Abstract:The Pseudorabies virus (PRV) Fa strain gC gene was subcloned into adenovirus shutter vector pDC316 through double enzymatic digestion. After cotransfection of the shutter vector pDC316-gC and adenovirus DNA helper plasmid pBHGloxΔE1,3Cre into 293 cells, the recombinant adenovirus expressing gC protein was preliminarily obtained through homologous recombination with observation of remarkably cytopathic effect. Afterwards, high titer of recombinant adenovirus designated as Adv-gC were prepared after one round of PCR identification and virus plaque purification. Glycoprotein gC gene expression was confirmed in Marc145 cells by immunofluorescent staining assay. Intramuscular immunization of Adv-gC could induce PRV specific humoral and cellular response with assays of anti-gC protein IgG antibodies、neutralizing antibodies and delayed-type hyper-sensitivity(DTH) in mice. In the challenge experiment, the group immunized by Adv-gC presented 100% protection efficiency as contrast to 0% obtained in the negative control group. Totally, the present result will provide a good foundation for developing a novel type of PRV live vector vaccine.