牛AsiaⅠ型口蹄疫病毒重组蛋白疫苗的构建及功能研究

Construction and functional study of recombinant vaccine for bovine AsiaⅠfoot-and-mouth disease virus

目的为了取代有安全隐患的牛AsiaⅠ型口蹄疫病毒灭活疫苗,我们构建了基因工程重组蛋白疫苗。方法应用PCR方法合成含有表位的基因片段,经过克隆连接获得重组基因,在大肠杆菌中表达后经镍亲和层析纯化重组蛋白。重组蛋白免疫豚鼠后,分别经ELISA和乳鼠中和实验检测血清中抗FMDV抗体水平。结果构建了牛AsiaⅠ型口蹄疫病毒重组蛋白疫苗的结构基因,并成功表达和纯化了该重组蛋白。功能实验表明,该蛋白在豚鼠体内诱生了高水平的抗牛AsiaⅠ型口蹄疫病毒的中和性抗体。结论该重组蛋白为制备牛AsiaⅠ型口蹄疫病毒新型疫苗提供了有价值的线索。

In this study,we tried to develop a recombinant protein vaccine for preventing bovine Asia I foot-and-mouth disease virus infection.One B cell epitope and two T cell epitopes from FMDV Vp1 protein were chosen to construct genes encoding recombinant proteins RA2.And then RA2 gene（consisting BT2-BT2-BT1-BT2-BT2） was constructed by PCR and DNA cloning,which was subcloned into pET28a and highly expressed in BL21（DE3）.RA2 protein was purified by nickel chromatography and immunized in genuine pigs. ELISA and suckling mice protection method found ligh level anti-FMDV antibody in sera of the immunized genuine pigs.The results imply that the recombinant bovine Asia I foot-and-mouth disease virus vaccine RA2 may be developed as a safe and effective novel vaccine.