结核分枝杆菌cdigmp代谢相关四蛋白的生物化学研究.docxVIP

ABSTRACTCountless ABSTRACT Countless millions of people in the world have died from tuberculosis，a chronic infectious disease caused by the tubercle bacillus．The complete genome sequence of the best-characterized Mycobacterium tuberculosis strain，H37Rv,has been determined and analysed in order tO improve the understanding of the biology of this slow-growing pathogen and to help finding new prophylactic and therapeutic interventions． Bis一(3’一5’)-cyclic dimeric guanosine monophosphate(C-di-GMP)has come to the limelight as one of the result ofthe recent advances in microbial genomics．C．di．GMP is known嬲a novel global second messenger in bacteria．The GGDEF and EAL domain proteins，which are involved in C-di—GMP synthesis and degradation, respectively,are ubiquitous in bacterial genomes．These proteins affect cell differentiation and multiceUular behaviour as well as the interactions between the microorganisms and their eukaryotic hosts． In this study,we cloned and purified the Mycobacterium tuberculosis protein Rv 1 3 54c，which contains both GGDEF domain and EAL domain．In the Rdar experiment and enzymatic assay,we confirmed it could synthesize C-di-GMP from GTP and degrade GTP t0 GDP．By the enzymatic assay of the different domains of Rvl 354c，we found the GA莹domain is requied for the activity of GGDEF domain and the EAL domain call degrade GTP tO GDP by itself．The binding asssy confirmed the GAF domain Call bind hemachrome but not cAM限 Key Words：tuberculosis，Mycobacterium tuberculosis，Bis·(3’·5’)一cyclic dimeric guanosine monophosphate， GGDEF, EAL， Rvl 354c， GAF, hemachrome IV 中国科学技术大学学位论文原创性声明本人声明所呈交的学位论文，是本人在导师指导下进行研究工作所取得的成 中国科学技术大学学位论文原创性声明 本人声明所呈交的学位论文，是本人在导师指导下进行研究工作所取得的成 果。除已特别加以标注和致谢的地方外，论文中不包含任何他人已经发表或撰写 过的研究成果。与我一同工作的同志对本研究所做的贡献均已在论文中作了明确 的说明。 作者签名：』麟 签字日期：．垒!!：兰 中国科学技术大学学位论文授权使用声明 作为申请学位的条件之一，学位论文著作权拥有者授权中国科学技术大学拥 有学位论文的部分使用权，ep．学校有权按有关规定向国家有关部门或机构送交 论文的复印件和电子版，允许论文被查阅和借阅，可以将学位论文编入有关数据 库进行检索，可以采用影印、缩印或扫描等复制手段保存、汇编学位论文。本人 提交的电子文档的内容和纸质论文的内容相一致。 保密的学位论文在解密后也遵守此规定。