Deletion of six open reading frames from the left arm of chromosome IV of Saccharomyces cerevisiae

The construction of six deletion mutants of Saccharomyces cerevisiae and th eir basic phenotypic characterization are described. Open reading frames YD L148c, YDL109c, YDL021w, YDL019c, YDL018c and YDL015c from the left arm of chromosome IV were deleted using a polymerase chain reaction (PCR)-based di sruption technique, introducing the kanMX4 resistance marker into the respe ctive genes. Gene replacement cassettes (pYORCs) for use in other strain ba ckgrounds were cloned by PCR using DNA templates from haploid or diploid de letion mutants, and inserted into episomal plasmids. Cognate clones of all six ORFs were obtained by gap repair. Deletions were carried out in diploid cells and, after sporulation, yielded four viable spores for clones disrup ted in YDL109c, YDL021w, YDL019c and YDL018c. Spores harbouring disruptions in ORFs YDL148c and YDL015c germinated but underwent only a few divisions before ceasing growth, suggesting that the respective genes are essential f or vegetative growth on YPD complete media. The other deletion mutants grew like wild-type at different temperatures and on different carbon sources. A brief computational analysis of the six ORFs studied in this work is pres ented. Copyright (C) 1999 John Wiley & Sons, Ltd.