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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. - Highlights: • Recombinant Ebola virus (EBOV) derived from Makona variant was rescued. • New protocol for viral rescue allows 100% efficiency. • Modified EBOV expresses a green fluorescent protein from a VP40-fused protein. • Modified EBOV was tested as tool to screen antiviral compounds and measure neutralizing antibodies.

Authors:
; ; ;
Publication Date:
OSTI Identifier:
22470192
Resource Type:
Journal Article
Journal Name:
Virology
Additional Journal Information:
Journal Volume: 484; Other Information: Copyright (c) 2015 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0042-6822
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; AFRICA; ANTIBODIES; EFFICIENCY; FLUORESCENCE; GENE REGULATION; GENETICS; HUMAN POPULATIONS; PATHOGENESIS; PATIENTS; PROTEINS; SCREENING; VIRUSES

Citation Formats

Albariño, César G., E-mail: calbarino@cdc.gov, Wiggleton Guerrero, Lisa, Lo, Michael K., Nichol, Stuart T., and Towner, Jonathan S. Development of a reverse genetics system to generate a recombinant Ebola virus Makona expressing a green fluorescent protein. United States: N. p., 2015. Web. doi:10.1016/J.VIROL.2015.06.013.
Albariño, César G., E-mail: calbarino@cdc.gov, Wiggleton Guerrero, Lisa, Lo, Michael K., Nichol, Stuart T., & Towner, Jonathan S. Development of a reverse genetics system to generate a recombinant Ebola virus Makona expressing a green fluorescent protein. United States. doi:10.1016/J.VIROL.2015.06.013.
Albariño, César G., E-mail: calbarino@cdc.gov, Wiggleton Guerrero, Lisa, Lo, Michael K., Nichol, Stuart T., and Towner, Jonathan S. Thu . "Development of a reverse genetics system to generate a recombinant Ebola virus Makona expressing a green fluorescent protein". United States. doi:10.1016/J.VIROL.2015.06.013.
@article{osti_22470192,
title = {Development of a reverse genetics system to generate a recombinant Ebola virus Makona expressing a green fluorescent protein},
author = {Albariño, César G., E-mail: calbarino@cdc.gov and Wiggleton Guerrero, Lisa and Lo, Michael K. and Nichol, Stuart T. and Towner, Jonathan S.},
abstractNote = {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. - Highlights: • Recombinant Ebola virus (EBOV) derived from Makona variant was rescued. • New protocol for viral rescue allows 100% efficiency. • Modified EBOV expresses a green fluorescent protein from a VP40-fused protein. • Modified EBOV was tested as tool to screen antiviral compounds and measure neutralizing antibodies.},
doi = {10.1016/J.VIROL.2015.06.013},
journal = {Virology},
issn = {0042-6822},
number = ,
volume = 484,
place = {United States},
year = {2015},
month = {10}
}