Construction Of The Eukaryotic Expression Plasmids With PPV VP2 Gene And PCV2 ORF2 Different Antigen Predominance Regionals And The Research On Immunogenicity

To study the immune efficacies,distributions and securities in animal bodies of recombinant eukaryotic expression plasmids with PPV VP2 gene and PCV2 ORF2 different antigen predominance regionals.VP2 gene was amplified from PPV SC-1 strain,and different antigen genes A,B,C(A:117～131aa,B:157～183aa,C:165～200aa) were amplified from PCV2 SC strain, respectively.The A,B,C genes and PPV VP2 gene were compounded in series and cloned into pCI-neo vector to construct the recombinant plasmids pCI-A-VP2,pCI-B-VP2,pCI-C-VP2.The recombinant plasmids were transfected into Vero cells to study the immune activities by immuno-fluorescent antibody technique. Healthy mice were inoculated with Porcine Parvovirus vaccine and Porcine Circovirus subunit vaccine, pCI-neo DNA and three recombinant plasmids for two times, the immune efficacies were detected by the MTT assay,FCM and ELISA. And the distributions,securities were inspected by PCR amplification.The results showed that the recombinant proteins were able to express nomally, fluorescence were observed in the cells transfected with pCI-A-VP2,pCI-B-VP2,pCI-C-VP2,and their immune efficacies were significantly higer than pCI-neo,and pCI-C-VP2 was of the higgest immune efficacies in the three recombinant plasmids. The PCR amplification results proved that the distributions were broad and the securities in animal bodies were good.These results provided reference datas for the bivalent vaccine research of PPV and PCV2.