利用转座子随机插入文库测序（Tn-Seq）的方法研究霍乱弧菌的耐盐机制并鉴定耐盐相关基因

霍乱弧菌是一种能引起严重腹泻的肠道致病菌，经常在河口水或一些高盐食物中存在并经口感染人类。水体和食物中的高盐压力以及肠道中的高渗环境，是霍乱弧菌必须要应对的环境压力之一。对霍乱弧菌的耐高盐机制进行研究，可以加深我们对细菌应对高盐反应机制的认识，而且利于我们进一步完善对霍乱弧菌生存、传播和致病能力的认识。目前对霍乱弧菌的耐盐机制研究还比较少，本研究拟对霍乱弧菌转座子随机插入文库在正常环境和高盐环境下培养不同时间点的全基因组进行高通量测序，分析其差异，以期在全基因水平上对霍乱弧菌的耐盐机制进行动态性时序性的分析；并对外膜蛋白基因ompT基因和丰度下调且同转运相关的基因进行基因缺失和功能验证；最后对筛选到的基因进行联合缺失，观察它们在霍乱弧菌的耐盐机制中是否起协同效应，以期初步构建一个转运蛋白在霍乱弧菌耐盐中作用的简图。

Vibrio cholera is a serious human intestinal pathogen, often resides and thrives in estuaries and high-salt food, which will encounter many types of high salt stress during its life cycle, including hyperosmotic stress in the intestine and high salt stress in water and food. The ability to tolerate high salt conditions is critical for their survival in the environment and pathogenesis in humans. In this research, we will use transposon insertion site sequencing (often referred to as Tn-seq) to explore the responses and the possible time-course mechanisms of V. cholerae to elevated salt stress. In addition, we further study the ompT gene and the transport genes down-regulated by gene deletion and functional verification to find the new salt-related genes. Finally, we will joint delete the identified salt-related genes to identify whether they have synergistic effects to enhance the salinity tolerance of V. cholera.