类猪圆环病毒P1蛋白的鉴定及其功能的初步分析

项目名称： 类猪圆环病毒P1蛋白的鉴定及其功能的初步分析

项目编号： No.31272574

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 农业科学

项目作者： 温立斌

作者单位： 江苏省农业科学院

项目金额： 80万元

中文摘要： 类猪圆环病毒P1是我们在国际上首次发现并报道、迄今为止拥有最小基因组的动物病毒。前期研究表明P1对仔猪有致病性，是危害养猪业健康发展的又一具有现实意义的病原。由于病毒蛋白在病毒感染发病机制中起着重要的作用，所以，本课题拟通过Northern杂交、RT-PCR和RACE对P1进行转录本分析，通过P1病毒各蛋白的合成肽多克隆抗体进行病毒感染细胞后病毒各蛋白表达分析，对P1病毒蛋白的组成进行鉴定。然后，通过对病毒各蛋白在细胞中的定位，组装病毒样颗粒的能力，蛋白表达缺失对病毒复制、自身其它基因和细胞基因表达谱的影响等，初步研究P1病毒各蛋白的功能。同时将提纯的病毒和无细胞蛋白表达的病毒蛋白进行蛋白质印迹和质谱鉴定，综合对P1病毒蛋白的组成进行鉴定及对蛋白功能进行初步的分析。本课题的完成对于阐明P1病毒感染的发病机制具有重要的意义，同时也为有效控制P1感染的发生与流行提供科学依据和理论基础。

中文关键词： 类猪圆环病毒P1；转录本；病毒样颗粒；信号通路；

英文摘要： We first discovered and reported the porcine circovirus-like P1, a novel animal virus with the smallest genome to date. Previous studies have shown that P1 belongs to the pathogenic virus to piglets and has a practical significance for the swine industry. As viral proteins plays an important role in pathogenesis of a virus infection, identification and preliminary functional analysis of proteins of porcine circovirus-like P1 will be carried out as follows: transcriptional analysis of P1 to identify viral genes and their corresponding mRNA transcripts by northern blot, RT-PCR, and RACE analysis; IHC and/or IFA to detect genes expression of P1 by the polyclonal antibody against synthetic peptides derived from ORFs; The position of the viral proteins in transfected cells; the ability to self-assemble into virus-like particles(VLPs); effects on DNA replication, the other viral gene transcription level, and host gene transcription level by deletion analysis of the vial protein expression; identification of both viruses purified and viral proteins produced by cell-free protein expression by western blot and mass spectrometry. The completion of this project will clarify pathogenesis of P1 infection and provide the scientific basis and theoretical foundation of effectively controlling the incidence and prevalence of P1

英文关键词： porcine circovirus like virus P1；transcript；virus-like particles；signal pathway；