体外共表达的猪繁殖与呼吸综合征病毒GP5和M蛋白可形成异源二聚体  

Co-expressed GP5 and M proteins of porcine reproductive and respiratory syndrome virus can form heterodimers

作　　者：马艳 江云波[1] 肖少波[1] 方六荣[1] 陈焕春[1] 

机构地区：[1]华中农业大学动物医学院动物病毒室,武汉430070

出　　处：《微生物学报》2006年第4期639-643,I0004,共6页Acta Microbiologica Sinica

基　　金：国家自然科学基金(30300257;30571385);国家"973项目"(2005CB523200)~~

摘　　要：为了探讨猪繁殖与呼吸综合征病毒(PRRSV)ORF5基因编码的GP5蛋白和ORF6编码的M蛋白体外共表达特性,分别构建了PRRSV ORF5、ORF6单基因或双基因共表达的真核表达质粒pCI-ORF5、pCI-ORF6和pCI-ORF5/ORF6,转染BHK-21细胞,Western blot检测证实共表达的GP5和M蛋白能够形成异源二聚体。同时,以绿色荧光蛋白(EGFP)和红色荧光蛋白(RFP)为示踪,发现当ORF5-EGFP和ORF6-RFP共表达时,能促进GP5蛋白从内质网向高尔基体转运,提示GP5-M异源二聚体的形成可能与GP5蛋白的翻译后修饰、转运、定位有关。In order to investigate the characterization of in vitro co-expressed GP5 and M proteins of porcine reproductive and respiratory syndrome virus （PRRSV）, eukaryotic expression plasmids pCI-ORF5 （expressing GP5 protein alone）, pCI-ORF6 （expressing M protein alone）, and pCI-ORFS/ORF6 （co-expressing GP5 and M proteins） were constructed. After transient transfection, Western blot analysis under nonreducing condition demonstrated that co-expressed GP5 and M proteins could form disulfide-linked heterodimers （GPS-M） in transiently transfected BHK-21 cells. To further study the influence of GPS-M heterodimers formation on the subcellular Iocalizations of GP5 or M proteins, green fluorescence protein （EGFP） and red fluorescence protein （RFP） were used as markers. The results of fluorescence distribution showed that co-expressed GPS-EGFP chimera and M-RFP chimera boosted the transport of GP5 from the endoplasmic reticulum （ER） to the Golgi complex, indicating that the formation of GPS-M heterodimers may be involved in posttranslational modification, transport, and subcellular localization of GPS. These results presented here lay foundation to further study the molecular mechanism of GPS-M heterodimer formation and its role in protective immunity of PRRSV.

关 键 词：PRRSV ORF5 ORF6 双基因共表达 异源二聚体 

分 类 号：S852.65[农业科学—基础兽医学]