Journal
VETERINARY MICROBIOLOGY
Volume 142, Issue 1-2, Pages 13-17Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.vetmic.2009.09.037
Keywords
Pestivirus; Flaviviridae; Long RT-PCR; Full-genome amplicon; Infectious clone; pBeloBAC11
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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. (C) 2009 Elsevier B.V. All rights reserved.
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