title: A robust method for the rapid generation of recombinant Zika virus expressing the GFP reporter gene.
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.
authors.affiliation: Université 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.
authors.forename: Gilles
authors.initials: G
authors.lastname: Gadea
authors.affiliation: Université 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.
authors.forename: Sandra
authors.initials: S
authors.lastname: Bos
authors.affiliation: Université 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.
authors.forename: Pascale
authors.initials: P
authors.lastname: Krejbich-Trotot
authors.affiliation: Université 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.
authors.forename: Elodie
authors.initials: E
authors.lastname: Clain
authors.affiliation: Université 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.
authors.forename: Wildriss
authors.initials: W
authors.lastname: Viranaicken
authors.affiliation: Université 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.
authors.forename: Chaker
authors.initials: C
authors.lastname: El-Kalamouni
authors.affiliation: Université 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.
authors.forename: Patrick
authors.initials: P
authors.lastname: Mavingui
authors.affiliation: Université 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.
authors.forename: Philippe
authors.initials: P
authors.lastname: Desprès
mesh_terms: D000818:Animals; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D015870:Gene Expression; D023061:Gene Order; D017930:Genes, Reporter; D016679:Genome, Viral; D049452:Green Fluorescent Proteins / Q000235:genetics; D011995:Recombination, Genetic; D014709:Vero Cells; D000071244:Zika Virus / Q000235:genetics*; D000071243:Zika Virus Infection / Q000821:virology
chemical_list: D049452:Green Fluorescent Proteins
keywords: Arbovirus; Emerging disease; Flavivirus; GFP reporter; Molecular clones; Recombinant virus; Zika virus