Disruption and functional analysis of six ORFs on chromosome IV: YDR013w, YDR014w, YDR015c, YDR018c, YDR020c, YDR021w (FAL1)

The disruption of six novel yeast genes has been realized in two generic ba ckgrounds. Six open reading frames (ORFs) from chromosome IV, YDR013w, YDR0 14w, YDR015c, YDR018c, YDR020c and YDR021w, were disrupted using the KanMX4 marker and PCR-targeting with long flanking regions homologous (LFH) to th e target locus. The deletants were verified at the molecular level, using P CR and Southern analysis. Sporulation and tetrad analysis revealed that ORF s YDR013w and YDR021w (also known as FAL1) are essential genes. Microscopic al observations showed that ydr013w Delta haploid cells were blocked after one or two cell cycles and presented heterogeneous bud sizes. The ydr021w D elta haploid cells gave rise to microcolonies of about 20 cells. The other four ORFs are non-essential. Basic phenotypic analysis of the non-lethal de letant strains did not reveal any significant differences in cell morpholog y, growth on different media and temperatures, sporulation and mating effic iency between parental and mutant strains in the FY1679 background. Copyrig ht (C) 2000 John Wiley & Sons, Ltd.