摘要
本试验通过自行设计两段引物,利用重叠延伸PCR方法,将伪狂犬病病毒(PRV)gB基因两段优势抗原表位序列串联,克隆至pMD18-T载体,测序正确后双酶切连接至pET-32a(+)载体,构建重组质粒;重组质粒转化入大肠杆菌BL21(DE3)受体菌,经IPTG诱导后,通过SDS-PAGE和Western blotting检测融合蛋白表达情况。经检测,诱导后的重组蛋白获得表达,重组蛋白大小约为54ku,其中串联蛋白大小约为34ku。该重组蛋白可与伪狂犬病病毒gB蛋白单克隆抗体发生特异性反应,表明重组蛋白的抗原性良好。
The two dominant epitope sequences of pseudorabies virus （PRV） gB gene were joined together by overlap-extension PCR method with two pairs of primers. The products were inserted into pMD18-T vector. After properly sequenced, the multi-epitope sequence was linked with pET-32a（＋） vector,and then transformed into BL21（DE3）. By being induced with IPTG,the targeted protein was detected by SDS-PAGE and Western blotting. The results showed the fusion protein was successfully expressed with the size of 54 ku including 34 ku tandem protein and could be combined with monoclonal antibody against gB which showed that the expressed protein had good antigenicity.
出处
《中国畜牧兽医》
CAS
北大核心
2015年第4期810-815,共6页
China Animal Husbandry & Veterinary Medicine
基金
国家自然科学基金(31001074)
