Tetracycline-regulatable system to tightly control gene expression in the pathogenic fungus Candida albicans.
Conventional tools for elucidating gene function are relatively scarce in Candida albicans, the most prevalent human fungal pathogen. To this end, we developed a convenient system to control gene expression in C. albicans by the tetracycline-regulatable (TR) promoters. When the sea pansy Renilla reniformis luciferase gene (RLUC1) was placed under the control of this system, doxycycline (DOX) inhibited the luciferase activity almost completely. In the absence of DOX, the RLUC1 gene was induced to express luciferase at a level 400- to 1,000-fold higher than that in the presence of DOX. The same results were obtained in hypha-forming cells. The replacement of N-myristoyltransferase or translation elongation factor 3 promoters with TR promoters conferred a DOX-dependent growth defect in culture media. Furthermore, all the mice infected with these mutants, which are still virulent, survived following DOX administration. Consistently, we observed that the number of these mutant cells recovered from the mouse kidneys was significantly reduced following DOX administration. Thus, this system is useful for investigating gene functions, since this system is able to function in both in vitro and in vivo settings.
Nakayama
H
H
Department of Mycology, Nippon Roche K. K. Research Center, 200 Kajiwara, Kamakura, Kanagawa 247-8530, Japan. hironobu.nakayama@roche.com
Mio
T
T
Nagahashi
S
S
Kokado
M
M
Arisawa
M
M
Aoki
Y
Y
D000818:Animals; D002176:Candida albicans / Q000235:genetics*; D004318:Doxycycline / Q000494:pharmacology; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects*; D008297:Male; D051379:Mice; D011401:Promoter Regions, Genetic; D013752:Tetracycline / Q000494:pharmacology*; D015534:Trans-Activators / Q000502:physiology
D015534:Trans-Activators; D013752:Tetracycline; D004318:Doxycycline