摘要
流行性乙型脑炎病毒(JEV)可引起猪的繁殖障碍性疾病。本文利用基因重组技术,选取JEV prM/M、E、NS1蛋白的6个B细胞抗原表位,设计B细胞多表位基因,构建原核表达质粒,在大肠杆菌BL21中表达融合重组蛋白rMEP。纯化的rMEP免疫小鼠,通过抗体、中和抗体及亚型检测研究其免疫原性。结果表明,rMEP在上清中高效表达,Western blot结果证实rMEP具有较好的抗原性。在小鼠免疫模型中,rMEP免疫组小鼠血清中产生较高水平的针对rMEP的抗体和特异性中和抗体,且rMEP可诱导产生IgG1和IgG2a抗体,但以IgG1为主。这些结果表明,构建的JEV B细胞多表位蛋白能够刺激小鼠产生较强的体液免疫应答,为研究JEV的多表位疫苗提供新的思路和参考。
Japanese encephalitis virus （JEV） can cause swine reproductive disorder. In this study, a B cell muhiple-epitope gene, which comprised of six B cell epitopes from E, prM/M and NS1 protein of JEV, respectively, was designed using gene recombination technology. The muhiple-epitope gene was ligated into prokaryotic expression vector, and expressed in Escherichia coli BL21. Immunogenicity of the re- combinant muhi-epitope protein of JEV was investigated in mice. The result showed that recombinant fusion protein rMEP was effectively ex- pressed. Western blot analysis indicated that rMEP has good antigenicity to JEV. Mice immunized with rMEP produced the stronger humoral immune responses with high level of antibody titers and neutralizing antibody, compared to that of JEV vaccine control. ELISA resuh of anti- body isotypes indicated that IgG1 and IgG2a antibodies were induced and IgG1 was major subtype. The resuhs indicated that the recombinant muhi-epitope could induce strong humoral immune responses, suggesting that it might be an attractive candidate vaccine for preventing JEV infection.
出处
《畜牧与兽医》
北大核心
2014年第4期20-25,共6页
Animal Husbandry & Veterinary Medicine
基金
公益性行业(农业)科研专项项目(201203082)
江苏高校优势学科建设工程资助项目(PAPD)
关键词
流行性乙型脑炎
B细胞多表位基因
原核表达
免疫原性
Japanese encephalitis virus （JEV）
B cell muhi-epitope gene
prokaryotic expression
immunogenicity
