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Classical swine fever virus: recovery of infectious viruses from cDNA constructs and generation of recombinant cytopathogenic defective interfering particles

Meyers, G.; Thiel, H.J.; Rümenapf, T.

Journal of Virology 70(3): 1588-1595

1996


ISSN/ISBN: 0022-538X
PMID: 8627678
Accession: 002778936

The 5'- and 3'-terminal sequences of the genomic RNA from classical swine fever virus (CSFV) were determined, and the resulting information was used for construction of full-length CSFV cDNA clones. After transfection of in vitro-transcribed RNA derived from a cDNA construct, infectious CSFV was recovered from porcine cells. To confirm the de novo generation of infectious CSFV from cloned DNA, a genetically tagged CSFV was constructed. In comparison with parental CSFV, the recombinant viruses were retarded in growth by about 1 order of magnitude. Introduction of a deletion by exchange of part of the full-length construct for corresponding cDNA fragments derived from the genomes of cytopathogenic CSFV defective interfering particles (DIs) (G. Meyers and H.J. Thiel, J. Virol. 69:3683-3689, 1995) resulted in recovery of cytopathogenic DIs from in vitro-transcribed RNA. This finding confirmed the hypothesis that the deletion previously identified in the DI genomes is responsible for their cytopathogenicity. The established system will allow novel approaches to analysis of pestiviral molecular biology and in particular to elucidation of the molecular basis of attenuation and cytopathogenicity of these viruses.

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