Isolation of a cytochrome P-450 structural gene from Saccharomyces cerevisiae.
We have transformed a Saccharomyces cerevisiae host with an S. cerevisiae genomic library contained in the shuttle vector YEp24 and screened the resultant transformants for resistance to ketoconazole (Kc), an inhibitor of the cytochrome P-450 (P-450) enzyme lanosterol 14 alpha-demethylase. Two plasmids were isolated which transformed yeast to both increased resistance to Kc and increased levels of total P-450. Hybrid-selection and immunoprecipitation experiments showed that these plasmids, pVK1 and pVK2, contained the structural gene for an S. cerevisiae P-450. This conclusion was confirmed by the nucleotide sequence of a portion of pVK2, which revealed an open reading frame encoding a characteristic P-450 heme-binding region.
D000595:Amino Acid Sequence; D001483:Base Sequence; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D004274:DNA, Recombinant / Q000378:metabolism; D004351:Drug Resistance; D005796:Genes; D005800:Genes, Fungal; D005822:Genetic Vectors; D005838:Genotype; D007654:Ketoconazole / Q000494:pharmacology; D010957:Plasmids; D014176:Protein Biosynthesis; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid
D004274:DNA, Recombinant; D003577:Cytochrome P-450 Enzyme System; D007654:Ketoconazole