Amplification and cloning of protein kinase gene of pseudorabies virus by PCR

Pseudorabies virus(PRV)was mumplicated on BHK-21 monolayer cells.After concentra-tion with centrifuge,viral DNA was isolated by SD-proteinase K digestion method.According to the sequences of protein kinase genes of PRV Ka and NIA-3 strains,the primers with 26 bpand 32 bp were designed.With pured PRV geneome DNA as template,the PK gene of PRV wasamplified successfully by PCR,and cloned into pUC19 vector.Restriction enzyme analysis showedthat the cIoned PK gene at Smal I,Xho I and Sal I site was the same as that of PRV NIA-3strain。This work laid foundation for constructing the PRV vaccine with deletion of PK gene.