Generation of recombinant pestiviruses using a full-genome amplification strategy.
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 RT-PCR 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.
Rasmussen
Thomas Bruun
TB
DTU National Veterinary Institute (DTU Vet), Technical University of Denmark, Lindholm, DK-4771 Kalvehave, Denmark. tbrur@vet.dtu.dk
Reimann
Ilona
I
Uttenthal
Ase
A
Leifer
Immanuel
I
Depner
Klaus
K
Schirrmeier
Horst
H
Beer
Martin
M
D000818:Animals; D018083:Border disease virus / Q000235:genetics; D002460:Cell Line; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics; D006692:Classical Swine Fever Virus / Q000235:genetics; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000235:genetics; D030241:Diarrhea Virus 1, Bovine Viral / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D016679:Genome, Viral / Q000235:genetics*; D010576:Pestivirus / Q000235:genetics*; D013552:Swine
D004274:DNA, Recombinant