Characterization of helper virus-independent cytopathogenic classical swine fever virus generated by an in vivo RNA recombination system.
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.
Gallei
Andreas
A
Institut für Virologie, Justus-Liebig-Universität, Giessen, Germany.
Rümenapf
Till
T
Thiel
Heinz-Jürgen
HJ
Becher
Paul
P
D000818:Animals; D002460:Cell Line; D002461:Cell Line, Transformed; D006692:Classical Swine Fever Virus / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D003588:Cytopathogenic Effect, Viral; D020365:RNA Helicases / Q000378:metabolism*; D012367:RNA, Viral / Q000235:genetics*; D011995:Recombination, Genetic; D012269:Ribosomal Proteins / Q000235:genetics / Q000378:metabolism; D014452:Ubiquitins / Q000235:genetics; D017361:Viral Nonstructural Proteins / Q000235:genetics*; D014765:Viral Vaccines / Q000235:genetics / Q000276:immunology; D016662:Virus Integration
D012367:RNA, Viral; D012269:Ribosomal Proteins; D014452:Ubiquitins; D017361:Viral Nonstructural Proteins; D014765:Viral Vaccines; D020365:RNA Helicases