副溶血性弧菌是水产品中重要食源性致病菌，其耐药性呈逐年增强趋势，对水产品安全与公共健康造成极大威胁，明确其耐药机制及安全风险已成为领域基础前沿科学问题。本项目拟以环丙沙星诱导抗性副溶血性弧菌为材料，采用比较蛋白质组学、转录组学等技术筛选副溶血性弧菌耐药膜脂蛋白转运系统Lol关键蛋白及其基因，异源克隆表达Lol关键蛋白，采用高分辨X射线和冷冻电镜技术解析蛋白质三维立体结构，探明Lol系统与耐药膜脂蛋白相互作用的结构域，阐明耐药膜脂蛋白的转运机理和调控机制。同时，运用副溶血性弧菌耐药表型变异系数CVphenotype、耐药微进化变异系数CVmicroevolution及耐药蛋白结构异质性变异系数CVprotein-structure等参数系统修正传统风险评估模型，构建副溶血性弧菌耐药风险评估宏模型。研究成果为靶向破解副溶血性弧菌耐药防御屏障系统提供坚实理论基础，开创细菌耐药风险评估新纪元。

Vibrio parahaemolyticus is the main food-borne pathogen in aquatic products, and its drug resistance has been increasing year by year. It poses a great threat to aquatic product safety and public health. It is clear that its drug resistance mechanism and safety risk have become a fundamental frontier scientific issues. In this project, ciprofloxacin-induced resistance to Vibrio parahaemolyticus was used as a material, and comparative proteomics and transcriptomics sequencing were used to screen key proteins and genes of lipoprotein Lol transport system in Vibrio parahaemolyticus. The heterologous clone expresses the key protein of Lol, and the three-dimensional structure of the protein is analyzed by high-resolution X-ray and cryo-electron microscopy. The domain of the interaction between Lol system and drug-resistant membrane lipoprotein is clarified, and the transport & regulatory mechanism of drug-resistant membrane lipid protein is elucidated. At the same time, the traditional risk assessment model was modified by using the parametric variability coefficient CVphenotype, CVmicroevolution and CVprotein-structure, and then the meta-model for risk assessment of Vibrio parahaemolyticus antibiotic resistance was constructed. The research results will provide a solid theoretical basis for the targeted cracking of Vibrio parahaemolyticus antibiotic resistance defense barrier system, and open up a new era of bacterial antibiotic resistance risk assessment, which is of great strategic significance for safeguarding food safety and maintaining public health.