口蹄疫病毒多抗原基因VP_1-VP_3-3C在大肠杆菌中的融合表达

Expression of FMDV VP_1-VP_3-3C Fusion Gene in E.coli

应用PCR扩增技术获得1564bp的片段,包含口蹄疫病毒VP1全长基因、VP3部分基因和编码3C裂解酶全长基因,连接到pMD-18T载体上,在获得重组质粒pMD-18T-P13C后,进行序列分析;并成功地构建了重组表达质粒PGEX-KG-P13C,将其转化大肠杆菌BL21(DE3)感受态细胞中,经IPTG诱导表达,收集菌体进行SDS-PAGE电泳。结果表明,P13C基因在大肠杆菌中获得成功表达,其表达产物为分子量83kDa的融合蛋白;能与猪抗FMDV血清发生反应,并为口蹄疫病的诊断及其基因工程疫苗的研究提供了依据。

A 1560bp DNA fragment, spanning whole VP1 gene, truncated VP3 and full length of 3C gene, was achieved using PCR and ligated to pMD-18T vector for sequence analysis. The recombinant expression plasmid was constructed by inserting the amplicon into PGEX-KG vector, followed by transformation of E.coli BL21 cells. The protein expression was induced by adding IPTG at final concentration of 1mM and was demonstrated by SDS-PAGE. The western blot showed the reactivity of the recombinant protein with swine anti-FMDV serum. These results provided the scientific implications in researches on the diagnosis and genetic engineering vaccine of FMDV.