in vivo searching for essential genes in Candida glabata

念珠菌属中的几个种类可在免疫功能低下的宿主中引起全身性感染，例如感染艾滋病毒或接受器官移植或正在接受强化化疗的患者。全基因组分析已经开始，以便系统地了解这些真菌和促进新的抗真菌药物的开发。白色念珠菌是念珠菌属中最常见和最严重的病原体，其次是光滑念珠菌。近年来，由于对抗真菌药物的敏感性较低，感染光棘球蚴的患者呈上升趋势。酿酒酵母（c.g allabrata）与酿酒酵母菌（Saccharomyces cerevisiae）有密切的亲缘关系，为酿酒酵母菌的基因功能分析提供了分子工具和数据库。基于这些原因，我们一直在研究C.glabrata。2004年，我们构建了青叶蝉基因组文库。确定了它们的序列，并分配了启动子区域。基因操作是通过将启动子区域替换为可调节的启动子来进行的，该启动子可以在体内和体外工作。2005年，用含有70个与靶基因同源序列的长低聚dna扩增了可调节启动子区域。将DNA盒插入到光斑田鼠的受体细胞中。分别为183个基因构建了启动子替代菌株。利用这些菌株，进一步进行了体内生长试验。

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.