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 genetic backgrounds. Six open reading frames (ORFs) from chromosome IV, YDR013w, YDR014w, YDR015c, YDR018c, YDR020c and YDR021w, were disrupted using the KanMX4 marker and PCR-targeting with long flanking regions homologous (LFH) to the target locus. The deletants were verified at the molecular level, using PCR and Southern analysis. Sporulation and tetrad analysis revealed that ORFs YDR013w and YDR021w (also known as FAL1) are essential genes. Microscopical observations showed that ydr013wDelta haploid cells were blocked after one or two cell cycles and presented heterogeneous bud sizes. The ydr021wDelta haploid cells gave rise to microcolonies of about 20 cells. The other four ORFs are non-essential. Basic phenotypic analysis of the non-lethal deletant strains did not reveal any significant differences in cell morphology, growth on different media and temperatures, sporulation and mating efficiency between parental and mutant strains in the FY1679 background.
Dardalhon
M
M
Institut Curie, Section de Recherche, CNRS UMR 2027, Centre Universitaire, Bat. 110, 91405 Orsay Cedex, France. Michele.Dardalhon@curie.u-psud.fr
Magaña-Schwencke
N
N
Averbeck
D
D
D015139:Blotting, Southern; D015825:Chromosomes, Fungal / Q000235:genetics; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology; D017353:Gene Deletion; D005800:Genes, Fungal; D016366:Open Reading Frames / Q000235:genetics*; D010641:Phenotype; D016133:Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*
D005656:Fungal Proteins