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通过差异显示法在大鼠口腔念珠菌病模型中检测到白色念珠菌两个基因的上调。

Up-regulation of two Candida albicans genes in the rat model of oral candidiasis detected by differential display.

作者信息

Zhao X J, Newsome J T, Cihlar R L

机构信息

Department of Microbiology and Immunology, Georgetown University, Washington, DC 20007, USA.

出版信息

Microb Pathog. 1998 Sep;25(3):121-9. doi: 10.1006/mpat.1998.0218.

Abstract

Candida albicans is an opportunistic fungal pathogen responsible for the largest percentage of fungal-mediated oral and oesophageal disease. In this regard, knowledge concerning patterns of gene expression during the establishment and/or maintenance of infection may be the key to the design of new strategies for treatment, as well as providing insight into pathogenesis. To address this issue, experiments were performed that utilized differential display to compare the spectrum of C. albicans genes expressed during oral infection versus growth in in vitroculture. Experimentally, the rat model of oral candidiasis served as the in vivo source. After initiation of infection and subsequent harvesting of C. albicans from the rat oral cavity, RNA was isolated, and used with a small number of primers in reverse-transcriptase polymerase chain reaction (RT-PCR) and differential display experiments. Fragments unique to in vivo samples were subcloned and sequenced. Southern blot analysis verified the origin of seven fragments as fromC. albicans. Additionally, specific RT-PCR confirmed that two of these fragments represented genes that were up-regulated during C. albicans in vivo growth in the rat model. Database searches indicated the fragments share homology with a member of the C. albicans agglutinin gene family and to a bacterial gene (gidB) possibly involved in cell division.

摘要

白色念珠菌是一种机会性真菌病原体,在真菌介导的口腔和食管疾病中占比最大。在这方面,了解感染建立和/或维持过程中的基因表达模式可能是设计新治疗策略的关键,同时也有助于深入了解发病机制。为解决这个问题,进行了一些实验,利用差异显示技术比较白色念珠菌在口腔感染期间与体外培养生长时表达的基因谱。实验中,口腔念珠菌病大鼠模型作为体内来源。在感染开始并随后从大鼠口腔采集白色念珠菌后,分离RNA,并在逆转录聚合酶链反应(RT-PCR)和差异显示实验中与少量引物一起使用。体内样品特有的片段被亚克隆并测序。Southern印迹分析证实七个片段来自白色念珠菌。此外,特异性RT-PCR证实其中两个片段代表在大鼠模型中白色念珠菌体内生长期间上调的基因。数据库搜索表明这些片段与白色念珠菌凝集素基因家族的一个成员以及可能参与细胞分裂的细菌基因(gidB)具有同源性。

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