Development of a novel DNA-launched dengue virus type 2 infectious clone assembled in a bacterial artificial chromosome

Virus Research
Jose A Usme-CiroJuan C Gallego-Gomez

Abstract

Major progress in Dengue virus (DENV) biology has resulted from the use of infectious clones obtained through reverse genetics. The construction of these clones is commonly based on high- or low-copy number plasmids, yeast artificial chromosomes, yeast-Escherichia coli shuttle vectors, and bacterial artificial chromosomes (BACs). Prokaryotic promoters have consistently been used for the transcription of these clones. The goal of this study was to develop a novel DENV infectious clone in a BAC under the control of the cytomegalovirus immediate-early promoter and to generate a virus with the fusion envelope-green fluorescent protein in an attempt to track virus infection. The transfection of Vero cells with a plasmid encoding the DENV infectious clone facilitated the recovery of infectious particles that increased in titer after serial passages in C6/36 cells. The plaque size and syncytia phenotypes of the recombinant virus were similar to those of the parental virus. Despite the observation of autonomous replication and the detection of low levels of viral genome after two passages, the insertion of green fluorescent protein and Renilla luciferase reporter genes negatively impacted virus rescue. To the best of our knowledge, thi...Continue Reading

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Citations

Jun 27, 2014·Viruses·Christopher C Stobart, Martin L Moore
Dec 17, 2015·The Journal of General Virology·Teera PoyomtipSarin Chimnaronk
May 17, 2015·The Journal of General Virology·Juggragarn JengarnAnan Jongkaewwattana
Jul 23, 2015·Memórias do Instituto Oswaldo Cruz·Jefferson José da Silva SantosLaura Helena Vega Gonzales Gil
Nov 18, 2015·Antimicrobial Agents and Chemotherapy·Hong-Tao XuMark A Wainberg
Nov 6, 2018·Viruses·Ginés Ávila-PérezLuis Martínez-Sobrido
Aug 20, 2019·Antiviral Research·Jing Kai LeeJustin Jang Hann Chu

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Datasets Mentioned

BETA
EU854293

Methods Mentioned

BETA
in
transfection
PCR
Assay
light microscopy
confocal microscopy
in vitro transcription

Software Mentioned

ImagePro Plus
Xcellence Pro
Primer Express

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