Analysis of a Candida albicans gene that encodes a novel mechanism for resistance to benomyl and methotrexate
| Autor: | Jan Kopf, Jessica A. Gorman, Mary E. Fling, Yigal Koltin, Aviva Tamarkin, Herbert A. Smith |
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| Rok vydání: | 1991 |
| Předmět: |
Genes
Fungal Molecular Sequence Data Restriction Mapping Saccharomyces cerevisiae Polymerase Chain Reaction Open Reading Frames chemistry.chemical_compound Plasmid Restriction map Candida albicans Dihydrofolate reductase Escherichia coli Genetics Amino Acid Sequence Codon Site-directed mutagenesis Molecular Biology Base Sequence biology Nucleic Acid Hybridization Benomyl Drug Resistance Microbial biology.organism_classification Molecular biology Dihydrofolate reductase inhibitor Corpus albicans Methotrexate chemistry Mutagenesis Site-Directed biology.protein |
| Zdroj: | Molecular and General Genetics MGG. 227:318-329 |
| ISSN: | 1432-1874 0026-8925 |
| DOI: | 10.1007/bf00259685 |
| Popis: | The pathogenic yeast, Candida albicans, is insensitive to the anti-mitotic drug, benomyl, and to the dihydrofolate reductase inhibitor, methotrexate. Genes responsible for the intrinsic drug resistance were sought by transforming Saccharomyces cerevisiae, a yeast sensitive to both drugs, with genomic C. albicans libraries and screening on benomyl or methotrexate. Restriction analysis of plasmids isolated from benomyl- and methotrexate-resistant colonies indicated that both phenotypes were encoded by the same DNA fragment. Sequence analysis showed that the fragments were nearly identical and contained a long open reading frame of 1694 bp (ORF1) and a small ORF of 446 bp (ORF2) within ORF1 on the opposite strand. By site-directed mutagenesis, it was shown that ORF1 encoded both phenotypes. The protein had no sequence similarity to any known proteins, including beta-tubulin, dihydrofolate reductase, and the P-glycoprotein of the multi-drug resistance family. The resistance gene was detected in several C. albicans strains and in C. stellatoidea by DNA hybridization and by the polymerase chain reaction. |
| Databáze: | OpenAIRE |
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