A stable full-length yellow fever virus cDNA clone and the role of conserved RNA elements in flavivirus replication
- PMID: 12692292
- DOI: 10.1099/vir.0.18860-0
A stable full-length yellow fever virus cDNA clone and the role of conserved RNA elements in flavivirus replication
Abstract
Yellow fever virus (YF) is the prototype member of the Flavivirus genus. Here, we report the successful construction of a full-length infectious cDNA clone of the vaccine strain YF-17D. YF cDNA was cloned into a low-copy-number plasmid backbone and stably maintained in several E. coli strains. Transcribed RNAs had a specific infectivity of 10(5)-10(6) p.f.u. ( micro g RNA)(-1), and the resulting virus exhibited growth kinetics, plaque morphology and proteolytic processing similar to the parental virus in cell culture. This clone was used to analyse the importance of conserved flavivirus RNA sequences and the 3' stem-loop structure in virus replication. The conserved sequences 5'CS and CS1, as well as the 3' stem-loop structure, were found to be essential for virus replication in cell culture, whereas the conserved sequence CS2 and the region containing YF-specific repeated sequences were dispensable. This infectious clone will aid future studies on YF replication and pathogenesis, as well as facilitate the development of YF-17D-based recombinant vaccines.
Similar articles
-
Transcription of infectious yellow fever RNA from full-length cDNA templates produced by in vitro ligation.New Biol. 1989 Dec;1(3):285-96. New Biol. 1989. PMID: 2487295
-
Characterization of an infectious clone of the wild-type yellow fever virus Asibi strain that is able to infect and disseminate in mosquitoes.J Gen Virol. 2005 Jun;86(Pt 6):1747-1751. doi: 10.1099/vir.0.80746-0. J Gen Virol. 2005. PMID: 15914853
-
Characterization of infectious Murray Valley encephalitis virus derived from a stably cloned genome-length cDNA.J Gen Virol. 1999 Dec;80 ( Pt 12):3115-3125. doi: 10.1099/0022-1317-80-12-3115. J Gen Virol. 1999. PMID: 10567642
-
The yellow fever 17D vaccine virus: molecular basis of viral attenuation and its use as an expression vector.Braz J Med Biol Res. 1997 Feb;30(2):157-68. doi: 10.1590/s0100-879x1997000200002. Braz J Med Biol Res. 1997. PMID: 9239300 Review.
-
Flavivirus reverse genetic systems, construction techniques and applications: a historical perspective.Antiviral Res. 2015 Feb;114:67-85. doi: 10.1016/j.antiviral.2014.12.007. Epub 2014 Dec 12. Antiviral Res. 2015. PMID: 25512228 Free PMC article. Review.
Cited by
-
Reprint of: Core protein-mediated 5'-3' annealing of the West Nile virus genomic RNA in vitro.Virus Res. 2012 Nov;169(2):448-57. doi: 10.1016/j.virusres.2012.09.009. Epub 2012 Sep 27. Virus Res. 2012. PMID: 23022255 Free PMC article.
-
Construction and characterisation of a complete reverse genetics system of dengue virus type 3.Mem Inst Oswaldo Cruz. 2013 Dec;108(8):983-91. doi: 10.1590/0074-0276130298. Mem Inst Oswaldo Cruz. 2013. PMID: 24402142 Free PMC article.
-
Single-stranded positive-sense RNA viruses generated in days using infectious subgenomic amplicons.J Gen Virol. 2014 Nov;95(Pt 11):2462-2467. doi: 10.1099/vir.0.068023-0. Epub 2014 Jul 22. J Gen Virol. 2014. PMID: 25053561 Free PMC article.
-
Structure of the Flavivirus helicase: implications for catalytic activity, protein interactions, and proteolytic processing.J Virol. 2005 Aug;79(16):10268-77. doi: 10.1128/JVI.79.16.10268-10277.2005. J Virol. 2005. PMID: 16051820 Free PMC article.
-
Core protein-mediated 5'-3' annealing of the West Nile virus genomic RNA in vitro.Virus Res. 2012 Aug;167(2):226-35. doi: 10.1016/j.virusres.2012.05.003. Epub 2012 May 28. Virus Res. 2012. PMID: 22652509 Free PMC article.
Publication types
MeSH terms
Substances
Grants and funding
LinkOut - more resources
Full Text Sources
Other Literature Sources
Miscellaneous
