空肠弯曲菌鞭毛合成新转录因子FlhF调控网络解析

空肠弯曲菌是世界范围内主要的人兽共患病病原菌之一，鞭毛及其介导的运动性是其主要致病因素；FlhF是新发现影响鞭毛合成的关键调控因子，现有研究显示其对鞭毛合成及定位具有重要影响，调控机制仍未阐明；前期课题组通过EMSA初步发现该基因可能作为新型转录因子，更直接显著调控鞭毛基因合成，但该功能未见报道。本项目以FlhF新型转录功能为突破点，构建系列缺失株，以启动子活性测定等方法解析其生物学表型；通过ChIP、DNase I足迹检测、CRISPR及系列蛋白构建表达等技术手段，筛选启动子并探究其结合位点，解析相关启动子对报告基因抑制/激活效应；采用BiFC、Co-IP、Pull Down等技术筛选互作蛋白，解析转录偶联机制，丰富并完善其对鞭毛合成的转录调控网络。该项目的开展将以全新视角解析空肠弯曲菌鞭毛基因转录调控机制，为该菌防控感染提供新思路。

Campylobacter jejuni is one of the major zoonotic pathogens in the world. Flagella and its motility are the main causative agents. FlhF is a newly discovered key regulator of flagella synthesis. Previous studies have shown that it has an important impact on the synthesis and localization of flagella, but the regulatory mechanism has not been elucidated. Our group preliminary found by EMSA that the gene may act as a novel transcription factor, but also significantly directly regulate flagellar gene synthesis. However, the function and mechanism have not been reported. The project aims at FlhF novel transcriptional regulation , constructing a series of deletional strains, and their biological phenotypes were analyzed by promoter activity assay or anything. Screening promoters and probing for its binding site by ChIP, DNase I footprinting, CRISPR or anything, followed by analyzing the effects of related promoters on reporter gene to further analysis of FlhF regulatory network in Campylobacter jejuni flagellar synthesis. Finally, screening the interacting proteins by BiFC, Co-IP and Pull Down to parse the transcriptional coupling mechanism, which enrich and perfect the transcription regulatory network of flagella synthesis. The project aims at resolving regulatory network of a novel transcription factor FlhF in Campylobacter jejuni flagella synthesis, clarifying the mechanism of key virulence regulation and providing new ideas for preventing and controlling the infection of the bacterium.