Gene Expression in Mycobacterium Tuberculosis

描述(申请人提供)：结核病，人类的最大祸害之一，在当今世界仍然是一个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病的病原体--结核分枝杆菌。全世界每年约有800万新确诊的结核病病例。在那些患有活动性疾病的人中，每年有近200万人死亡。不足为奇的是，1993年，世卫组织宣布结核病为全球卫生紧急情况，就像今天一样。 在过去的12-14年间，在开发有助于分析结核分枝杆菌基因和基因产物的遗传工具方面取得了重大进展。测定了两株结核分枝杆菌全基因组的核苷酸序列。DNA微阵列分析和双向凝胶电泳分析技术提供了关于在特定环境中生长的结核分枝杆菌全球基因表达和蛋白质生产的有价值的信息。我们开发了一种方法来识别结核分枝杆菌在它们青睐的生态环境--人类巨噬细胞中生长时表达的结核分枝杆菌基因。 虽然已经了解了很多，但结核分枝杆菌发病机制的基本方面仍然未知。利用目前可用的遗传、基因组、蛋白质组和分子生物学工具，我们建议(1)表征TrcRS、Rv1626和PrrAB调控系统的调控规律；(2)通过比较结核分枝杆菌野生型H37Rv株和H37Rv株与人巨噬细胞生长过程中表达的特定基因的突变情况，确定特定基因产物在结核杆菌发病中的作用；(3)通过基因芯片和定量RT-PCR(QRT-PCR)分析比较结核杆菌在人外周血单核细胞来源的巨噬细胞和感染小鼠肺中生长的基因表达。

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.