Construction and Characterization Analysis of the Recombinant Virus SA215(C) Strain of Pseudorabies Virus-Porcine Circovirus Type 2

ORF2 gene of porcine circovirus type 2 were cloned by PCR with the specific primers designed according to genome of PCV2 in GenBank.Following extraction and digestion,PCR products were subsequently inserted into universal transfer vector pPI-2.EGFP to generate recombinant transfer plasmid pPI-2.EGFP-ORF2.The genomic DNA of PRV SA215 strain and pPI-2.EGFP-ORF2 were co-transfected into ST cells with lipofectin,and recombinant virus SA215(C) was selected by fluorescence and PCR with ORF2 gene primers respectively.The recombinant virus was analyzed with Southern blotting and Western blotting.The results indicated that ORF2 gene of PCV2 had been inserted into the genome of PRV SA215 strain and the expressed EGFP-ORF2 fusion protein could react with PCV2 positive sera.Result of virus titers detection showed that the insertion of ORF2 gene did not influence propagation of recombinant virus.