J Mol Genet Med 2010, Vol 4, 239-246 doi: jmgm Published online: 02 September 2010 Full Text: (HTML | PDF ~260kb | refs) A seven plasmid-based system for the rescue of influenza C virus Karin Pachler, Juliane Mayr and Reinhard Vlasak* Department of Molecular Biology, University of Salzburg, Billrothstrasse 11, 5020 Salzburg, Austria *Correspondence to: Reinhard Vlasak, Email: reinhard.vlasak@sbg.ac.at, Tel: +43 662 8044 7330, Fax: +43 662 8044 7209 Received: 07 June 2010, Revised: 02 August 2010, Accepted: 10 August 2010 ----------------------------------------------------------------------------------------------------------------- ABSTRACT We report the establishment of a reverse-genetics system for the rescue of recombinant influenza C/JJ/50 virus from seven plasmids. The nucleotide sequence of the whole C/JJ/50 genome was determined and full-length cDNAs were cloned into an RNA pol I/pol II-based bidirectional vector. Transfection of Vero cells and subsequent amplification on MDCK cells yielded viral HA titres of 128. The utility of this bidirectional approach was proved by generating a reassortant virus encoding the NS segment from strain C/JHB/1/66 and a virus with mutations in the noncoding ends of PB1. The latter virus, which has a base-pair mutation within the proposed double-stranded region of the PB1 termini, exhibited impaired replication. In conclusion, our efficient seven-plasmid system for the rescue of recombinant influenza C virus may be used to study the influenza C virus life cycle in more detail and for generation of influenza C virus-based vectors. KEYWORDS: Influenza C virus, reverse genetics, reassortant virus, noncoding ends, panhandle/corkscrew RNA model ----------------------------------------------------------------------------------------------------------------- Privacy | Disclaimer |
