Preparation Of The Multiple Epitopes Vaccine Against Foot-and-Mouth Disease Virus Type A In Bovine

Foot-and-mouth disease is an acute, thermal and highly contagious diseases caused bythe foot-and-mouth disease virus in cloven-hoofed animals, which hinder the developmentof animal husbandry in the world. The disease has been defined as the OIE statutoryreported disease. To date, the mainly method to prevent and control the disease is toimmunize animals by vaccines. Inactivated vaccine plays a significant role in the controland elimination foot-and-mouth disease, but itself also has many defects, some developedcountries have already stopped the use of this vaccine. Developing a more safety, effectiveand new type vaccine become the most effective method to prevent and control the disease.The epitope-based vaccine as one of the most promising new vaccines showed goodexploitation and application prospect.To develop a new broad-spectrum multi-epitopes vaccine of type A foot-and-mouthvirus, The experiment is with our popular type A foot-and-mouth disease virus and ourcountry land neighbour popular representative strains (GenBank database) of VP1sequences in the past few years as the research object. With bioinformatics analysis to themajor antigen molecular, two main antigen epitope genes of structural protein waschosen in VP1,135aa~160aa and200aa~213aa as the main antigen Epitope regions,named as E1, E2. Single repeat of E1and E2alternating series and repeat contain of E1and E2were designed. After reconnection,3repeating series of DNA fragments werebuilt respectively with bovine IgG heavy chain.With the EcoRI/BamHI andBamHI/HindIIIsites, both of the two genes were cloned into the pET-30a (+) vector to form therecombinant plasmid pRE1IgG、pRE2IgG、p2REIgG, and then the recombinant plasmidswere transformed into E.coli BL21(DE3) competent cell. The SDS-PAGE and westernBlot results indicated that the fusion protein were highly expressed in inclusion bodies inE.coli after induced by IPTG, which can react with the positive serum against A typeFMDV.After quantitative, the above three types of recombinant protein RE1IgG, RE2IgG,2REIgG were mixed with ISA206in equal volume. guinea pigs were immuned in three groups. The three kinds of proteins of RE1IgG, RE2IgG and2REIgG were alsorespectively mixed with3D protein before mixed with equal volume. The recombinantprotein with3D protein as three other experimental groups, the control group wasdesigened and immune with qual volume of PBS. By indirect ELISA and virusneutralization test, results showed that compared with control group, all the experimentgroups can show ideal neutralization titer. However, there is no obvious improvement in3D plus group.