Construction and Identification of a Recombinant Pseudorabies Virus Expressing ORF2 Gene of Porcine Circovirus Type 2

【Objective】In this paper,a recombinant virus expressing ORF2 gene of PCV2 using pseudorabies virus as vector.【Method】The ORF2 gene of porcine circovirus type 2 was inserted into the universal transfer vector deleting gG of pseudorabies virus to generate the recombinant transfer plasmid.The genomic DNA of PRV TK-/gG-/LacZ+ strain and pgGORF2 were co-infected into IBRS-2 cells.Then the recombinant virus TK-/gG-/ORF2+ was selected and its biological characteristic was measured.【Result】 The recombinant virus was selected by PCR with ORF2 gene and LacZ gene primers respectively and the PCV2 ORF2 gene had been inserted into the genome of TK-/gG-/LacZ+ strain identified by Southern blotting.The results of indirect immunofluorescene assay(IFA) and Western blotting indicated that the ORF2 gene was expressed successfully.Propagation of the recombinant virus in cells was corresponding to the parent strain.Animal experiments showed that the recombinant virus TK-/gG-/ORF2+ was safe to mice and could induce the specific antibodies against expressed ORF2 protein.【Conclusion】 The recombinant pseudorabies virus was constructed successfully,and the expressed ORF2 protein had immunogenicity.