title: Characterization of interspecific recombinants generated from closely related bovine herpesviruses 1 and 5 through multiple PCR sequencing assays.
abstract: Bovine herpesviruses 1 (BoHV-1) and 5 (BoHV-5) are closely related alphaherpesviruses infecting cattle. In countries where both viruses circulate, co-infection of cattle is likely. It was shown that recombination occurs at a high frequency in cattle infected dually with two BoHV-1 mutants. In addition, interspecific recombinants are generated in cell culture co-infected with BoHV-1 and BoHV-5. Even if the process of interspecific recombination appears inefficient relative to intraspecific recombination, BoHV-1 and BoHV-5 may give rise to interspecific recombinants in co-infected cattle. Since molecular tools for differentiating BoHV-1 from BoHV-5 are limited and do not allow to localize recombination events between these closely related virus species, 13 PCR sequencing assays were developed to discriminate between BoHV-1 and BoHV-5 at regular intervals throughout the entire respective viral DNA genomes. These assays were used to determine the genetic background of two interspecific BoHV-1/-5 recombinants generated previously. The two crossover points where recombination events occurred between the parental strains were determined. This study provides a detailed analysis of two interspecific recombinant viruses generated in vitro from closely related alphaherpesviruses infecting the same natural host. It demonstrates that recombination can occur within very short fragments of sequence homology. This finding raises questions about the mechanisms involved in the strands exchange and resolution step of the homologous recombination used by herpesviruses. This method will allow monitoring generation of recombinants between closely related herpesvirus species both in vitro and in vivo.
authors.affiliation: Virology Institute, Veterinary and Agricultural Science Research Centre, National Institute of Agricultural Technology, Castelar, Buenos Aires, Argentina.
authors.forename: Maria Paula
authors.initials: MP
authors.lastname: Del Médico Zajac
authors.forename: Sonia Alejandra
authors.initials: SA
authors.lastname: Romera
authors.forename: Maria Fatima
authors.initials: MF
authors.lastname: Ladelfa
authors.forename: Fiorella
authors.initials: F
authors.lastname: Kotsias
authors.forename: Julien
authors.initials: J
authors.lastname: Thiry
authors.forename: Dominique
authors.initials: D
authors.lastname: Ziant
authors.forename: François
authors.initials: F
authors.lastname: Meurens
authors.forename: Günther M
authors.initials: GM
authors.lastname: Keil
authors.forename: Etienne
authors.initials: E
authors.lastname: Thiry
authors.forename: Benoît
authors.initials: B
authors.lastname: Muylkens
mesh_terms: D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002460:Cell Line; D004279:DNA, Viral / Q000235:genetics*; D007242:Herpesvirus 1, Bovine / Q000145:classification / Q000235:genetics* / Q000254:growth & development*; D025881:Herpesvirus 5, Bovine / Q000145:classification / Q000235:genetics* / Q000254:growth & development*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D011995:Recombination, Genetic; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
chemical_list: D004279:DNA, Viral