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Journal of Virology, February 2005, p. 2440-2448, Vol. 79, No. 4
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.4.2440-2448.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Characterization of Helper Virus-Independent Cytopathogenic Classical Swine Fever Virus Generated by an In Vivo RNA Recombination System

Andreas Gallei,1 Till Rümenapf,1 Heinz-Jürgen Thiel,1 and Paul Becher1*

Institut für Virologie, Justus-Liebig-Universität, Giessen, Germany1

Received 1 July 2004/ Accepted 28 September 2004

Molecular analyses revealed that most cytopathogenic (cp) pestivirus strains evolve from noncytopathogenic (noncp) viruses by nonhomologous RNA recombination. In contrast to bovine viral diarrhea virus (BVDV), cp classical swine fever virus (CSFV) field isolates were rarely detected and always represented helper virus-dependent subgenomes. To investigate RNA recombination in more detail, we recently established an in vivo system allowing the efficient generation of recombinant cp BVDV strains in cell culture after transfecting a synthetic subgenomic and nonreplicatable transcript into cells being infected with noncp BVDV (A. Gallei, A. Pankraz, H.-J. Thiel, and P. Becher, J. Virol. 78:6271-6281, 2004). Using an analogous approach, the first helper virus-independent cp CSFV strain (CP G1) has now been generated by RNA recombination. Accordingly, this study demonstrates the applicability of RNA recombination for designing new viral RNA genomes. The genomic RNA of CP G1 has a calculated size of 18.139 kb, almost 6 kb larger than all previously described CSFV genomes. It contains cellular sequences encoding a polyubiquitin fragment directly upstream of the nonstructural protein NS3 coding gene together with a duplication of viral sequences. CP G1 induces a cytopathic effect on different tissue culture cell lines from pigs and cattle. Subsequent analyses addressed growth kinetics, expression of NS3, and genetic stability of CP G1.

* Corresponding author. Mailing address: Institut für Virologie, Justus-Liebig-Universität, Giessen, Germany. Phone: 49-641-99-38376. Fax: 49-641-99-38359. E-mail: Paul.Becher{at}vetmed.uni-giessen.de.

Journal of Virology, February 2005, p. 2440-2448, Vol. 79, No. 4
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.4.2440-2448.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Gallei, A., Blome, S., Gilgenbach, S., Tautz, N., Moennig, V., Becher, P. (2008). Cytopathogenicity of Classical Swine Fever Virus Correlates with Attenuation in the Natural Host. J. Virol. 82: 9717-9729 [Abstract] [Full Text]  
  • Gallei, A., Widauer, S., Thiel, H.-J., Becher, P. (2006). Mutations in the palm region of a plus-strand RNA virus polymerase result in attenuated phenotype. J. Gen. Virol. 87: 3631-3636 [Abstract] [Full Text]  
  • Heimann, M., Sosa, G. R., Martoglio, B., Thiel, H.-J., Rumenapf, T. (2006). Core Protein of Pestiviruses Is Processed at the C Terminus by Signal Peptide Peptidase. J. Virol. 80: 1915-1921 [Abstract] [Full Text]  
  • Gallei, A., Orlich, M., Thiel, H.-J., Becher, P. (2005). Noncytopathogenic Pestivirus Strains Generated by Nonhomologous RNA Recombination: Alterations in the NS4A/NS4B Coding Region. J. Virol. 79: 14261-14270 [Abstract] [Full Text]  


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