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

Virology. 2016 Oct;497:157-162. doi: 10.1016/j.virol.2016.07.015. Epub 2016 Jul 26.

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.

MeSH terms

  • Animals
  • Chlorocebus aethiops
  • Cloning, Molecular
  • Gene Expression*
  • Gene Order
  • Genes, Reporter*
  • Genome, Viral*
  • Green Fluorescent Proteins / genetics
  • Recombination, Genetic*
  • Vero Cells
  • Zika Virus / genetics*
  • Zika Virus Infection / virology

Substances

  • Green Fluorescent Proteins