Preparation And Potency Testing Of Subunit Vaccine Of The Staphylococcus Aureus From Matitis In Dairy Cows

The bovine mastitis, as one of the most common and multiple diseases, not only cause seriouseconomic loss in breeding industry, but also does endanger human health. Staphylococcus aureus is one ofthe most popular pathogeneses. In view of drug residue and drug resistance of aureus, the vaccine becomesthe most effective methods for the prevention of bovine mastitis.The adhesion is a protein located in bacterial surface, which could be identified and combine withtissue and cells. The capsular polysaccharide is an important surface structure for survival of aureus in hostand environment, which will become a T cell-dependent antigen when a carrier protein coupled with.Almost all the staphylococcus aureus has ibronectin-binding protein and clumping factor A. The region ofA is the most functional area for clumping factor A. The type-5capsular polysaccharide is the mainserotype in clinic. In this study, these three virulence factors were used for vaccine research.Objective: Preparation of Subunit Vaccine of the Staphylococcus Aureus from Matitis in Dairy CowsMethod:ClfA A and FnBPA genes were amplified from Whole Genome of SHZ1strain usingPolymerase Chain Reaction and the fusion protein was obtained by tandem expression in vector pET-32a.CP5was separated by the high Pressure Technology and purified by enzyme and DEAE Sephacel ionexchange chromatography. The polysaccharides content was detected by the phenol-sulfuric acid method.And the content of protein and nucleic acid were detected by instrument. The mixture of CP5and fusionprotein was joined by EDAC and successful by confirmation of the ultraviolet scanning spectrometer. Theconjugation ratio of CP5and protein was detected by the phenol-sulfuric acid method.Results:The result of sodium dodecyl sulfate polyacrylamide gel electrophoresis showed that themolecular weight of fusion protein was about100kDa. The expression level of fusion protein wasdetected by Western-blot technology.The polysaccharides content was detected by the phenol-sulfuricacid method. And the content of protein and nucleic acid were detected by instrument. There was noanti-idiotype antibody in the immunological test of CP5.The immunologic tests of mixture showed betterimmunogenicity and protective immunity against staphylococcus aureus induced in mice. It supposed tobe a new subunit vaccine of staphylococcus aureus.Conclusion: Better immunogenicity and protective immunity against staphylococcus aureus showedthat the mixture of fusion protein and CP5maybe be a new subunit vaccine of staphylococcus aureus.