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, YDL073w, YDL076c, YDL077c and YDL080c, were disrupted using the long flanking homology technique (LFH) to replace each target locus with the KanMX4 selection marker. We have also constructed plasmids containing replacement cassettes (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 requirements. However, disruption of YDL077c (also known as VAM6) exhibited an slow growth phenotype in minimal media and also in rich media containing glycerol as a carbon source. The homozygous disruptant diploid corresponding to this gene also failed to sporulate.
Blasco
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Instituto de Biomedicina de Valencia (CSIC), Jaime Roig 11, 46010-Valencia, Spain.
Sanz
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D015825:Chromosomes, Fungal / Q000737:chemistry / Q000235:genetics*; D003001:Cloning, Molecular; D017931:DNA Primers / Q000737:chemistry; D004271:DNA, Fungal / Q000737:chemistry; D016366:Open Reading Frames / Q000235:genetics*; D010641:Phenotype; D010957:Plasmids; D016133:Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000235:genetics*
D017931:DNA Primers; D004271:DNA, Fungal