Prokaryotic Expression Of PRRSV ORF2 And ORF3 Gene In E.coli And Development Of Monoclonal Antibodies Against Protein GP2 And GP3

PRRS occurs in most major pig-producing areas throughout the word.The reproductive failure is characterized by abortions,stillbirths,and the birth of weak piflets that often die soon after birth of respiratory disease and secondary infection. In this study,after modifying of ORF2 and ORF3 gene of Porcine Reproductive And Respiratary Syndrom (PRRSV ) CH-1a strain ,we cloned it into plasmid pGEX6p-1 and expressed it in E.coli with IPTG inducing and named them rtGP2 and rtGP3 respectively. The molecular weights of rtGP2 is 40KU and rtGP3 is 42KU .They are active fusion protein and could be recognized by PRRSV positive serum in western blot.Then BALB/c mice were immunized with the purified recombinant fusion protein to develop monoclonal antibodies against these two fusion protein.The specificity of monoclonal antibodies were tested by ELISA assay.The results showed that one positive hybridoma cell lines stably secreting monoclonal antidody against GP2 were selected and named it 26D8.While obtain five positive hybridoma cell lines-31A2,31G11,32B12,32D4 and 32H7 stably secreting monoclonal antidody against GP3 .In indirect immunfluorescent assay, all of monoclonal antibodies reacted with PRRSV in weak fluorescent. The subtypes of all monoclonal antibodies are IgG1. Light chains of all monoclonal antibodies are kappa orescent.The result of our study will develop the bases of researching in GP2 and GP3, and provide a masure of analying about PRRSV antigene.