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.