Cloning of the gD and gE of pseudorabies virus and construction of a universal transfer vector

The gD and gE gene of the pseudorabies virus Min-A strain were obtained by polymerase chain reaction(PCR),and then cloned into the pGEM-T Easy vector.The gD and gE gene is subcloned into pUC18,resulting in pUgDgE.The fragment from pcDNA3.1-including hCMV promoter/enhancer,MCS and neomycin resistance gene was inserted into the BamHI and BstEII sites of pUgDgE,resulting in the universal transfer vector pgD-M-gE.The universal transfer vector pgD-M-gE deleted the gI gene and 363 bp in the 5' end of the gE ORF of PRV.There were 11 restrication sites for insertion of the foreign gene.The upstream and downstream flanking sequences were up to 1.25 kb and 1.42 kb.It will be useful for developing the recombinant PRV expressing foreign gene(s).