Cloning And Expression Of EryA Gene Of Brucella Retrovirus Rev.1 Strain And Preparation Of Monoclonal Antibody

Brucellosis is a major zoonotic disease in the world.In recent years,brucellosis has shown an upward trend in China,which has caused great losses to the economy of our country.The gene structure,function and possibility of diagnosis of Rev.1 strain of Brucella mutans were analyzed in this study.The eryA gene was analyzed by bioinformatics analysis tool,expressed in E.coli,and used to prepare monoclonal antibody The results showed that the eryA gene was cloned and expressed successfully in this study.The coding protein could react with the standard positive serum specifically and show good reactionogenicity by Western-Blot.Bioinformatics analysis showed that the protein have no transmembrane structure,no signal peptide and the largest proportion of?-helix in the secondary structure.The antigenic epitope analysis showed that the protein contained more antigenic determinants.The hybridoma cells secreting IgG antibody were screened by fusion of mouse spleen cells after immunization with the purified protein.After expanded culture,mice were injected intraperitoneally to collect ascites and purify.The purified antibody was better,which detected by BCA and SDS-PAGE.This study provides a basic for the detection of brucellosis and the development of new products by studying eryA gene.