAACCGAGATTGCGTTGTTCAAC-3′ forward and 5′-GTTGAACAACGCAATCTCGGTTTG-3′ reverse (pTEF_yS5-K41A); 5′-GAGATTAAGTTGGGCAACAAATGGTC-3′ forward and 5′-GACCATTTGT TGCCCAACTTAATCTC-3′ reverse (pTEF_yS5-F43G); and 5′-AGTTGTTCAACGCATGGTCTTTTG-3′ forward and 5′-CAAAAGACCATGCGTTGAACAACT-3′ reverse (pTEF_yS5-K45A). The resultant pTEF_yS5 plasmids carrying mutant RPS5 were transformed into the heterozygous diploid BY4743 [4741/4742] MAT a /MATα, his3-1/his3-1, leu2-0/leu2-0, lys2-0/+, met15-0/+, ura3-0/ura3-0, RPS5/rps5::kanMX .…”