The envelope glycoprotein E is a major glycoprotein of pseudorabies virus, which exerts important effect in pseudorabies eradication campaign. The gE gene fragment deleted signal peptide of PRV Fa strain was inserted into Pichia pastoris expression vector pPICZαA, the resulted recombinant expression vector transformed SMD1168 competent cells and obtained engineering Pichia pastoris strain SMD1168/pPICZαA-FL. After high concentration Zeocin^TM screening, phenotype identification, inductive expression, SDS-PAGE and Western blot analysis of culture supernatant, an engineering Pichia pastoris strain SMD1168/pPICZαA-FL-7 in which gE gene fragment deleted signal peptide was expressed in high levels was obtained. SDS-PAGE and Western blot indicated that expression product of gE fragment deleted signal peptide in culture supernatant of SMD1168/pPICZαA-FL-7 was about 80 ku, a little larger than expected. Gel scanning and Bradford protein analysis results showed that expression product reached 11.7 g/L, or 13.49% of total culture supernatant protein in SMD1168/ pPICZαA-FL-7.