Construction And Immunogenicity Of Recombinated Goat Pox Virus Expressing Omp31 And Omp2b Gene Of Brucella

Brucellosis is a zoonosis ,which is caused by gram-negative Brucella that parasitizes in cell. The symptoms of Brucellosis includes fetal death, abortion and sterility of livestock. This disease not only affects the development of animal husbandry but also causese enormous economy losses and serious public health problem. For the expection to obtain recombinated Goat Pox virus which would have strong immunity to Brucellosis, in this study, recombinated Goat Pox virus which contained Brucella antigen omp31 gene and omp2b gene was constructed on the base of our country's Goat Pox vaccine which was used widely. This study would provide an experiment and theory basis for the prevention of Brucellosis and Goat Pox.According to the published sequences of sheep species Brucella omp31 gene and omp2b gene on GenBank, two pairs of primers were designed. The omp31 gene and omp2b gene were amplified and ligated to the T vector, which were identified by enzyme cutting and sequencing. The right sequences of omp31 gene and omp2b gene were ligated with GTPV vector pGM-TK-P11-GPT1 for constructing transfer vector. The transfer vectors and GTPV vaccine were co-transfected into sheep testicular cells and the recombinant virus were identified by screening, After analyzing the OMP31 and OMP2b protein expressed by recombinant virus by Western blot, the in vivo IgG level of mice immuned with rGTPV were detected by indirect ELISA.In this experiment,two recombinated Goat Pox virus expressing Bucella OMP31 and OMP2b protein stably were constructed.. Western blot analysis showed that the OMP31 and OMP2b protein expressed by recombinated virus had immune reactivity. After the mice were immuned by rGPTV , the OMP31 and OMP2b protein expressed by recombinated virus can produce corresponding antibody. The study provided a technology platform for further study on GPV live vector vaccine and related foundation research.