Construction And Immunogenicity Study Of Fusion Eukaryotic Expression Vector Of Porcine IFN-a Gene, PPV VP2 And PCV2 ORF2 Gene

Construction and immunogenicity study of fusion eukaryotic expression vector of porcine IFN-a gene,PPV VP2 and PCV2 ORF2 gene Tan Rong(Preventive Veterinary Medcine) Directed by Prof. Wang Yin(Sichuan Agriculture University, Ya'an, Sichuan, P.R.China,625014)Interfron was a kind of cytokine with Broad-spectrum antiviral and immunoregulation activity, generally, it secretes by host cell when invaded by interferon inducer. Previous research shown that porcine interferon a can be used as immunopotentiator of vaccine and medicine of porcine virus disease. With the increasing development of genetic engineering technology, more and more studies focus on the application of porcine interferon a as a cytokine adjuvant, and develop new DNA vaccine with recombinant IFN-a gene.Utilizing technique of PCR and sequencing, the nucleotide and deduced amino acid sequence of the porcine IFN-a gene were obtained in this study. Three nucleotide mutations in CDS region were detected, two of them are missense mutations.Fusion expression expression vector were constructed using plasmid pMD-IFN-a and pCI-VP2-ORF2.High level expression of recombinant plasmid were detected by indirect immunofluorescence and ELISA in transfected PK15 cell. Then recombinant plasmid pCI-IFN-a-ORF2-VP2 were intramuscular injected in BALA/c mice as a DNA vaccine. Well immune response were induced, pCI-IFN-a-ORF2-VP2 result in higher PPV/PCV2 antibody titer and cellular immunity level than pCI-ORF2-VP2 and other vaccine.The experiment was conducted to study the immunomodulatory effects of insertion of cytokines IFN-a as adjuvant, the recombinant plasmid PCI-IFN-a-ORF2-VP2 eukaryotic expression vector DNA vaccine and its immunogenicity and safety of research, genetic engineering for the PPV and PCV2 vaccine polyvalent lay the foundation for further study.