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  • Title: Direct recovery of infectious pestivirus from a full-length RT-PCR amplicon.
    Author: Rasmussen TB, Reimann I, Hoffmann B, Depner K, Uttenthal A, Beer M.
    Journal: J Virol Methods; 2008 May; 149(2):330-3. PubMed ID: 18336923.
    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.
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