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

ORF2 gene of porcine circovirus type Ⅱ were cloned by PCR with the specific pri mers de-signed according to genome of PCV2 in GenBank.Following extraction and digestion,PCR productswere subsequentlyinsertedinto universal transfer vector pPI-2.EGFPto generate recombinant trans-fer plasmid pPI-2.EGFP-ORF2.The genomic DNAof PRV SA215 strain and pPI-2.EGFP-ORF2 were co-transfectedinto STcells withlipofectin,and recombinant virus SA215(C) was selectedby fluorescence and PCR with ORF2 gene pri mers respectively.The recombinant virus was analyzedwith Southern blotting and Western blotting.The results indicates that the insertion of EGFP andPCV2 ORFS gene does not affect proliferation of PRVSA215.Such a group of virus can be chosen asthe alternative attenuated vaccine strainfor PCV-2 and PRV.Protein can react with PCV2 positivesera.Results of virustiters detectionshowtheinsertion of ORF2 gene does not influence propagation ofrecombinant virus.