A controllable gene-expression system for the pathogenic fungus Candida glabrata

被引:40
作者
Nakayama, H [1 ]
Izuta, M [1 ]
Nagahashi, S [1 ]
Sihta, EY [1 ]
Sato, Y [1 ]
Yamazaki, T [1 ]
Arisawa, M [1 ]
Kitada, K [1 ]
机构
[1] Nippon Roche Res Ctr, Dept Mycol, Kamakura, Kanagawa 247, Japan
来源
MICROBIOLOGY-UK | 1998年 / 144卷
关键词
Candida glabrata; gene expression; tetracycline-responsive element; peptide elongation factor 3; DNA topoisomerase II;
D O I
10.1099/00221287-144-9-2407
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
A system for controlling gene expression was established in the pathogenic fungus Candida glabrata to elucidate the physiological functions of genes. To control the expression of the gene of interest, the C. glabrata cells were first transformed with the plasmid carrying the tetracycline repressor-transactivator fusion tetR::GAL4, then with the DNA fragment containing the controllable cassette, the tetracycline operator chimeric promoter (tetO::ScHOP1). The peptide elongation factor 3 (CgTEF3) and DNA topoisomerase II (CgTOP2) genes from C. glabrata were cloned and their expression assessed using this system. When the promoter of CgTEF3 or CgTOP2 was replaced with tetO::ScHOP1, doxycycline almost completely repressed the expression of both mRNAs, and impaired growth. Repression of the TOP2 or TEF3 gene by doxycycline also hampered the survival of C. glabrata cells in mice; in mouse kidneys the number of C. glabrata cells, in which the TOP2 or TEF3 promoter was replaced with the tetO::ScHOP1 controllable cassette, did not increase when the mice were given doxycycline. Thus, it appears that the gene repression mediated by doxycycline occurred not only in culture media but also in animals; therefore, this system can be used to elucidate the function of the gene in fungal infections and pathogenesis.
引用
收藏
页码:2407 / 2415
页数:9
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