Application of differential display-PCR technique in fluconazole-resistance gene expression of Candida / 浙江大学学报·医学版
Journal of Zhejiang University. Medical sciences
;
(6): 157-162, 2005.
Artículo
en Chino
| WPRIM
| ID: wpr-353227
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the application of differential display-2PCR(DD-PCR) in research on gene expression of Candida.</p><p><b>METHODS</b>Resistance 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.</p><p><b>RESULTS</b>Three 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.</p><p><b>CONCLUSION</b>The 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.</p>
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Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Oxidorreductasas
/
Proteínas de Transporte de Membrana
/
Farmacología
/
Candida albicans
/
Proteínas Fúngicas
/
Fluconazol
/
Reacción en Cadena de la Polimerasa
/
Farmacorresistencia Fúngica
/
Genética
/
Métodos
Idioma:
Chino
Revista:
Journal of Zhejiang University. Medical sciences
Año:
2005
Tipo del documento:
Artículo
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