The Study Of Antigen Immunological Properties And Screening Of Advantaged Epitope Gene Of Brucella

As a zoonotic infectious disease, brucellosis has been a serious threat to the stabilityand development of the animal husbandry and economy. The disease is classified as twoClass infectious disease in China, that caused by Brucella infection. Brucella infectedanimals suffer even worse effects such as abortion, infertility, and tissue necrosis. For thepast years, the infection and morbidity rates stay high in some areas in our country and allover the world. Currently, few thorough cures have been reported around the world, andthe most effective prevention is vaccination, it is particularly important for research andprevention and control work about Brucella and brucellosis.The outer membrane protein (Omp) of brucella is surface exposured to bacteria, it hasbeen regarded as a research which not only can react with IgG antibody, but also can causea protective immune response, the cytoplasm binding protein is main composition ofantigen determinant for Brucella, which related to the pathogenicity of Brucella. Thepresent study focused on a gene about cytoplasm binding protein P39and the genes(omp10, omp19, omp22, omp25and omp28) about outer membrane protein from theBrucella, cloning the genes by PCR amplified using primers with genomic DNA ofattenuated live vaccine S2of B. suis as template. The target gene were inserted into cloningvectors resulted in recombinant plasmids including pET-28a-omp10, pET-28a-omp19,pGEX-4T-1-omp22, pET-28a-omp25, pET-28a-omp10and pGEX-4T-1-P39, they can beexpressed proteins in E. coli strain TL129or BL21under an IPTG inducible promotercontrol and the expressed proteins were detected by SDS-PAGE. The proteins expressed inE.coli were purified by GST column, the purified proteins were used to inoculatelong-eared rabbits by injection. The antiserums were collected and the immunogenicitieswere analysed by Western blot. Results showed that the recombinant proteins includedOmp10, Omp19, Omp22, Omp25, Omp28and P39molecular weight for38.5kDa,17.6kDa,47kDa,53.5kDa,52.5kDa and39kDa, in line with expettations. The antiserumsthat were prepared rely on the recombinant proteins Omp22and P39against the twoproteins had intense immune response. the recombinant proteins Omp22and P39hadstrong immunogenicity. This study provided theoretical and experimental basis for the further development ofgene vaccine and recombinant subunit vaccine.