title: 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.
abstract: 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.
authors.affiliation: Departamento de Medicina Preventiva y Salud Pública y Microbiología, Facultad de Medicina, Universidad Autónoma de Madrid, Spain.
authors.forename: L
authors.initials: L
authors.lastname: Lerma
authors.affiliation: Departamento de Medicina Preventiva y Salud Pública y Microbiología, Facultad de Medicina, Universidad Autónoma de Madrid, Spain.
authors.forename: A L
authors.initials: AL
authors.lastname: Muñoz
authors.affiliation: Departamento de Medicina Preventiva y Salud Pública y Microbiología, Facultad de Medicina, Universidad Autónoma de Madrid, Spain.
authors.forename: S
authors.initials: S
authors.lastname: Wagner
authors.affiliation: Departamento de Medicina Preventiva y Salud Pública y Microbiología, Facultad de Medicina, Universidad Autónoma de Madrid, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Dinu
authors.affiliation: Departamento de Medicina Preventiva y Salud Pública y Microbiología, Facultad de Medicina, Universidad Autónoma de Madrid, Spain.
authors.forename: B
authors.initials: B
authors.lastname: Martín
authors.affiliation: Departamento de Medicina Preventiva y Salud Pública y Microbiología, Facultad de Medicina, Universidad Autónoma de Madrid, Spain. Electronic address: enrique.tabares@uam.es.
authors.forename: E
authors.initials: E
authors.lastname: Tabarés
mesh_terms: D000818:Animals; D002460:Cell Line; D022202:Chromosomes, Artificial, Bacterial; D004279:DNA, Viral / Q000235:genetics*; D020043:Genes, Essential; D005814:Genes, Viral; D006378:Helper Viruses / Q000235:genetics; D011558:Herpesvirus 1, Suid / Q000235:genetics* / Q000254:growth & development; D006801:Humans; D011995:Recombination, Genetic; D014774:Virulence
chemical_list: D004279:DNA, Viral
keywords: Construction recombinant pseudorabies viruses