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Virology. 2016 Oct;497:157-62. doi: 10.1016/j.virol.2016.07.015. Epub 2016 Jul 26.

A robust method for the rapid generation of recombinant Zika virus expressing the GFP reporter gene.

Author information

  • 1Université de La Réunion, CNRS UMR 9192, INSERM U1187, IRD UMR 249, Unité Mixte 134 Processus Infectieux Insulaire Tropical (PIMIT), Plateforme Technologique CYROI, 97490 Sainte Clotilde, France.
  • 2Université de La Réunion, CNRS UMR 9192, INSERM U1187, IRD UMR 249, Unité Mixte 134 Processus Infectieux Insulaire Tropical (PIMIT), Plateforme Technologique CYROI, 97490 Sainte Clotilde, France. Electronic address: philippe.despres@univ-reunion.fr.

Abstract

Zika virus (ZIKV) infection is a major public health problem with severe human congenital and neurological anomalies. The screening of anti-ZIKV compounds and neutralizing antibodies needs reliable and rapid virus-based assays. Here, we described a convenient method leading to the rapid production of molecular clones of ZIKV. To generate a molecular clone of ZIKV strain MR766(NIID), the viral genome was directly assembled into Vero cells after introduction of four overlapping synthetic fragments that cover the full-length genomic RNA sequence. Such strategy has allowed the production of a recombinant ZIKV expressing the GFP reporter gene that is stable over two culturing rounds on Vero cells. Our data demonstrate that the ZIKV reporter virus is a very reliable GFP-based tool for analyzing viral growth and measuring the neutralizing antibody as well as rapid screening of antiviral effect of different classes of inhibitors.

KEYWORDS:

Arbovirus; Emerging disease; Flavivirus; GFP reporter; Molecular clones; Recombinant virus; Zika virus

PMID:
27471954
DOI:
10.1016/j.virol.2016.07.015
[PubMed - in process]
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