The Study On B, T Cell Antigen Epitope EpiW-GST Polypeptide Vaccine Of Avian Infectious Bronchitis Virus

IB is one of the great infectious diseases which harmed the poultry industry. At present the domestic and oversea use the ambly-poison vaccine and deactivation vaccine in common, but the mutation of IBV genomic that has much serological type and was difficult to prevent. It is necessary to request effective vaccine to prevent and cure IB. Polypeptide vaccine were conservation and security, has already become a new hot spot on vaccine development in the future.The study at the based on the EpiW which designed by our laboratory, it was a gene own IBV B, T cell antigen epitope. At first, predictive analysis and probation the primary structure, hydrophilicity, hydrophobicity, antigenicity, enzyme inside sites, spatital structure of the EpiW, to ensure the exact in abstracto.Secondly, Then EpiW were inserted into the multiple cloning sites (MCS) of the prokaryotic expression vector pGEX-4T-l. Restriction enzyme analysis and identification of PCR showed that were constructed successfully, then transforming it into competent cell BL21, screening masc. clone, induce express and rep the production to analyze. The result manifest that the EpiW recombination gene was be successed express. Used the GST purification the recombination protein, which as the coating antigen, detected the IBV stand masc. Serum. Initial study indicate that recombination protein could create immune reaction with masc. Serum, to advance adopt the western blot, show that they could reaction specific reaction, testimonial the EpiW owned the biology-activity.In order to evaluated the immunes effect of EpiW-GST polypeptide vaccine, recombination protein EpiW-GST was purified and did animal immunity tes to match with FA, inactivation vaccine, GST, PBS was to be control group, detect the cell immunity that deploy the FACS which det the percentage of CD4,CD8+T lympy cell in peripheral blood. IBV specific antibody was detect by use ELISA, lastly, done the eliminating toxic material experiment.The results indicated that the titers of peculiarity antibody of the were higher than other control experiment groups (P<0.05). However, the percentage of CD₄+ and CD₈+ T lymphocyte of the EpiW was higher than the control experiment groups r (P<0.05). The protection rate of EpiW group was 80%. Concluded that. it offer the new way of the study on IBV vaccine that the E coli biosynthesis recombinants IBV polypeptide vaccine. This study provides some basic material and scientific gist for the further development of prevent and control against IBV.