结核分枝杆菌致病基因簇泛基因组偶联定位及功能注释

The virulence genes of Mycobacterium tuberculosis has complex composition and coupling with each other, which caused that the infection ways of pathogen and pathogenic mechanism can not be reflected by studying on the single virulence genes or a single signaling pathway. Furthermore, due to the limitations of experimental technical, it is still impossible for us to establish the Mycobacterium tuberculosis pathogenic genomics and systematical analysis of regulatory networks. . The purpose of this project is to use the visualization of the genome，dynamic simulation of the signal transmission network and high-throughput scanning of central node molecule base on protein-protein interaction to identify virulence genes and simulate the pathogenic pathways in mycobacterium tuberculosis. We also used Tn5 transposon display system build a target molecules mutant library, detect the gene function of each target, analysis their feasibility as central regulatory molecules and node protein of pathogenic gene clusters. Ultimately, we want to build a platform for dynamic network, which could make govern rules and regulatory mechanisms of the pathogen signal pathways more comprehensive and provide a new idea for similar studies of other pathogens..

结核分枝杆菌的致病基因成分复杂、彼此偶联，构成了复杂的基因信号调控网络，孤立地研究单个致病基因或单一信号传导通路往往不能确切地反映病原菌的感染途径及致病机制。由于实验技术的限制尚无法建立结核分枝杆菌致病基因组及相关调控网络的系统解析。. 本项目利用前期建立的基因组的可视化，信号传导网络的动态模拟，调控中枢节点分子的高通量注释及信号传导蛋白偶联等手段实现病原菌致病因子的鉴别及网络调控的建模，并且采用本实验室构建的Tn5转座子展示系统构建靶分子突变库，选择基因组动态映射模型筛选的中枢调控分子及节点蛋白进行功能验证，最终实现结核分枝杆菌致病基因簇的泛基因组偶联定位及功能域注释，从而更全面地描述生物系统的运行规律和调节机制，为其它病原菌的同类研究提供新思路。

该课题基本完成研究目标制定的所有任务。首先通过结核分枝杆菌耐药基因rpoB的突变位点的分析，发现其具有地域性和时间性，并且其突变频率逐年下降；我们又通过文献计量法共找到584个结核分枝杆菌H37Rv的致病基因，又通过分析其pathway及operon网站分析发现了这些致病基因潜在的pathway关联、功能分类及operon关联，并推测出13个极有可能是结核分支杆菌的致病基因，这对治疗结核病以及与结核病相关疾病具有重要意义；我们通过对GEO数据库筛选到的1075个差异基因，并构建了转录因子网络调控图。通过统计分析转录因子及其网络调控图的网络节点，我们推测il6、jun、tp53与转录因子src、tnf、mapk14 一起调控机体的免疫功能，逃避宿主免疫系统的攻击，最终可能在宿主体内能以非增殖状态存在从而造成延缓性病程和导致复发的持续感染。