Generation of recombinant pestiviruses using a full-genome amplification strategy

Thomas Bruun Rasmussen, I. Reimann, Åse Uttenthal, I. Leifer, K. Depner, H. Schirrmeier, M. Beer

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    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 RTPCR 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.
    Original languageEnglish
    JournalVeterinary Microbiology
    Volume142
    Issue number1-2
    Pages (from-to)13-17
    ISSN0378-1135
    DOIs
    Publication statusPublished - 2010
    Event7th Pestivirus Symposium of the European-Society-of-Veterinary-Virology - Uppsala, Sweden
    Duration: 17 Sep 200819 Sep 2008
    Conference number: 7

    Conference

    Conference7th Pestivirus Symposium of the European-Society-of-Veterinary-Virology
    Number7
    CountrySweden
    CityUppsala
    Period17/09/200819/09/2008

    Keywords

    • Infectious clone
    • pBeloBAC11
    • Full-genome amplicon
    • Pestivirus
    • Long RT-PCR
    • Flaviviridae

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