title: Use of mutated PDR3 gene as a dominant selectable marker in transformation of prototrophic yeast strains.
abstract: For successful transformation of prototrophic industrial yeast strains dominant selectable markers are necessary. In the present study we show the applicability of a selection system based on the phenotype of multidrug resistance. The mutant pdr3-9 allele on centromeric or episomal vector, encoding a more efficient transcriptional activator with Y276H amino acid substitution, was used as a dominant selectable marker for selection of transformants. The pdr3-9 allele conferred resistance of transformed cells to cycloheximide, chloramphenicol, mucidin and oligomycin both in the absence and in the presence of a chromosomal copy of the PDR3 gene. Both multicopy YEp352/pdr3-9 and centromeric pFL38/pdr3-9 vectors bearing the mutant pdr3-9 allele have proved to be a valuable tool for a direct selection of transformants of industrial strains of Saccharomyces cerevisiae.
authors.affiliation: Department of Microbiology and Virology, Faculty of Science, Comenius University, Bratislava, Slovakia.
authors.forename: D
authors.initials: D
authors.lastname: Lacková
authors.forename: J
authors.initials: J
authors.lastname: Subík
mesh_terms: D000935:Antifungal Agents / Q000494:pharmacology; D004268:DNA-Binding Proteins / Q000235:genetics*; D004352:Drug Resistance, Microbial / Q000235:genetics; D018432:Drug Resistance, Multiple / Q000235:genetics; D005799:Genes, Dominant; D005800:Genes, Fungal; D005819:Genetic Markers; D008826:Microbial Sensitivity Tests; D009154:Mutation; D010957:Plasmids / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors / Q000235:genetics*; D014170:Transformation, Genetic; D014498:Uracil / Q000378:metabolism
chemical_list: D000935:Antifungal Agents; D004268:DNA-Binding Proteins; D005819:Genetic Markers; C089261:PDR3 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D014498:Uracil