Expression of a proteolipid gene from a high-copy-number plasmid confers trifluoperazine resistance to Saccharomyces cerevisiae
Autor: | J. Kwong, N. Neff, Cheng-Kon Shih, E. Montalvo |
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Rok vydání: | 1990 |
Předmět: |
Protein subunit
Proteolipids Saccharomyces cerevisiae Genes Fungal Molecular Sequence Data Restriction Mapping Gene Expression Biology Haploidy medicine.disease_cause chemistry.chemical_compound Plasmid Sequence Homology Nucleic Acid Gene expression medicine Escherichia coli Animals Amino Acid Sequence DNA Fungal Gene Molecular Biology Base Sequence Nucleic Acid Hybridization Drug Resistance Microbial Cell Biology biology.organism_classification Molecular biology Yeast Trifluoperazine Biochemistry chemistry Cattle Chromosome Deletion Oligonucleotide Probes DNA Research Article Plasmids |
Zdroj: | Molecular and Cellular Biology. 10:3397-3404 |
ISSN: | 1098-5549 0270-7306 |
Popis: | A wild-type haploid yeast strain was transformed with a library of wild-type yeast DNA fragments ligated into a high-copy-number plasmid vector (YEp24). The pooled URA+ transformants were plated on rich medium containing a lethal concentration of trifluoperazine (TFP). Plasmids rescued into Escherichia coli from TFP-resistant yeast colonies contained overlapping DNA fragments from a unique region of yeast chromosome XVI. Deletion and disruption experiments, mini-Tn10 LUK hop analysis, and DNA sequencing defined a novel gene with significant amino acid identity to bovine and yeast vacuoletype proteolipid subunits. This is the second locus identified that can be altered to confer TFP resistance to Saccharomyces cerevisiae and that has significant amino acid identity to a vacuolar ATPase subunit. This suggests that a target for TFP in S. cerevisiae is the electrogenic membranes of the vacuolar network and that alteration of expression or activity of vacuolar proton ATPase subunits is a general mechanism for TFP resistance in this yeast. |
Databáze: | OpenAIRE |
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