Abstract: In order to construct the recombinant Pseudorabies virus strain expressing the out membrane CD2 v protein（encoded by EP402 R gene） of African swine fever virus（ASFV） and study its biological characteristics, this experiment used homologous recombination and ccdB counter-selection technique to replace TK gene of pBeloBAC11-Bartha-K61 vector with EP402 R gene of PUC57-CMV-Flag-EP402 R-BGH plasmid（the EP402 R belonged to Georgia 2007/1 strain, GenBank accession number FR682468） and construct pBeloBAC11-Bartha-K61-（TK）CMV-Flag-EP402 R-BGH recombinant plasmid. Then, the recombinant plasmid was transfected into Vero cells for virus rescue, and the recombinant Pseudorabies virus strain rBartha-K61-EP402 R was obtained through cytopathic observation, PCR and Western-blot identification; its stability and proliferation characteristics were studied. The results showed that the EP402 R gene and Flag tag protein of the constructed recombinant Pseudorabies virus strain after 20 consecutive passages could be still detected. The proliferation curve of the recombinant virus was consistent with that of the parent strain, and the peak value of the virus titer could reach 1×10^5.6TCID₅₀/mL. The results suggested that the experiment successfully constructed a recombinant Pseudorabies virus strain expressing the outer membrane protein CD2 v of ASFV,and the strain had good genetic stability and proliferation performance.