A pseudorabies virus (PRV) mutant with deletions in genes for glycoprotein I (gI) and glycoprotein E (gE) was constructed and evaluated as a candidate vaccine strain for its protective by virulent wildtype (wt) PRV Ea strain challenge in pigs. Transfer plasmid pIESE was constructed and co-transfected with PRV genome into IBRS-2 cells to generate a recombinant mutant PRV Ea gI-/gE- virus. The recombinant virus was confirmed by Southern blotting and indirect immunofluorescence assay (IFA). In animal experiments, the immunogenicity was tested by PRV-enzyme-linked immunosorbent assay (ELISA) and PRV neutralizing assay. PRV Ea gI-/gE- elicited significant humoral immune responses to wt PRV Ea, and PRV gI-/gE- immunization protected pigs against a lethal challenge by virulent PRV Ea strain. These results suggested that the recombinant PRV Ea gI-/gE- might be considered as a potential candidate vaccine against PRV.