Generation of null alleles for the functional analysis of six genes from the right arm of Saccharomyces cerevisiae chromosome II.

ABSTRACT

Using PCR-ligated long flanking homology cassettes, null alleles of six open reading frames (ORFs) from chromosome II have been created in Saccharomyces cerevisiae. Deletants were constructed in three genetic backgrounds FY1679, W303 and CEN.PK2. Tetrad analysis of heterozygous deletants revealed that none of the ORFs is essential for vegetative growth. Basic phenotypic analysis of haploid deletants showed that deletion of the YBR283c ORF causes a slight growth defect at 30 degrees C and 37 degrees C on glycerol-complete, glucose-complete, and glucose-minimal media only in the FY1679 and W303 backgrounds. Transformation of these deletants with the corresponding cognate gene in a centromeric plasmid complements the defects. Deletion of the YBR287w ORF leads to poor growth on glucose-minimal medium at 15 degrees C in the FY 1679 background. None of the six ORFs seems to be involved in mating or sporulation.