猪繁殖与呼吸综合征病毒GP5蛋白分段表达及其生物学功能的鉴定

Expression and characterization of porcine reproductive and respiratory syndrome virus GP5 fragments

为进一步研究PRRSV GP5蛋白的生物学功能,本研究将已分离的PRRSV北美洲型野毒株TA-12的GP5基因分为4段(79bp～207bp、133bp～330bp、229bp～492bp和382bp～603bp)分别克隆于pGEX-6p-1中,并转化E.coli BL21(Rosetta)细胞进行诱导表达。表达蛋白经纯化后,以间接荧光方法(IFA)检测它们与Marc-145细胞的相互作用。IFA检测结果表明GST-GP5-1和GST-GP5-2融合蛋白均能够与Marc-145细胞结合,而且其结合位点在27aa～110aa区域。本实验为进一步研究GP5蛋白生物学功能提供试验依据。

Four GP5 gene fragments （79 bp to 207 bp, 133 bp to 330 bp, 229 bp to 492 bp, and 382 bp to 603 bp） from porcine reproductive and respiratory syndrome virus （PRRSV） （strain TA-12, NA type） were cloned into pGEX-6p-1 vector and expressed in E. coli BL21, respectively. The interaction of the GST-fusion proteins with Marc-145 cells were tested by indirect immunoflurouencent assay. The results showed that GST-GP5-1 and GST-GP5-2 could bound with Marc-145 cells, which indicated that the binding site of GP5 to Marc-145 cells was located at aa 27 to aa 101 of GP5.