Disruption and functional analysis of six ORFs on chromosome IV: YDL053c, YDL072c, YDL073w, YDL076c, YDL077c and YDL080c

Six open reading frames (ORFs) from chromosome IV, YDL053c, YDL072c, YDL073 w, YDL076c, YDL077c and YDL080c, mere disrupted using the long flanking hom ology technique (LFH) to replace each target locus with the KanMX4 selectio n marker. We have also constructed plasmids containing replacement cassette s (pYORC) and the cognate clones (pYCG) for each ORF. Disruption of five of the ORFs-YDL053c, YDL072c, YDL073w, YDL076c and YDL080c (THI3)-resulted in no distinctive phenotype with respect to temperature or nutritional requir ements. However, disruption of YDL077c (also known as VAM6) exhibited an sl ow growth phenotype in minimal media and also in rich media containing glyc erol as a carbon source. The homozygous disruptant diploid corresponding to this gene also failed to sporulate. Copyright (C) 2000 John Wiley & Sons, Ltd.