常见果蔬中诺如病毒吸附受体的解析及其分布规律

项目名称： 常见果蔬中诺如病毒吸附受体的解析及其分布规律

项目编号： No.31301475

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 农业科学

项目作者： 李茜茜

作者单位： 上海应用技术学院

项目金额： 23万元

中文摘要： 诺如病毒是非细菌性流行性腹泻的最主要病原之一，可污染果蔬类农产品，且在加工处理过程中极易导致交叉污染。目前，果蔬中诺如病毒的吸附受体及其分布规律还知之甚少。本项目拟以冰晶核蛋白InaQ作为载体，通过构建InaQ的N末端缺失突变体库，利用膜定位分析方法，获得该蛋白的膜结合核心结构域；以该结构域为载体蛋白，以诺如病毒衣壳蛋白（VP1）为乘客蛋白，以大肠杆菌为受体菌，构建VP1全细胞表面展示体系。以展示体系作为诺如病毒的替代物，收集与受体结合的表面展示重组菌，利用双向电泳和质谱分析等技术，解析并验证果蔬中诺如病毒吸附受体。同时，在多种常见果蔬样本中进行诺如病毒吸附受体定位分析，考查该病毒吸附受体在果蔬中的分布规律。本课题的实施将为果蔬类农产品中诺如病毒的检测方案和污染机制提供理论依据和技术支撑。

中文关键词： 人源诺如病毒；受体；分布；细胞表面展示；生菜

英文摘要： Norovirus is one of the major pathogens in the outbreaks of non-bacterial epidemic diarrhea. Norovirus can be detected from vegetables and fruits, which cause cross-contamination during the processing and handling procedures. However, until now, little is known about the receptors of norovirus and its distributions in those products. In this study, an Ice nucleation protein, named InaQ, will be used as a vehicle. The core membrane binding domain of InaQ will be determined by the deletion mutant library of the N- terminal domain. The membrane localization of this domain will be analyzed. Then, a cell surface display system will be constructed using membrane binding domain as a carrier protein, the main capsid protein (VP1) of norovirus as passenger protein and Escherichia coli as the recipient strain. The constructed cell surface display system will be used as a surrogate to collect the receptors of norovirus from the vegetables and fruits. The receptors will be evaluated by 2-D electrophoresis, mass spectrometry, and so on. Furthermore, the VP1 binding sites will be investigated to figure out the distribution of this virus's receptors in vegetables and fruits. The results from this study will offer theoretical basis and technical supports to the researches of the detection methods and contamination mechanisms of

英文关键词： Human norovirus；receptor；distribution；cell surface display；romaine lettuce