Disruption and phenotypic analysis of six open reading frames from chromosome VII of Saccharomyces cerevisiae reveals one essential gene.
Six open reading frames (ORFs) located on chromosome VII of Saccharomyces cerevisiae (YGR205w, YGR210c, YGR211w, YGR241c, YGR243w and YGR244c) were disrupted in two different genetic backgrounds using short-flanking homology (SFH) gene replacement. Sporulation and tetrad analysis showed that YGR211w, recently identified as the yeast ZPR1 gene, is an essential gene. The other five genes are non-essential, and no phenotypes could be associated to their inactivation. Two of these genes have recently been further characterized: YGR241c (YAP1802) encodes a yeast adaptor protein and YGR244c (LSC2) encodes the beta-subunit of the succinyl-CoA ligase. For each ORF, a replacement cassette with long flanking regions homologous to the target locus was cloned in pUG7, and the cognate wild-type gene was cloned in pRS416.
Guerreiro
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Escola Superior de Tecnologia da Saúde de Lisboa, R. José Carlos dos Santos 7, P-2725-256 Lisboa, Portugal.
Rodrigues-Pousada
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D015825:Chromosomes, Fungal / Q000235:genetics*; D017353:Gene Deletion; D020043:Genes, Essential / Q000235:genetics*; D005800:Genes, Fungal / Q000235:genetics*; D016254:Mutagenesis, Insertional; D016366:Open Reading Frames / Q000235:genetics*; D010641:Phenotype; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development; D014170:Transformation, Genetic