Application of differential display-PCR technique in fluconazole-resistance gene expression of Candida.
- Author:
Yu-ning ZHU
1
;
Shi-ming LU
Author Information
- Publication Type:Journal Article
- MeSH: Antifungal Agents; pharmacology; Candida albicans; drug effects; genetics; Drug Resistance, Fungal; genetics; Fluconazole; pharmacology; Fungal Proteins; genetics; Membrane Transport Proteins; genetics; Oxidoreductases; genetics; Polymerase Chain Reaction; methods
- From: Journal of Zhejiang University. Medical sciences 2005;34(2):157-162
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the application of differential display-2PCR(DD-PCR) in research on gene expression of Candida.
METHODSResistance to fluconazole was induced in a Candida albicans isolate 435 from vagina by culturing in YEPD broth with increasing fluconazole concentration in vitro, and the resistant isolate 435-2 (MIC=128 microg/ml ) was obtained after 80 days of incubation. Comparisons between 435 and 435-2 either in fluconazole-containing medium or in drug-free medium were performed with the modified DD-PCR including amplification with long primers, silver staining, reverse dot blot and non-radiographic labeling techniques.
RESULTSThree differential displayed bands were found which showed high homology to alcohol dehydrogenase 1 (ADH1), TOP2 and CDR1, respectively. The up-regulating expression of ADH1 and CDR1 associated with fluconazole resistance was further identified by RT-PCR.
CONCLUSIONThe up-regulating expression of ADH1 and CDR1 was associated with fluconazole resistance in Candida albicans, ADH1 might be a candidate of novel fluconazole resistant gene.
