基于萌发受体GerA的Bacillus subtilis芽孢萌发信号传导机制研究

芽孢是细菌在环境营养缺乏条件下形成的休眠体，具有极低的代谢活性和极强的抗逆性。休眠的芽孢能感知外界环境，一旦环境条件合适，就会萌发生长恢复代谢活动，从而对食品安全和人类健康造成重大威胁。由于萌发是芽孢复苏必须经历的过程，因此控制芽孢萌发是控制芽孢的有效途径。然而，目前对于芽孢萌发过程中的信号传导机制并不清楚。本项目拟以Bacillus subtilis芽孢萌发的经典途径—基于萌发受体GerA的萌发—为研究对象，通过磷酸化蛋白组学、基于TurboID的邻近蛋白组学以及免疫共沉淀等技术寻找GerA的下游蛋白，并结合双分子荧光互补（BiFC）、蛋白体外表达和time-lapse荧光显微镜等技术深入探究GerA与下游蛋白之间的相互作用机制，从而揭示芽孢萌发过程中信号传导机制。本研究将有助于深入理解芽孢萌发过程，为食品工业中更精确有效的控制芽孢奠定理论基础。

Bacteria can form spores upon starvation and spores are in quite a low metabolic level and extremely resistant to adverse conditions. Dormant spores can sense external environment. Once the environment becomes suitable, spores are rapidly revival through germination and outgrowth. Germinated spores may lead to disease or food spoilage, which poses spores a potential risk to food safety and human health. As germination is an inevitable process for spore revival, controlling spore germination is an efficient strategy to control spores. However, the mechanism of signal transduction during spore germination is still not clear. In this study, we are trying to explore the downstream proteins of GerA during spore germination of Bacillus subtilis by phosphorylated proteomics, TurboID catalyzed proximate proteomics and co-immunoprecipitation methods. Moreover, the mechanism of interaction between GerA and downstream protein will further investigated by bimolecular fluorescent complimentary (BiFC), protein express in vitro and time-lapse fluorescent microscopy technologies. These studies will elucidate the mechanism of signal transduction during spore germination, and further our understanding of the germination. This research will provide theoretical foundation for more precisely and efficiently control of spores in food industry.