猪繁殖与呼吸综合症GP5蛋白的纯化与免疫活性分析

1材料与方法1．1表达菌及试剂猪繁殖与呼吸综合征病毒（PRRSV）6P-1-GP5表达质粒由该实验室保存。Novagen蛋白纯化试剂盒、硝酸纤维素膜（NC膜）均购自华美公司；弗氏完全佐剂、弗氏不完全佐剂购自Sigma公司；辣根酶标记兔抗猪IgG购自北京中杉金桥生物技术有限公司；其他常规试剂均由国内生产。试验用豚鼠由中国农业科学院兰州兽医研究所实验动物厂提供，体重400-500g，共8只。

Purification and Immunocompetence Analysis of GP5 Protein in Porcine Reproductive and Respiratory Syndrome

Objective] The purification and immunocompetence of GPS protein in porcine reproductive and respiratory syndrome (PRRS) were analyzed in this study, which provided basis for establishing the corresponding serological method. Method] The recombinant expression plasmid pGEX-6P-5 was transformed into BL21 and expressed after being induced with IPTG. The solubility analysis of expression products was carried out, and then the recombinant protein was purified for SDS-PAGE identification and Western-blot analysis. Finally, the recombinant antigen was used in the immune experiment of guinea pigs. Result] The target protein content accounted for 30% of the total cells protein content according to the chromatography scanning, and the purity of target protein after purification reached 80%. The purified protein was analyzed by Western-blot and immune experiment of guinea pigs, and the results showed that the expressed protein had good reactionogenicity and immunogenicity. Conclusion] This study provides materials for further studies on the function between PRRSV ORF5 gene and its editing protein, which also lays a foundation for porcine reproductive and respiratory syndrome virus genetic engineering products.