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目的构建一种能预防AsiaⅠ型口蹄疫病毒(FMDV)的感染表位重组蛋白疫苗,并对其免疫学活性进行研究。方法以AsiaⅠ型FMDV VP1上的B细胞表位和T细胞表位氨基酸序列为抗原位点,设计了包含上述位点的、具有最佳空间构象的重组蛋白,命名为“CZ-1”。通过PCR及基因克隆等方法构建了其表达质粒,在大肠杆菌BL21(DE3)中诱导表达。通过镍亲和层析和G-25凝胶过滤层析纯化及复性,获得具有免疫学活性的重组蛋白CZ-1。免疫豚鼠获得含针对AsiaⅠ型FMDV中和抗体的血清,通过细胞中和试验及乳鼠中和试验,检测豚鼠血清中抗FMDV的保护性中和抗体滴度。结果该基因工程重组蛋白能够在动物体内诱导产生抗AsiaⅠ型FDMV具有保护性的中和性抗体。结论该基因工程重组蛋白有望开发成预防AsiaⅠ型FDMV感染的新型疫苗。
Objective To construct a recombinant epitope vaccine capable of preventing FMDV infection in Asia and to study its immunological activity. Methods Based on the amino acid sequences of B cell epitopes and T cell epitopes on Asia Ⅰ FMDV VP1, the recombinant protein containing the above site with the best spatial conformation was designed and named as “CZ-1” . The expression plasmid was constructed by PCR and gene cloning, and induced in E. coli BL21 (DE3). The recombinant protein CZ-1 with immunological activity was obtained by purification and refolding by nickel affinity chromatography and G-25 gel filtration chromatography. Immunization of guinea pigs Serum containing neutralizing antibodies to Asia type I FMDV was obtained and titers of protective neutralizing antibodies to FMDV in guinea pig serum were tested by cell neutralization and neonatal neutralization assays. Results The recombinant protein could induce neutralizing antibodies against Asia type I FDMV in animals. Conclusion The recombinant protein is expected to be developed into a new vaccine to prevent Asia Ⅰ FDMV infection.