in vivo searching for essential genes in Candida glabata

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Several species within the genus Candida cause systemic infections in immunocompromised hosts, such as patients with HIV infection or who have undergone organ transplants, or are undergoing intensive chemotherapy. Genome wide analyses have begun in order to systematically understand these fungi and to promote development of new anti-fungal medicines. C.albicans is the most common and most severe pathogen within the genus Candida, followed by C.glabrata as the second most common and serious pathogen. Recently patients who infected with C.glabrata has been increasing, because of low susceptibility against anti-fungal medicine. C.glabrata is a closely related to Saccharomyces cerevisiae, moleculer tools and data bases which are developed for S.cerevisiae, are useful for the analysis of gene function in C.glabrata. For these reason, we have been working on C.glabrata. In 2004, we constructed genomic library of C.glabrata. determined their sequence, and assigned promoter region. The gene manipulation is performed via the promoter region replacement to a reguiatable promoter, which can work in vivo as well as in vitro. In 2005, the regulatable promoter region was amplified with long oligo-DNAs including 70 mer homologous sequence to a target gene. The DNA cassette inserted into the recipient cells of C.glabrata. The promoter replacement strains have been constructed for 183 genes individually. Using these strains, in vivo growth test is carried out further.