Gene Expression in Mycobacterium Tuberculosis

描述（由申请人提供）：结核病是人类最大的疾病之一，仍然是当今世界严重和重大的健康问题。世界卫生组织估计，世界上三分之一的人口感染了结核分枝杆菌，结核病的病原体。全世界每年诊断出约800万新结核病例。在患有活动性疾病的人中，每年有近200万人死亡。难怪世卫组织在1993年宣布结核病是一个全球卫生紧急情况，今天仍然如此。 在过去的12-14年中，在开发遗传工具以便于分析M.结核对两株M.结核病已确诊。DNA微阵列分析和双向凝胶电泳分析技术提供了有价值的信息，全球基因表达和蛋白质生产的M。结核病在特定环境中生长。我们开发了一种鉴定M.结核病基因，当分枝杆菌生长在他们喜欢的生态位，人类巨噬细胞。 虽然已经了解了很多，M。结核病的发病机制尚不清楚。利用目前可用的一些遗传学、基因组学、蛋白质组学和分子生物学工具，我们建议（1）表征TrcRS、Rv 1626和PrrAB调节系统的调节子，（2）确定特定基因产物对M.结核病发病机制的比较野生型M。结核分枝杆菌H37 Rv和在人巨噬细胞生长过程中表达的特定基因中具有突变的H37 Rv菌株;通过cDNA微阵列和定量RT-PCR（QRT-PCR）分析，研究了结核病在人外周血单核细胞衍生的巨噬细胞和感染的小鼠肺中的生长。

DESCRIPTION (provided by applicant): Tuberculosis, one of the great scourges of humankind, continues to be a serious and significant health problem in the world today. The World Health Organization estimates that one-third of the world's population is infected with Mycobacterium tuberculosis, the causative agent of TB. Approximately 8 million new cases of TB are diagnosed annually worldwide. Of those individuals with active disease, nearly 2 million die each year. Small wonder, then that in 1993, WHO declared tuberculosis to be a global health emergency, as it remains today. During the past 12-14 years, significant progress has been made in the development of genetic tools to facilitate analysis of the genes and gene products of M. tuberculosis. The nucleotide sequences of the entire genomes of two strains of M. tuberculosis have been determined. The techniques of DNA microarray analysis and two-dimensional gel electrophoresis analysis have provided valuable information about global gene expression and protein production by M. tuberculosis grown in defined environments. We developed a method to identify M. tuberculosis genes that are expressed when the mycobacteria are growing in their favored ecological niche, the human macrophage. Although much has been learned, fundamental aspects of M. tuberculosis pathogenesis remain unknown. Employing a number of currently available genetic, genomic, proteomic, and molecular biological tools, we propose to (1) characterize the regulons of the TrcRS, Rv1626, and PrrAB regulatory systems, (2) determine the contribution of specific gene products to M. tuberculosis pathogenesis by comparisons between wild-type M. tuberculosis H37Rv and strains of H37Rv with mutations in specific genes expressed during growth in human macrophages and (3) compare gene expression by M. tuberculosis growing in human peripheral blood monocyte-derived macrophages and in infected mouse lungs by cDNA microarray and quantitative RT-PCR (QRT-PCR) analyses.