Resistance to Imidazoles and Triazoles in Saccharomyces cerevisiae as a New Dominant Marker

• Published in: Plasmid Vol. 30; no. 3; pp. 224 - 233
• Main Authors: Doignon, François, Aigle, Michel, Ribereau-Gayon, Pascal
• Format: Journal Article
• Language: English
• Published: San Diego, CA Elsevier Inc 01.11.1993; Elsevier
• Subjects: Antifungal Agents - toxicity; Base Sequence; Biological and medical sciences; Cytochrome P-450 Enzyme System - biosynthesis; Cytochrome P-450 Enzyme System - genetics; Drug Resistance, Microbial - genetics; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Gene Expression; Imidazoles - toxicity; Microbiology; Miscellaneous; Molecular Sequence Data; Mutagenesis; Mycology; Oligonucleotide Probes; Oxidoreductases - biosynthesis; Oxidoreductases - genetics; Phosphoglycerate Kinase - genetics; Plasmids; Promoter Regions, Genetic; Restriction Mapping; Saccharomyces cerevisiae; Saccharomyces cerevisiae - drug effects; Saccharomyces cerevisiae - genetics; Silanes - toxicity; Sterol 14-Demethylase; Triazoles - toxicity; Triazole; Yeast; Enzyme; Transcription promoter; Transferases; Cytochrome P450; Gene overexpression; Phosphoglycerate kinase; Fungi; Resistance; Chemical modification; Enzymatic activity; Imidazole; Ascomycetes; Saccharomyces cerevisiae; Hybrid gene; Thallophyta
• ISSN: 0147-619X; 1095-9890
• DOI: 10.1006/plas.1993.1054

The imidazole and triazole fungicides inhibit cytochrome P450 14α-lanosterol demethylase (P450 14DM) implicated in the ergosterol biosynthesis pathway, which is specific to fungi and yeasts. Two plasmids were obtained which allow triazole and imidazole resistance in Saccharomyces cerevisiae . The low copy number plasmid (pFD 383) encodes cytochrome P450 C14α lanosterol demethylase under the control of phospho-glycerate-kinase promoter. S. cerevisiae transformed by the pFD 383 plasmid are resistant to imidazoles and triazoles. Moreover, this transformed strain shows increased levels of P450 14DM mRNA and of cytochrome P450. A second low copy number plasmid (pFD 384) carries a mutant cytochrome P450 14α lanosterol demethylase gene, which increases imidazole and triazole resistance. These constructions can be used on a dominant selection marker to transform wild-type yeasts and to confer imidazole and triazole resistance in industrial fermentation.