title: Development of a reverse genetics system to generate a recombinant Ebola virus Makona expressing a green fluorescent protein.
abstract: Previous studies have demonstrated the potential application of reverse genetics technology in studying a broad range of aspects of viral biology, including gene regulation, protein function, cell entry, and pathogenesis. Here, we describe a highly efficient reverse genetics system used to generate recombinant Ebola virus (EBOV) based on a recent isolate from a human patient infected during the 2014-2015 outbreak in Western Africa. We also rescued a recombinant EBOV expressing a fluorescent reporter protein from a cleaved VP40 protein fusion. Using this virus and an inexpensive method to quantitate the expression of the foreign gene, we demonstrate its potential usefulness as a tool for screening antiviral compounds and measuring neutralizing antibodies.
authors.affiliation: Centers for Disease Control and Prevention, Atlanta, GA, USA. Electronic address: calbarino@cdc.gov.
authors.forename: César G
authors.initials: CG
authors.lastname: Albariño
authors.affiliation: Centers for Disease Control and Prevention, Atlanta, GA, USA.
authors.forename: Lisa
authors.initials: L
authors.lastname: Wiggleton Guerrero
authors.affiliation: Centers for Disease Control and Prevention, Atlanta, GA, USA.
authors.forename: Michael K
authors.initials: MK
authors.lastname: Lo
authors.affiliation: Centers for Disease Control and Prevention, Atlanta, GA, USA.
authors.forename: Stuart T
authors.initials: ST
authors.lastname: Nichol
authors.affiliation: Centers for Disease Control and Prevention, Atlanta, GA, USA.
authors.forename: Jonathan S
authors.initials: JS
authors.lastname: Towner
mesh_terms: D000354:Africa, Western; D057134:Antibodies, Neutralizing / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000302:isolation & purification; D000998:Antiviral Agents / Q000302:isolation & purification; D004353:Drug Evaluation, Preclinical / Q000379:methods; D029043:Ebolavirus / Q000235:genetics* / Q000302:isolation & purification; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D019142:Hemorrhagic Fever, Ebola / Q000821:virology; D006801:Humans; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000235:genetics; D059386:Reverse Genetics / Q000379:methods*; D017422:Sequence Analysis, DNA; D013194:Staining and Labeling
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D000998:Antiviral Agents; D012367:RNA, Viral; D049452:Green Fluorescent Proteins