Use of sulfite resistance in Saccharomyces cerevisiae as a dominant selectable marker.
Two S. cerevisiae genes were found to exhibit dominant phenotypes useful for selecting transformants of industrial and laboratory strains of S. cerevisiae. FZF1-4, which confers sulfite resistance, was originally isolated and identified as RSU1-4, but the two genes are shown here to be allelic. Cysteine 57 in wild-type Fzf1p was found to be replaced by tyrosine in Fzf1-4p. Multicopy SSU1, which also confers sulfite resistance, was found to be somewhat less efficient. In both cases, a period of outgrowth in non-selective medium following transformation was found to be necessary. The number of transformants obtained was found to be strain-dependent, and also to depend on the sulfite concentration used during selection. Undesirable background growth of non-transformants was not observed at cell densities as high as 2.5 x 10(7)/plate. In two ura3 laboratory strains where selection for URA3 was applied independently of that for sulfite, the transformation efficiency for sulfite resistance was about 50% that for uracil prototrophy.
Park
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Department of Food Science and Technology, Oregon State University, Corvallis 97331-6602, USA.
Lopez
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Bakalinsky
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D027321:Anion Transport Proteins; D004352:Drug Resistance, Microbial / Q000235:genetics*; D005656:Fungal Proteins / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D005799:Genes, Dominant; D005816:Genetic Complementation Test; D005819:Genetic Markers; D008565:Membrane Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins; D012641:Selection, Genetic; D017422:Sequence Analysis, DNA; D013447:Sulfites / Q000494:pharmacology*; D014157:Transcription Factors; D014170:Transformation, Genetic; D016335:Zinc Fingers / Q000235:genetics
D027321:Anion Transport Proteins; C090870:FZF1 protein, S cerevisiae; D005656:Fungal Proteins; D005819:Genetic Markers; D008565:Membrane Proteins; D029701:Saccharomyces cerevisiae Proteins; C108311:Ssu1 protein, S cerevisiae; D013447:Sulfites; D014157:Transcription Factors