Isolation of a cytochrome P-450 structural gene from Saccharomyces cerevisiae

Authors

V. F. Kalb, Molecular Genetics UniversityFollow
J. C. Loper, Molecular Genetics UniversityFollow
C. R. Dey, Molecular Genetics University
C. W. Woods, Molecular Genetics University

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 14α-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. © 1986.

Publication Title

Gene

Recommended Citation

Kalb, V., Loper, J., Dey, C., & Woods, C. (1986). Isolation of a cytochrome P-450 structural gene from Saccharomyces cerevisiae. Gene, 45 (3), 237-245. https://doi.org/10.1016/0378-1119(86)90021-1