Abstract: To study the location and function of EP0 protein in particles of pseudorabies virus, CDS region of EP0 gene was amplified by PCR, and then ligated with pMD18-T to construct a recombinant plasmid pMD-EP0.After identification by sequencing analysis, EP0 gene was extracted from this plasmid digested by EcoRⅠ and Hind Ⅲ, and cloned into pET-32a(＋)vector to construct a recombinant expression plasmid pET-EP0.This plamid was identified by PCR and sequencing analysis and then transformed into E. coli BL21(DE3).The target protein was detected by SDS-PAGE and Western blotting. The result showed that CDS region of EP0 gene, a fragment containing 1227 bp was amplified successfully and EP0 protein was expressed efficiently with the size of 75 ku and existed in inclusion body. The result of Western blotting showed that the expressed EP0 protein could be combined with positive serum of PRV.

Key words: pseudorabies virus; EP0 gene; cloning; expression