猪瘟病毒囊膜蛋白E2的锚定序列部分缺失对其在细胞内表达的影响  

Partial deletion of the membrane anchor sequence of classical swine fever virus envelope glycoprotein E2 decreases its intracellular expression

作　　者：贺番[1] 王靖飞[2] 郝永清[1] 仇华吉[2] 

机构地区：[1]内蒙古农业大学动物科学与医学学院,内蒙古呼和浩特010018 [2]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/猪传染病研究室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2010年第5期342-346,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家973计划项目(2005CB523202)

摘　　要：为了将猪瘟病毒(CFSV)的囊膜蛋白表达在细胞表面,本研究根据与CSFV同属的牛病毒性腹泻病毒(BVDV)的囊膜蛋白E2锚定序列,推测出CSFV的E2锚定序列。利用生物信息学软件对CSFV的E2锚定序列进行分析、建模与部分缺失,构建成4种不同的表达CSFV囊膜蛋白EmsE1E2的片段。将以上4种片段分别连接到真核表达载体pCAGGS中,转染幼仓鼠肾(BHK21)细胞,利用抗E2蛋白的单克隆抗体对表达蛋白进行westernblot检测与间接免疫荧光(IFA)检测,研究结果表明:跨膜区中锚定序列的部分缺失会降低E2在细胞内的表达,为猪瘟伪型病毒的研制提供了实验依据。In this study,effect of partial deletion of a putative anchor sequence on the surface expression of the envelope glycoprotein of classical swine fever virus（CSFV） was investigated.Four DNA fragments expressing envelope glycoprotein ErnsE1 E2 of CSFV were constructed,each containing a VSV signal peptide and a partially deleted membrane anchor sequence similar to the membrane anchor of bovine pestivirus envelope glycoprotein E2.The DNA fragments were cloned into the expression vector pCAGGS and transfected to BHK21 cells.Protein expression was detected by indirect immunofluorescence assay（IFA） and western blot using E2-specific monoclonal antibodies.The results showed that deletion of partial sequences of the membrane anchor decreased the intracellular expression of E2.The study provided references for preparation the pseudotype virus of CSFV.

关 键 词：猪瘟病毒 E2囊膜蛋白 锚定序列 分子模拟技术 

分 类 号：S852.65[农业科学—基础兽医学]