Development of a reverse genetics system to generate a recombinant Ebola virus Makona expressing a green fluorescent protein

Virology. 2015 Oct:484:259-264. doi: 10.1016/j.virol.2015.06.013. Epub 2015 Jun 27.

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.

MeSH terms

  • Africa, Western
  • Antibodies, Neutralizing / isolation & purification
  • Antibodies, Viral / isolation & purification
  • Antiviral Agents / isolation & purification
  • Drug Evaluation, Preclinical / methods
  • Ebolavirus / genetics*
  • Ebolavirus / isolation & purification
  • Genes, Reporter
  • Green Fluorescent Proteins / biosynthesis*
  • Green Fluorescent Proteins / genetics
  • Hemorrhagic Fever, Ebola / virology
  • Humans
  • Molecular Sequence Data
  • RNA, Viral / genetics
  • Reverse Genetics / methods*
  • Sequence Analysis, DNA
  • Staining and Labeling

Substances

  • Antibodies, Neutralizing
  • Antibodies, Viral
  • Antiviral Agents
  • RNA, Viral
  • Green Fluorescent Proteins

Associated data

  • GENBANK/KP178538
  • GENBANK/KR781608
  • GENBANK/KR781609