Construction of Recombinant Adenovirus Co-expressing PCV-2 ORF2 and PRRSV GP5 Genes and Immunoassay in Mice

The recombinant shuttle plasmid was constructed successfully after the PCV-2 ORF2 and PRRSV GP5 genes were linked by IRES gene.After being digested with PmeⅠ,the linearized recombinant plasmid was transformed into Escherichia coli BJ5183 competent cells.The homologous recombination between the linearized plasmid and the supercoiled adenoviral vector pAdEasy-1 occurred.The recombinant adenovirus plasmid was digested with PacⅠ and transfected into 293A cells to pack into viral particles.The recombinant adenovirus was acquired.The constructed recombinant adenovirus containing ORF2 and GP5 genes was identified by PCR.The Western blot results showed that ORF2 gene of PCV-2 and the GP5 gene of PRRSV were expressed in 293A cells successfully.The recombinant adenovirus was passaged 30 times in 293A cells.The titer was stably 10-9.5/mL.Furthermore,the recombinant adenovirus could induce PCV-2 and PRRSV specific humoral immunity in mice.Therefore,the recombinant adenovirus may be a candidate for PCV-2 and PRRSV gene engineering vaccines.