Construction of a recombinant pseudorabies virus with BAC plasmid insertion in the TK gene.

In this study,a transfer vector pUCTK-EGFP-BACII was constructed by inserting the EGFP expression cassette and BAC plasmid pBeloBACII into two homologous sequences of TK gene of PRV Bartha K61 strain.The recombinant virus(rv-PRVBAC) was obtained by co-transfecting the transfer vector and PRV genome into Vero cell,followed by 11 passages of plaque purification based on EGFP expression.The rv-PRVBAC was shown to contain BAC plasmid and had similar growth profile in vitro as its parental virus Bartha K61,indicating the large fragment of BAC plasmid insertion in the TK gene didn't influence the replication of rv-PRVBAC in cell culture.The virus remained stable after 18 passages on Vero cell.The recombinant virus rv-PRVBAC reported here could serve as foundation for further study on bacterial artificial chromosome of PRV.