Rescue and characterization of recombinant virus from a new world zika virus infectious clone

James Weger-Lucarelli, Nisha K. Duggal, Aaron Brault, Brian J. Geiss, Gregory D. Ebel

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Infectious cDNA clones allow for genetic manipulation of a virus, thus facilitating work on vaccines, pathogenesis, replication, transmission and viral evolution. Here we describe the construction of an infectious clone for Zika virus (ZIKV), which is currently causing an explosive outbreak in the Americas. To prevent toxicity to bacteria that is commonly observed with flavivirus-derived plasmids, we generated a two-plasmid system which separates the genome at the NS1 gene and is more stable than full-length constructs that could not be successfully recovered without mutations. After digestion and ligation to join the two fragments, full-length viral RNA can be generated by in vitro transcription with T7 RNA polymerase. Following electroporation of transcribed RNA into cells, virus was recovered that exhibited similar in vitro growth kinetics and in vivo virulence and infection phenotypes in mice and mosquitoes, respectively.

Original languageEnglish (US)
Article numbere55857
JournalJournal of Visualized Experiments
Volume2017
Issue number124
DOIs
StatePublished - Jun 7 2017
Externally publishedYes

Fingerprint

RNA
Viruses
Plasmids
Clone Cells
Flavivirus
Electroporation
Viral RNA
Genes
Culicidae
Disease Outbreaks
Ligation
Virulence
Digestion
Vaccines
Complementary DNA
Growth kinetics
Genome
Transcription
Bacteria
Phenotype

Keywords

  • Arbovirus
  • CDNA clone
  • Flavivirus
  • Immunology
  • Infectious clone
  • Issue 124
  • Molecular clone
  • Zika virus
  • ZIKV

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Rescue and characterization of recombinant virus from a new world zika virus infectious clone. / Weger-Lucarelli, James; Duggal, Nisha K.; Brault, Aaron; Geiss, Brian J.; Ebel, Gregory D.

In: Journal of Visualized Experiments, Vol. 2017, No. 124, e55857, 07.06.2017.

Research output: Contribution to journalArticle

Weger-Lucarelli, James ; Duggal, Nisha K. ; Brault, Aaron ; Geiss, Brian J. ; Ebel, Gregory D. / Rescue and characterization of recombinant virus from a new world zika virus infectious clone. In: Journal of Visualized Experiments. 2017 ; Vol. 2017, No. 124.
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