Construction of recombinant pseudorabies viruses by using PRV BACs deficient in IE180 or pac sequences: Application of vBAC90D recombinant virus to production of PRV amplicons

•Construction of PBAC80 deficient in pac sequences and inducuble IE180 expression.•A simple strategy to generate of PRV recombinant viruses in mammalian cells is described.•PRV-BT80GF virus, that expresses EGFP protein, was obtained by using PBAC80.•vBAC90D virus, deficient in IE180, was obtained by using PBAC90.•vBAC180D is useful as a helper in the production of PRV amplicons without cell toxicity.

We describe a simple and efficient method to obtain recombinant pseudorabies virus (PRV) in mammalian cells by using the PRV BACs, PBAC80 deficient in pac sequences and PBAC90 deficient in the IE180 gene. These essential viral sequences were used as targets to obtain viable recombinant viruses. PBAC80 was constructed, confirmed to encode a copy of the IE180 gene regulated by the inducible Ptet promoter, and used to obtain recombinant attenuated PRV viruses that express the EGFP protein (PRV-BT80GF virus). PBAC90 was used to obtain the vBAC90D virus, deficient in IE180 and free of replication-competent revertants, and which can be used as a helper in the production of PRV amplicons.