Tetracycline-Regulatable System To Tightly Control Gene Expression in the Pathogenic Fungus Candida albicans
AUTOR(ES)
Nakayama, Hironobu
FONTE
American Society for Microbiology
RESUMO
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.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=97771Documentos Relacionados
- An extrachromosomal tetracycline-regulatable system for mammalian cells
- Inducible Overexpression of a Toxic Protein by an Adenovirus Vector with a Tetracycline-Regulatable Expression Cassette†
- Candida Albicans: The Pathogenic Fungus
- Structure and regulation of the HSP90 gene from the pathogenic fungus Candida albicans.
- Structure and regulation of the Hsp90 gene from the pathogenic fungus Candida albicans.