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Infection and Immunity, December 2000, p. 6712-6719, Vol. 68, No. 12
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Tetracycline-Regulatable System To Tightly Control Gene Expression in the Pathogenic Fungus Candida albicans

Hironobu Nakayama,* Toshiyuki Mio,dagger Shigehisa Nagahashi,dagger Michiko Kokado,dagger Mikio Arisawa,dagger and Yuko Aokidagger

Department of Mycology, Nippon Roche K. K. Research Center, 200 Kajiwara, Kamakura, Kanagawa 247-8530, Japan

Received 27 January 2000/Returned for modification 10 March 2000/Accepted 13 September 2000

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.

* Corresponding author. Mailing address: Department of Oncology, Nippon Roche K. K. Research Center, 200 Kajiwara, Kamakura, Kanagawa 247-8530, Japan. Phone: 81-467-47-2218. Fax: 81-467-45-6782. E-mail: hironobu.nakayama{at}roche.com.

dagger Present address: Department of Oncology, Nippon Roche K. K. Research Center, 200 Kajiwara, Kamakura, Kanagawa 247-8530, Japan.

Infection and Immunity, December 2000, p. 6712-6719, Vol. 68, No. 12
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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