Abstract: A pair of primers was designed based on the DPV dUTPase gene sequence that our laboratory newly discovered (GenBank accession number DQ486149). Amplified PCR fragments were subcloned into the prokaryotic expression vector pET-32a (+). Then the recombinant plasmid pET32a-DU was transformed into E.coli BL21（DE3）strain and expressed under IPTG induction. SDS-PAGE analysis showed that the induced expressed protein was about 66.8 ku and accounted for 36.2% of total bacterial protein by gel scanning. The protein was purified and used to immunize rabbit for the dUTPase anti-serum production. Its ELISA antibody titer was up to 1∶409 600.Moreover, the subcellular localization detection was observed using immunofluorescence technique. The results showed that specific fluorescence appeared in cytoplasm as early as 4 hours post infection and a great deal of specific fluorescence concentrated in the cell nucleus by 12 hours. But after 24 hours, the fluorescence in nucleus was dispersed while that in cytoplasm increased. Forty-eight hours later, the fluorescence weakened significantly in both cytoplasm and nucleus. Above all, the results afforded significant data for the study on function of DPV dUTPase gene and the pathogeny of DPV.