犬瘟热病毒血凝蛋白与SLAM受体在感染中的分子机制研究

项目名称： 犬瘟热病毒血凝蛋白与SLAM受体在感染中的分子机制研究

项目编号： No.31472196

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 水产学、兽医学

项目作者： 黄娟

作者单位： 青岛农业大学

项目金额： 80万元

中文摘要： 犬瘟热病毒血凝蛋白（H）通过利用SLAM受体在病毒感染中发挥重要作用，而SLAM分子在动物体内分布广泛且数量众多，研究H蛋白和SLAM受体相互作用机理对阐明病毒感染机制有重要意义。本项目拟通过反向遗传机制构建SLAM相关位点突变毒株，接种SLAM受体细胞，利用CDV单抗和SLAM单抗对接毒的细胞上吸附病毒粒子和SLAM受体进行定位并与正常病毒做比较，以研究H蛋白在SLAM受体识别和结合中的作用；利用H蛋白和F蛋白在Vero细胞中一过性表达，观察多核体形成情况，以研究和分析H蛋白特定区域对于启动F蛋白的影响；制备SLAM受体蛋白抗体，免疫组化方法研究犬瘟热感染前后SLAM受体变化和分布。通过此研究探寻犬瘟热病毒SLAM受体和病毒蛋白间作用机制，犬瘟热病毒内部蛋白分子间作用机理，病毒感染与SLAM受体关系，感染对受体表达影响。本研究有助于进一步阐明CDV蛋白在病毒感染中的分子机制，也为病毒致病机理研究提供了方法。

中文关键词： 动物传染病；动物病毒；分子机制

英文摘要： The Hemagglutinin(H) potein of Canine Distemper Virus(CDV) plays an improtant role in the infection when the receptor is SLAM moleculer which abounds in animal body, so the reseach on H protein and SLAM receptor gives a great meaning to illutrate the mechanism of virus infection and action. We utilize virus rescue system to obtain CDV with mutation in H gene, somewhere closely related to SLAM receptor, then to infect SLAM receptor cell, then to use CDVmonoclonal antibody and second antibody to combine virus, meanwhile using antibody to combine SLAM receptor, compared and analysed with normal virus infection to study the function of H protein Recognition and combined with SLAM receptor; A H protein and F protein temporal expression vector was constructed and coreansfected in Vero cell which lead to syncytium to inspect weather some specific postion on H protein has effect on trigger Fusion protein; to prepare SLAM protein monoclonal antibody, to find distribution and mutation of SLAM by immunohistochemical method on both healthy and different stages of infection.We get a conclusion of CDV SLAM receptors and viral protein interaction mechanism, a mechanism of internal protein molecules, the relationship between infection and SLAM receptor an effect of receptor expression in infection. It is significant to understand the molecular mechanism of protein CDV infection and the research of methods of attenuated virus in this study.

英文关键词： Veterinary lemology;Animal viruses;Molecular mechanisms