论文部分内容阅读
为获得可用于诊断的猪瘟病毒(Classical swine fever virus,CSFV)E2蛋白重组抗原,对CSFV-E2的多个B细胞抗原表位进行重构和表达。将人工合成的E2蛋白多抗原表位基因与p ET-28a(+)载体连接后,转化至宿主菌BL21(DE3),IPTG诱导表达,SDS-PAGE电泳分析蛋白可溶性,His亲和层析柱纯化蛋白并进行抗原性检测。重构后的猪瘟病毒E2基因多抗原表位基因大小约500 bp,PCR、双酶切鉴定结果显示与预期相符;重组蛋白为可溶性表达,大小约23 k Da;Western blot结果显示,重组蛋白可与猪瘟阳性血清发生特异性结合,具有良好的免疫反应性,可作为诊断抗原用于猪瘟病毒感染动物的血清抗体检测。
To obtain recombinant E2 antigen of classical swine fever virus (CSFV) that can be used for diagnosis, multiple B cell epitopes of CSFV-E2 were reconstructed and expressed. The synthetic E2 antigen epitope gene was ligated with p ET-28a (+) vector and transformed into host strain BL21 (DE3). The recombinant protein was induced by IPTG and analyzed by SDS-PAGE electrophoresis. His affinity chromatography column Purify the protein and perform antigenic testing. The reconstructed E2 gene of classical swine fever virus was about 500 bp in length. The PCR and restriction enzyme digestion showed that the gene was consistent with the expectation. The recombinant protein was soluble and expressed about 23 kDa. Western blot showed that the recombinant protein Can be combined with classical swine fever serum, has good immune reactivity, can be used as a diagnostic antigen in swine fever virus infected animals serum antibody detection.