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 cells were first transformed with the plasmid carrying the tetracycline repressor-transactivator fusion then with the DNA fragment containing the controllable cassette, the tetracycline operator chimeric promoter The peptide elongation factor 3 and DNA topoisomerase II genes from were cloned and their expression assessed using this system. When the promoter of or was replaced with doxycycline almost completely repressed the expression of both mRNAs, and impaired growth. Repression of the or gene by doxycycline also hampered the survival of cells in mice; in mouse kidneys the number of cells, in which the or promoter was replaced with the 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.


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