PMID: 9188607Jul 1, 1997Paper

Infectious RNA transcripts from full-length dengue virus type 2 cDNA clones made in yeast

Journal of Virology
S PoloBarry Falgout

Abstract

The dengue virus type 2 genomic RNA was amplified by reverse transcription-PCR and cloned as four cDNA fragments. We could not assemble these four fragments into full-length cDNA in Escherichia coli. The full-length dengue virus cDNA was constructed by homologous recombination in yeast, either as part of a yeast artificial chromosome or in a yeast-E. coli shuttle vector. Full-length cDNA clones were propagated once in E. coli to prepare useful quantities of DNA. In vitro transcription of these clones produced full-length RNA transcripts. Introduction of these transcripts into LLC-MK2 cells produced typical dengue infection, as judged by cytopathic effects and indirect immunofluorescence. Infectivity was sensitive to RNase digestion and was dependent on the presence of cap analog in the transcription reaction mixture. Virus in the medium was passaged on C6-36 cells to produce stocks, and these stocks had titers and plaque morphologies similar to those of the parental dengue virus type 2. Intracellular dengue virus RNA from cells infected with transcript-derived virus contained an introduced BstEII site, proving that infectivity was derived from RNA transcripts and not from contamination with parental dengue virus. Transcript-der...Continue Reading

References

Jan 2, 1992·Gene·T W ChristiansonP Hieter
Jun 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·C J LaiM Bray
Jan 1, 1990·Annual Review of Microbiology·T J ChambersC M Rice
Nov 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·D R SmithD T Moir
Aug 1, 1990·Trends in Genetics : TIG·C HuxleyI Dunham
Nov 1, 1985·Journal of Virology·S Mizutani, R J Colonno
Mar 29, 1994·Proceedings of the National Academy of Sciences of the United States of America·T P Monath
Nov 1, 1984·Proceedings of the National Academy of Sciences of the United States of America·P AhlquistL S Loesch-Fries

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Citations

Aug 17, 2005·Journal of Virology·Carey L MedinAlan L Rothman
Mar 6, 2013·Anais Da Academia Brasileira De Ciências·Sabrina R A QueirozLaura H V G Gil
Sep 27, 2020·MBio·Chengjin YeLuis Martinez-Sobrido
Nov 17, 2001·Journal of Virology·R CasaisP Britton
May 14, 2003·Journal of Virology·Sang-Im YunYoung-Min Lee
May 22, 2002·Journal of Virology·Pei-Yong ShiKristen A Bernard
Apr 24, 2014·Therapeutic Advances in Vaccines·Usa Thisyakorn, Chule Thisyakorn
Apr 26, 2012·Journal of Virology·Melissanne de Wispelaere, Priscilla L Yang
Apr 26, 2013·Journal of Virology·Melissanne de WispelaerePriscilla L Yang
Jun 1, 2014·The Journal of General Virology·Rodion V GorchakovFarooq Nasar
Dec 15, 2010·The Journal of General Virology·Yingfeng LeiAndrew Dayton
Jan 1, 2013·Uirusu·Ryosuke Suzuki, Eiji Konishi
Jul 24, 2014·The Journal of General Virology·Fabien AubryXavier de Lamballerie
Jun 27, 2014·Viruses·Christopher C Stobart, Martin L Moore
May 5, 2020·Nature·Tran Thi Nhu ThaoVolker Thiel
Sep 22, 2015·International Journal of Molecular Sciences·Yangchao DongYingfeng Lei
Apr 16, 2015·International Journal of Molecular Medicine·Yangchao DongZhikai Xu
Jul 23, 2015·Memórias do Instituto Oswaldo Cruz·Jefferson José da Silva SantosLaura Helena Vega Gonzales Gil
Jan 10, 2014·Memórias do Instituto Oswaldo Cruz·Jefferson José da Silva SantosLaura Helena Vega Gonzales Gil
Jan 16, 2015·Anais Da Academia Brasileira De Ciências·Jefferson J S SantosLaura H V G Gil
Mar 24, 2016·PloS One·Dhanasekaran GovindarajanAndrew J Bett
Jan 7, 1998·Journal of Virology·R P Valle, B Falgout

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