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Title: Evaluation of Gaussia luciferase and foot-and-mouth disease virus 2A translational interrupter chimeras as polycistronic reporters for transgene expression

Journal Article · · BMC Biotechnology (Online)
ORCiD logo [1];  [2];  [2];  [2]
  1. Plum Island Animal Disease Center, Greenport, NY (United States). U.S. Department of Homeland Security Science and Technology Directorate; Plum Island Animal Disease Center Research Participation Program (PIADC), Oak Ridge, TN (United States). Oak Ridge Institute for Science and Education
  2. Plum Island Animal Disease Center, Greenport, NY (United States). U.S. Department of Homeland Security Science and Technology Directorate
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Background: The Gaussia princeps luciferase is used as a stand-alone reporter of transgene expression for in vitro and in vivo expression systems due to the rapid and easy monitoring of luciferase activity. We sought to simultaneously quantitate production of other recombinant proteins by transcriptionally linking the Gaussia princeps luciferase gene to other genes of interest through the foot-and-mouth disease virus 2A translational interrupter sequence. Results: We produced six plasmids, each encoding a single open reading frame, with the foot-and-mouth disease virus 2A sequence placed either N-terminal or C-terminal to the Gaussia princeps luciferase gene. Two plasmids included novel Gaussia princeps luciferase variants with the position 1 methionine deleted. Placing a foot-and-mouth disease virus 2A translational interrupter sequence on either the N- or C-terminus of the Gaussia princeps luciferase gene did not prevent the secretion or luminescence of resulting chimeric luciferase proteins. We also measured the ability of another polycistronic plasmid vector with a 2A-luciferase sequence placed downstream of the foot-and-mouth disease virus P1 and 3C protease genes to produce of foot-and-mouth disease virus-like particles and luciferase activity from transfected cells. Incorporation of the 2A-luciferase sequence into a transgene encoding foot-and-mouth disease virus structural proteins retained luciferase activity and the ability to form virus-like particles. Conclusions: We demonstrated a mechanism for the near real-time, sequential, non-destructive quantitative monitoring of transcriptionally-linked recombinant proteins and a valuable method for monitoring transgene expression in recombinant vaccine constructs.

Research Organization:
Oak Ridge Institute for Science and Education (ORISE), Oak Ridge, TN (United States)
Sponsoring Organization:
USDOE
Grant/Contract Number:
AC05-06OR23100
OSTI ID:
1494693
Journal Information:
BMC Biotechnology (Online), Vol. 17, Issue 1; ISSN 1472-6750
Publisher:
BioMed CentralCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 9 works
Citation information provided by
Web of Science

References (21)

Reactions of Intracellular Crystals of Foot-and-Mouth Disease Virus with Ferritin-tagged Antibody journal April 1969
Analysis of the aphthovirus 2A/2B polyprotein ‘cleavage’ mechanism indicates not a proteolytic reaction, but a novel translational effect: a putative ribosomal ‘skip’ journal May 2001
Secreted Gaussia luciferase as a sensitive reporter gene for in vivo and ex vivo studies of airway gene transfer journal April 2011
Protein Coexpression Using FMDV 2A: Effect of “Linker” Residues journal January 2013
Codon-Optimized Gaussia Luciferase cDNA for Mammalian Gene Expression in Culture and in Vivo journal March 2005
Secreted Luciferase for In Vivo Evaluation of Systemic Protein Delivery in Mice journal March 2012
A secreted luciferase for ex vivo monitoring of in vivo processes journal January 2008
Gaussia luciferase reporter assay for monitoring biological processes in culture and in vivo journal April 2009
Correction for LaRocco et al., A Continuous Bovine Kidney Cell Line Constitutively Expressing Bovine α V β 6 Integrin Has Increased Susceptibility to Foot-and-Mouth Disease Virus journal January 2015
The cleavage activities of aphthovirus and cardiovirus 2A proteins. journal January 1997
Analysis of Foot-and-Mouth Disease Virus Type O1 Brugge Neutralization Epitopes Using Monoclonal Antibodies journal October 1986
Secreted Gaussia Luciferase as a Biomarker for Monitoring Tumor Progression and Treatment Response of Systemic Metastases journal December 2009
Occurrence, function and evolutionary origins of '2A-like' sequences in virus genomes journal April 2008
Recombinant Gaussia Luciferase. Overexpression, Purification, and Analytical Application of a Bioluminescent Reporter for DNA Hybridization journal September 2002
Polycistronic Viral Vectors journal September 2002
Production and characterization of two serotype independent monoclonal antibodies against foot-and-mouth disease virus journal January 2007
Superluminescent Variants of Marine Luciferases for Bioassays journal November 2011
Foot-and-mouth disease virus 2A oligopeptide mediated cleavage of an artificial polyprotein. journal February 1994
Electron microscopic observation of crystalline arrays of foot-and-mouth disease virus. journal January 1966
A Continuous Bovine Kidney Cell Line Constitutively Expressing Bovine  V 6 Integrin Has Increased Susceptibility to Foot-and-Mouth Disease Virus journal March 2013
Gaussia Luciferase for Bioluminescence Tumor Monitoring in Comparison with Firefly Luciferase journal September 2011

Cited By (1)

Foot-and-Mouth Disease (FMD) Virus 3C Protease Mutant L127P: Implications for FMD Vaccine Development journal September 2017

Figures / Tables (8)

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Table 1(p. 6)table Table 1

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59 BASIC BIOLOGICAL SCIENCES
Gaussia luciferase
Foot-and-mouth disease virus
2A
Bicistronic
Polycistronic
Biomarker
Virus-like particles