Cloning and expression of the Staphylococcus hyicus ExhB gene in E.coli and its function analysis

Cloning,expression Staphylococcus hyicus ExhB gene which can cause exudative epidermitis in the piglet and identification the immunity of the protein encoded by ExhB gene,and then the analysis of the biological function of the expression products.The Staphylococcus hyicus genomic DNA was PCR amplified by the primer1 and primer 2.We can get the ExhB gene in this way.After this gene was identified by clone and sequence analysis,it was cloned in the plasmid vector pET32a.The recombinant bacteria was transferred into BL21 in order to realize its expression.The protein was purified by affinity chromatography.We also inject the recombinant protein into the nude mice and identify its biological activity.Amplified the S.hyicus ExhB gene and purified the recombinant protein.SDS-PAGE showed the molecular weight of recombinant protein was 45 000.Most of the protein was existed in including body.The ELISA and agar pervasion experiment indicate that the recombinant protein could combination with the polyclonal mice antibodies immunized by the S.hyicus.The recombinant protein can cause histological change in nude mice.Successfully construct the recombinant protein.The protein could react with the polyclonal serum.The nude mice are susceptible to the protein too.This experiment can be the basis for the more deep research such as the study of the pathogenesis of S.hyicus and the exactly treatment and prevention method.