Generation of recombinant pestiviruses using a full-genome amplification strategy

Thomas Bruun Rasmussen , Ilona Reimann , Åse Uttenthal , Immanuel Leifer , Klaus Depner , Horst Schirrmeier , Martin Beer

Complete genome amplification of viral RNA provides a new tool for the generation of modified viruses. We have recently reported a full-genome amplification strategy for recovery of pestiviruses (Rasmussen et al., 2008). A full-length cDNA amplicon corresponding to the Border disease virus-Gifhorn genome was generated by long RT-PCR and then RNA transcripts derived from this amplicon were used to rescue infectious virus. Here, we have now used this full-genome amplification strategy for efficient and robust amplification of three additional pestivirus strains: the vaccine strain C and the virulent Paderborn strain of Classical swine fever virus plus the CP7 strain of Bovine viral diarrhoea virus. The amplicons were cloned directly into a stable single-copy bacterial artificial chromosome generating full-length pestivirus DNAs from which infectious RNA transcripts could be also derived.

Mots clés: long RT-PCR; full-genome amplicon; infectious clone; pBeloBAC11

Thomas Bruun Rasmussen, Ilona Reimann, Åse Uttenthal, Immanuel Leifer, Klaus Depner, et al.. Generation of recombinant pestiviruses using a full-genome amplification strategy. Veterinary Microbiology, 2010, 142 (1-2), pp.13. ⟨10.1016/j.vetmic.2009.09.037⟩. ⟨hal-00578396⟩