基于Split-GFP技术的幽门螺杆菌IV型分泌系统效应蛋白鉴定及转运识别机制研究

Type IV secretion system (T4SS) is considered as an important virulent factor of Helicobacter pylori (H. pylori), playing a key role in H. pylori pathogenesis. In recent years, numerous studies have focused on the composition and function of T4SS. However, except CagA, the other effector proteins of T4SS and their translocation process are not clear. In this project, Split-GFP (green fluorescent protein) technology based on the principle of bimolecular fluorescence complementation will be used to study the translocation of H. pylori proteins. Establishment and optimization of method to detect injection of effector proteins through H. pylori T4SS into receptor cells, which will provide an effective tool in H. pylori T4SS study. Based on this method, the hypothetical effector proteins on Cag PAI will be identified. Finally, the recognition sequence of H. pylori T4SS will be further analyzed. These findings would be beneficial to reveal the mechanism of protein translocation by T4SS and pathogenic mechanism of H. pylori infection.

Cag致病岛编码的IV型分泌系统（T4SS）已逐渐成为公认的幽门螺杆菌（H. pylori）重要的毒力因子，与致病、致癌作用紧密相关。大量的研究试图揭示H. pylori T4SS的蛋白组成、结构与功能，然而对其除CagA以外的效应蛋白以及效应蛋白的识别、转运机制还不清楚。本项目将基于双分子荧光互补原理的GFP拆分（Split-GFP）技术应用到H. pylori的蛋白转运研究中，建立并优化Split-GFP技术用于T4SS效应蛋白转运研究的检测方法，为H. pylori T4SS及效应蛋白的深入研究提供有效手段；在此基础上，逐一分析Cag致病岛编码的潜在效应蛋白能否通过T4SS转运到宿主细胞中，以期发掘H. pylori T4SS新的效应蛋白；进一步分析T4SS对效应蛋白的转运识别序列，揭示蛋白转运识别机制，为深入了解H. pylori T4SS的蛋白转运机制及其致病、致癌作用奠定基础。