鲤春病毒血症病毒糖蛋白的高效表达和纯化

本研究从鲤春病毒血症病毒（Spring viremia of carp virus，SVCV）SVCV-741毒株克隆鲤春病毒血症病毒糖蛋白基因（sue—g），将svc-g亚克隆到原核表达载体pET-28a，转化大肠杆菌E．coli BL-21（DE3）后，用IPTG诱导培养，获得菌体总蛋白。用SVCV毒株免疫山羊所得到的抗血清作为一抗进行免疫印迹实验，结果在硝酸纤维素滤膜上检测到50～71 kDa之间的特异性免疫条带，与SVCV糖蛋白预测分子量一致，研究证明了工程茵表达获得的重组蛋白具有与SVCV毒株相同的免疫原性，也证明了该蛋白的糖基化对其免疫表位是非必需的。本研究进一步通过发酵基因工程菌和蛋白质纯化过程，获得大量的鲤春病毒血症病毒糖蛋白，为后期免疫动物获得抗血清储备了原料，也为SVCV的免疫学检测方法的建立奠定了物质基础。

HIGH-LEVEL EXPRESSION AND PURIFICATION OF RECOMBINANT GLYCOPROTEIN OF SPRING VIREMIA OF CARP VIRUS (SVCV)

Glyeoprotein gene （sve-g） of Spring viremia of carp virus （SVCV） was cloned from isolate SVCV-741. The svc-g gene was subcloned into plasmid pET-28 and the resultant plasmid was transformed into E. coli BL-21 （DE3）. The total protein was obtained from E. coli cultures induced by IPTG. Westeru blot using goat anti-SVCV serum revealed a specific band between 50 kDa and 71 kDa. This proved that target protein has the same immunogenicity as SVCV strain. It is speculated that glycosylation is not necessary for the epitopes of glycoprotein. A large quantity of recombinant glycoprotein can be produced by fermentation of recombinant bacteria.