Abstract
Strains of Saccharomyces cerevisiae deleted in the NADPH-cytochrome P450 reductase gene by transplacement are 200-fold more sensitive to ketoconazole, an inhibitor of the cytochrome P450 lanosterol 14 alpha-demethylase. Resistance is restored through complementation by the plasmid-borne wild type gene from either S. cerevisiae or Candida tropicalis. Neither Southern hybridization nor Western immunoblot techniques provided evidence for a second NADPH-cytochrome P450 reductase gene, suggesting that an alternate pathway may provide for the functions of this reductase in S. cerevisiae.
Publication types
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Blotting, Western
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Candida / genetics
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Cytochrome P-450 Enzyme System*
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DNA Restriction Enzymes
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DNA, Fungal / isolation & purification
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Drug Resistance, Microbial / genetics
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Genes, Fungal*
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Ketoconazole / pharmacology*
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Leucine / physiology
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NADPH-Ferrihemoprotein Reductase / genetics*
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Nucleic Acid Hybridization
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Oxidoreductases / antagonists & inhibitors
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Phenotype
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Plasmids
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Saccharomyces cerevisiae / drug effects
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Saccharomyces cerevisiae / genetics*
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Saccharomyces cerevisiae / growth & development
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Sterol 14-Demethylase
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Transformation, Genetic
Substances
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DNA, Fungal
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Cytochrome P-450 Enzyme System
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Oxidoreductases
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Sterol 14-Demethylase
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NADPH-Ferrihemoprotein Reductase
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DNA Restriction Enzymes
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Leucine
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Ketoconazole