结核分枝杆菌中转录因子介导的耐药调控机制研究

项目名称： 结核分枝杆菌中转录因子介导的耐药调控机制研究

项目编号： No.81471996

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 何正国

作者单位： 华中农业大学

项目金额： 72万元

中文摘要： 结核病全球蔓延，细菌耐药问题十分严重。然而，尽管目前已经清楚药物靶标基因突变是导致临床菌株耐药的主要原因，但是有关结核分枝杆菌耐药性形成的其它调控机制还知之甚少，相关的调控因子也还未系统地鉴定。为此，本项目拟首先大规模克隆结核分枝杆菌的转录调控基因，构建基因超量表达文库，系统筛选对结核分枝杆菌药物抗性能力有显著影响的转录因子。在此基础上，进一步采用EMSA、ChIP、转录组学和蛋白质组学分析以及基因敲除等方法，综合鉴定这些转录因子蛋白与它们的靶启动子DNA之间的相互作用，发现和剖析相关信号通路，从而系统阐明结核分枝杆菌中转录因子介导的细菌耐药调控新机制。本项目的实施将能显著提升我们对基因转录调控导致结核分枝杆菌耐药性形成的分子机制的认识，有助于推动新药靶标的发掘，为结核病的防控提供新思路。

中文关键词： 耐药机制；基因突变；结核分枝杆菌；多重耐药；细菌耐药性

英文摘要： Tuberculosis (TB) has been a menace to human health throughout the world. The drug resistance of Mycobacterium tuberculosis has become an extremely serious problem to any attempt to control TB. It is basically known that acquired resistance to the first-line antibiotics because of sequential accumulation of mutations in target genes has resulted in the emergence of drug resistant TB. However, the additional regulating mechanisms on the drug restance of M. tuberculosis are largely unclear and the involved transcription factor remains to be identified. In this proposal, we will firstly clone almost entire regulatory genes from the genome of M. tuberculosis into overexpression plasmids and screen potential regulators which can obviously affect the ability of the mycobacterial drug resistance. Then, using multiple methods, including EMSA, ChIP, transcriptomics, proteomics and gene knock-out assays, we will comprehensively characterize the interaction between the regulator and the target promoter DNA, and discover new signaling pathways in the pathogen. Finally, we will explore on the relationships betwen the regulator-involved signaling pathways and mycobacterial drug-resistance. Therefore, this study would significantly improve our understanding on the transcription regulatory mechanisms of the bacterial drug-resistance in M. tuberculosis. This also will definitely help explore new drug target and develop ideal stretgy for finally controling TB in the future.

英文关键词： Mechanism of drug resistance;Gene mutation;M.tuberculosis;Multi-drug resistance;Bacterial drug resistance