抑制素2介导克氏原螯虾应对白斑病毒感染的分子机制研究

项目名称： 抑制素2介导克氏原螯虾应对白斑病毒感染的分子机制研究

项目编号： No.31502199

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 农业科学

项目作者： 兰江风

作者单位： 华中农业大学

项目金额： 20万元

中文摘要： 白斑病的爆发每年给克氏原螯虾养殖造成巨大的损失。筛选有效的抗病毒分子是白斑病防治的重要途径。抑制素（Prohibitins, PHBs）在宿主感染病毒免疫反应中起到重要作用，但分子机制还有待揭示。申请人研究发现：病毒刺激后，克氏原螯虾PHB2体内转录水平明显增加。以此为基础，本研究拟通过体内/体外干扰和过表达PHB2的方式，理清PHB2与病毒复制的关系，阐明PHB2对信号转导与转录激活因子（STAT）表达水平和磷酸化水平的影响，并筛选PHB2调控的免疫效应分子；利用Pull-down、CoIP和免疫荧光共定位等技术，研究PHB2与病毒蛋白及STAT的相互作用。本课题将系统研究PHB2在宿主感染病毒免疫反应中的功能，最终确定PHB2同病毒侵染与复制的关系，明确PHB2在Jak/STAT信号调控中的功能。本研究将增补先天免疫知识体系，同时为白斑病防治提供新的思路。

中文关键词： 白斑综合症病毒；克氏原螯虾；抑制素；先天免疫；信号通路

英文摘要： White spot syndrome virus (WSSV) infection has caused great economic loss in Procambarus clarkii culture. Screening the antiviral molecules is an effective way to prevent the WSSV infection. Prohibitins (PHBs) play an important role in host immune responses to many viruses infection, but the molecular mechanism of PHBs response to WSSV infection remains to be revealed. We found that the expression 　of the PHB2- mRNA level was up-regulated significantly after challenge with WSSV. In this proposal, we will measure the replication level of WSSV and the expression and phosphorylation levels of STAT after suppression and over-exppression of PHB2 in vito and in vivo. Subsequently, we will identity the effective molecules regulated by PHB2. The interactions between PHB2 and the viral proteins as well as STAT will be further characterized by pull-down, co-immunoprecipitation and immunofluorescence. Eventually, we will clarify the relationship between PHB2 and the infection and replication of WSSV in crayfish. This study will shed a new light on the innate immune responses of crayfish during the infection of WSSV, and will pave a new way for the development of effective prevention strategies for WSSV infection.

英文关键词： White spot syndrome virus;Procambarus clarkii;Prohibitin;Innate immunity;Signal pathways