Isolation of a Cytochrome P-450 Structural Gene From Saccharomyces Cerevisiae

Gene. 1986;45(3):237-45. doi: 10.1016/0378-1119(86)90021-1.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cytochrome P-450 Enzyme System / genetics*
  • DNA, Recombinant / metabolism
  • Drug Resistance
  • Genes*
  • Genes, Fungal*
  • Genetic Vectors
  • Genotype
  • Ketoconazole / pharmacology
  • Plasmids
  • Protein Biosynthesis
  • Saccharomyces cerevisiae / drug effects
  • Saccharomyces cerevisiae / genetics*
  • Sequence Homology, Nucleic Acid

Substances

  • DNA, Recombinant
  • Cytochrome P-450 Enzyme System
  • Ketoconazole

Associated data

  • GENBANK/M15663