Abstract: A pseudorabies virus (PRV) transfer vector pLTK-HA containing a LacZ gene expression cassette and the HA gene of swine influenza virus (SIV) H3N2 subtype under the control of human cytomegalovirus (CMV) promoter was constructed and used to cotransfect with genomic DNA of strain Bartha-K61 into Vero cells using lipofectin transfection procedure. A recombinant PRV haboring SIV HA gene, designated as rPRV-HA, was generated after ten cycles of blue plague purification and PCR identification. The expression of the HA by rPRV-HA infected cells were characterized by Western blotting and indirect immunefluorescence assay (IFA). Compared to its parental virus Bartha-K61 strain, rPRV-HA showed no obvious difference in virus multiplication and cytopathogenic effects in Vero, PK-15, IBRS-2, chicken embryo fibroblast cells. The results of analyzing 30th passage rPRV-HA indicated that HA gene can be stably expressed by rPRV-HA.