Disruption and basic functional analysis of five chromosome X novel ORFs ofSaccharomyces cerevisiae revealsYJL125c as an essential gene for vegetative growth

✍ Scribed by Jaquet, Laurence; Jauniaux, Jean-Claude

• Publisher: John Wiley and Sons
• Year: 1999
• Tongue: English
• Weight: 388 KB
• Volume: 15
• Category: Article
• ISSN: 0749-503X
• DOI: 10.1002/(sici)1097-0061(19990115)15:1<51::aid-yea330>3.0.co;2-1

✦ Synopsis

We describe the disruption and basic functional analysis of five novel open reading frames (ORFs) discovered during the sequencing of the Saccharomyces cerevisiae genome: YJL118w, YJL122w, YJL123c, YJL124c, YJL125c, located on chromosome X. Disruptions have been realized using the long-flanking homology-PCR replacement strategy (LFH-PCR;Wach et al., 1996) in the FY1679 diploid strain. Sporulation and tetrad analysis of these heterozygous deletants were performed, as well as a functional analysis on the haploid deleted strains: different growth conditions (complete glucose and glycerol, minimal media) at three temperatures 15, 30 and 37 C were tested. Analysis revealed YJL125c as an essential gene; the four other ORFs were non-essential and showed no particular phenotype. In addition, the five kanMX4 disruption cassettes were cloned in pUG7 vector. Finally, the five ORFs with their promoter and terminator regions were cloned in the centromeric yeast vector pRS416. The vectors containing the disruption cassettes, the cognate wild-type genes, as well as the deletant yeast strains are available at the EU EUROFAN (EUROSCARF, Frankfurt, DE) genetic and stock centre.