Disruption and basic phenotypic analysis of six novel genes from the left arm of chromosome XIV ofSaccharomyces cerevisiae

✍ Scribed by Dueñas, Encarnación; Revuelta, José Luis; Del Rey, Francisco; Vázquez de Aldana, Carlos R.

• Publisher: John Wiley and Sons
• Year: 1999
• Tongue: English
• Weight: 337 KB
• Volume: 15
• Category: Article
• ISSN: 0749-503X
• DOI: 10.1002/(sici)1097-0061(19990115)15:1<63::aid-yea338>3.0.co;2-7

✦ Synopsis

We describe here the construction of six deletion mutants and their basic phenotypic analysis in three different backgrounds. The six genes were disrupted in three diploid strains (FY1679, W303 and CEN.PK2) by the long flanking homology (LFH) method (Wach, 1996). Transformants were selected as geneticin (G418)-resistant colonies and correct integration of the kanMX4 cassette was checked by colony PCR. Following sporulation of the heterozygous diploids, tetrads were dissected and scored for segregation of G418-resistance and auxotrophic markers. One of the six ORFs (YNL158w) corresponds to an essential gene which has no homology with other genes present in the databases and has two predicted transmembrane domains. Growth tests performed on different media at 15 C, 30 C or 37 C with haploid deletants of the five non-essential genes revealed no apparent phenotype in any of them.