Abstract: Through digestion, filllinking method to transform PUSK vector to eliminate the vector of BamHⅠ and HindⅢ restriction sites. According pAcGFP1C1 gene sequencing to designe primers, in the upstream and downstream primers 5′end introduced NotⅠ restriction sites, will be amplified by PCR fluorescent protein gene AcGFP(including CMV, MCS, SV40 Poly (A)) cloned into the vector NotⅠ restriction sites of the PUSK. Use of lipidmediated method, vector plasmid PUSG and PRV genomic DNA transfection of PK15 cells. Green fluorescent will be observed in 48 hours under fluorescence microscopy. After three consecutive passages have been purified virus. The recombinant virus will be providing a convenient for the next two or trivalent vaccine selection.

Key words: pseudorabies virus; PUSK; gG gene; pAcGFP1C1; transfer vector; expression