Direct recovery of infectious Pestivirus from a full-length RT-PCR amplicon

Thomas Bruun Rasmussen, Ilona Reimann, Bernd Hoffmann, Klaus Depner, Åse Uttenthal, Martin Beer

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    This study describes the use of a novel and rapid long reverse transcription (RT)-PCR for the generation of infectious full-length cDNA of pestiviruses. To produce rescued viruses, full-length RT-PCR amplicons of 12.3 kb, including a T7-promotor, were transcribed directly in vitro, and the resulting RNA transcripts were electroporated into ovine cells. Infectious virus was obtained after one cell culture passage. The rescued viruses had a phenotype similar to the parental Border Disease virus strain. Therefore, direct generation of infectious pestiviruses from full-length RT-PCR cDNA products could be a valuable instrument for virus rescue, conservation and further characterization
    Original languageEnglish
    JournalJournal of Virological Methods
    Volume149
    Issue number2
    Pages (from-to)330-330
    ISSN0166-0934
    DOIs
    Publication statusPublished - 2008

    Keywords

    • Virus rescue
    • Full-genome RT-PCR
    • Pestivirus
    • Flaviviridae

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