PCV2 ORF2基因在大肠杆菌中的表达及与PCV2灭活抗原免疫效果的比较

利用基因克隆方法,将PCV2的ORF2基因PCR扩增并酶切后连入pET-32a载体,构建重组质粒。重组质粒经PCR和酶切鉴定并测序后,转化大肠杆菌BL21进行IPTG诱导表达。表达产物经SDS-PAGE和Western Blotting分析鉴定后,与PCV2灭活抗原分别与弗氏不完全佐剂乳化,并分别免疫仔猪以比较免疫效力。结果为PCV2的Cap蛋白得到正确表达,免疫保护性试验结果显示,相对于灭活抗原制备的疫苗,表达的Cap蛋白制备的疫苗具有更好的免疫原性和保护效力。

Expression of Gene ORF2 of PCV2 in E. coli and Comparison of Immune Effect between It and Inactivated Porcine Circovirus 2 Antigen

In this study,the gene ORF2 of PCV2 was amplified by PCR and cloned into the pET-32a plasmid through enzyme digestion for the construction of the recombinant plasmid.After identification and sequencing,the recombinant plasmid was transformed into E.coli BL21 competent cells to induce the expression of Cap protein of PCV2.After confirmation by SDS-PAGE and Western Blotting assays,the expressed products and inactivated PCV2 viruses were emulsified with IFA to make vaccines.The immune efficacies of the two vaccines were compared in pigs.The results showed that the Cap protein was expressed correctly.As compared with the vaccine made by inactivated PCV2 viruses,the vaccine made by Cap protein showed better immunogenicity and protective efficacy.