Disruption and basic phenotypic analysis of six novel genes from the rightarm of chromosome XII of Saccharomyces cerevisiae

Sis open reading frames (ORFs) of unknown function from the right arm of Sa ccharomyces cerevisiae chromosome XII were deleted in two genetic backgroun ds by disruption cassettes with regions of short flanking homology. This wo rk was carried out,within the framework of the EUROFAN consortium. The SFH disruption cassettes, obtained by PCR, were made by amplification of the ka nMX marker module with oligonucleotides containing approximately 40 bp of h omology to either the promoter or translation terminator regions of the rel evant ORF. Transformants resistant to geneticin (G418) were selected. The S FH disruption cassettes were cloned into a bacterial vector. Each cognate g ene was also cloned into a yeast centromeric plasmid. Sporulation and tetra d analysis of the disrupted heterozygous strains revealed that ORF YLR153c (now known as ACS2) is essential. Basic phenotypic analysis mas performed o n haploid deletants of both mating types of the five non-essential ORFs, YL R082e (now known as SRL2), YLR149c, YLR151c, YLR152c and YLR154c. Plate gro wth tests on different media at 15 degreesC, 30 degreesC and 37 degreesC di d not reveal any significant differences between parental and mutant cells. Mating and sporulation efficiencies were not affected in any of the viable disruptants as compared to wild-type cells. Copyright (C) 2001 John Wiley & Sons, Ltd.