Journal article
Insertion of a GFP Reporter Gene in Influenza Virus
Current protocols in microbiology (Online), Vol.15(1), pp.15G.4.1-15G.4.16
2013
DOI: 10.1002/9780471729259.mc15g04s29
PMCID: PMC3878617
PMID: 23686828
Abstract
The incorporation of a fluorescent reporter gene into a replication competent influenza A virus (IAV) has made it possible to trace IAV infection
in vivo.
This protocol describes the process of inserting a green fluorescent protein (GFP) reporter into the IAV genome using the established reverse genetics system. The strategy begins with the reorganization of segment eight of the IAV genome, during which the open reading frames of non-structural protein 1 (NS1) and the nuclear export protein (NEP) are separated to allow for GFP fusion to the NS1 protein. The NS1, GFP, and NEP open reading frames (ORF) are then cloned into the IAV rescue system backbone. Upon construction of the GFP encoding segment eight rescue plasmid, recombinant NS1-GFP influenza virus can be rescued via co-transfection with the remaining seven rescue plasmids. The generated NS1-GFP IAV can subsequently be used to visualize infected cells both
in vitro
and
in vivo.
Details
- Title: Subtitle
- Insertion of a GFP Reporter Gene in Influenza Virus
- Creators
- Jasmine T Perez - Department of Microbiology, University of Chicago, Chicago, IllinoisAdolfo García-Sastre - Department of Microbiology, Mount Sinai School of Medicine, New York, New YorkBalaji Manicassamy - Department of Microbiology, University of Chicago, Chicago, Illinois
- Resource Type
- Journal article
- Publication Details
- Current protocols in microbiology (Online), Vol.15(1), pp.15G.4.1-15G.4.16
- DOI
- 10.1002/9780471729259.mc15g04s29
- PMID
- 23686828
- PMCID
- PMC3878617
- NLM abbreviation
- Curr Protoc Microbiol
- ISSN
- 1934-8525
- eISSN
- 1934-8533
- Grant note
- R00 AI095320 || AI / National Institute of Allergy and Infectious Diseases Extramural Activities : NIAID
- Language
- English
- Date published
- 2013
- Academic Unit
- Microbiology and Immunology
- Record Identifier
- 9984083274302771
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