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为构建重组牛O型口蹄疫病毒(FMDV)VP1表位疫苗,并对其免疫学活性进行研究。利用计算机模拟构象的方法筛选出牛O型FMDVVP1表位六聚体重组蛋白的最佳表位组合形式,通过PCR及基因克隆等方法构建含编码该重组蛋白基因的质粒,大肠杆菌BL21(DE3)中诱导表达,镍亲和层析法纯化。通过间接ELISA方法初步检测该蛋白免疫豚鼠血清中的抗FMDV抗体水平;再用活病毒FMDV进行细胞中和实验和乳鼠保护实验以检测豚鼠血清中具有保护性的抗FMDV中和性抗体的滴度。结果:构建了一种编码牛O型口蹄疫病毒VP1表位六聚体重组蛋白的质粒,经诱导表达并纯化后得到重组蛋白,命名为“MIP10”。ELISA检测结果显示,MIP10蛋白免疫的豚鼠血清中含有高水平的抗牛O型FMDV抗体。细胞中和实验和乳鼠保护实验结果显示,MIP10蛋白免疫的豚鼠血清中含有针对牛O型FMDV的保护性抗体。重组牛O型口蹄疫病毒VP1表位疫苗MIP10能够在动物体内诱导产生具有保护性的抗牛O型FMDV中和性抗体,有望开发成预防牛O型FMDV感染的新型疫苗。
To construct recombinant VP1 epitope vaccine of FMDV and to study its immunological activity. The optimum epitope combination of hexamer-FMDV VP1 epitope was screened by computer simulation conformation, and the plasmid containing the recombinant protein gene was constructed by PCR and gene cloning. E.coli BL21 (DE3) Inducible expression, purified by nickel affinity chromatography. The level of anti-FMDV antibody in the serum of guinea pigs immunized with the protein was detected by indirect ELISA. The cells were neutralized with live virus FMDV and the suckling rat protection test was used to detect the protective anti-FMDV neutralizing antibody in guinea pig serum degree. Results: A recombinant plasmid encoding the VP1 epitope of bovine O-type foot-and-mouth disease virus (VP1) was constructed and expressed as a recombinant protein. The recombinant protein was named “MIP10”. The results of ELISA showed that MIP10 protein immunized guinea pig serum contained high level of anti-bovine O FMDV antibody. Cell neutralization and suckling protection experiments showed that guinea pig serum immunized with MIP10 protein contained a protective antibody against bovine O FMDV. Recombinant bovine O-type foot-and-mouth disease virus VP1 epitope vaccine MIP10 is capable of inducing the production of a protective neutralizing anti-bovine FMDV antibody in animals and is expected to be developed into a new vaccine against bovine O FMDV infection.