Evaluation of Gaussia luciferase and foot-and-mouth disease virus 2A translational interrupter chimeras as polycistronic reporters for transgene expression
Abstract
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 abilitymore »
- Authors:
-
- 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
- Plum Island Animal Disease Center, Greenport, NY (United States). U.S. Department of Homeland Security Science and Technology Directorate
- Publication Date:
- Research Org.:
- Oak Ridge Institute for Science and Education (ORISE), Oak Ridge, TN (United States)
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 1494693
- Grant/Contract Number:
- AC05-06OR23100
- Resource Type:
- Accepted Manuscript
- Journal Name:
- BMC Biotechnology (Online)
- Additional Journal Information:
- Journal Name: BMC Biotechnology (Online); Journal Volume: 17; Journal Issue: 1; Journal ID: ISSN 1472-6750
- Publisher:
- BioMed Central
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; Gaussia luciferase; Foot-and-mouth disease virus; 2A; Bicistronic; Polycistronic; Biomarker; Virus-like particles
Citation Formats
Puckette, Michael, Burrage, Thomas, Neilan, John G., and Rasmussen, Max. Evaluation of Gaussia luciferase and foot-and-mouth disease virus 2A translational interrupter chimeras as polycistronic reporters for transgene expression. United States: N. p., 2017.
Web. doi:10.1186/s12896-017-0367-0.
Puckette, Michael, Burrage, Thomas, Neilan, John G., & Rasmussen, Max. Evaluation of Gaussia luciferase and foot-and-mouth disease virus 2A translational interrupter chimeras as polycistronic reporters for transgene expression. United States. https://doi.org/10.1186/s12896-017-0367-0
Puckette, Michael, Burrage, Thomas, Neilan, John G., and Rasmussen, Max. Mon .
"Evaluation of Gaussia luciferase and foot-and-mouth disease virus 2A translational interrupter chimeras as polycistronic reporters for transgene expression". United States. https://doi.org/10.1186/s12896-017-0367-0. https://www.osti.gov/servlets/purl/1494693.
@article{osti_1494693,
title = {Evaluation of Gaussia luciferase and foot-and-mouth disease virus 2A translational interrupter chimeras as polycistronic reporters for transgene expression},
author = {Puckette, Michael and Burrage, Thomas and Neilan, John G. and Rasmussen, Max},
abstractNote = {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.},
doi = {10.1186/s12896-017-0367-0},
journal = {BMC Biotechnology (Online)},
number = 1,
volume = 17,
place = {United States},
year = {Mon Jun 12 00:00:00 EDT 2017},
month = {Mon Jun 12 00:00:00 EDT 2017}
}
Web of Science
Figures / Tables:
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Production and characterization of two serotype independent monoclonal antibodies against foot-and-mouth disease virus
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A secreted luciferase for ex vivo monitoring of in vivo processes
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Gaussia luciferase reporter assay for monitoring biological processes in culture and in vivo
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Reactions of Intracellular Crystals of Foot-and-Mouth Disease Virus with Ferritin-tagged Antibody
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Analysis of Foot-and-Mouth Disease Virus Type O1 Brugge Neutralization Epitopes Using Monoclonal Antibodies
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- Journal of General Virology, Vol. 82, Issue 5
A Continuous Bovine Kidney Cell Line Constitutively Expressing Bovine V 6 Integrin Has Increased Susceptibility to Foot-and-Mouth Disease Virus
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- Journal of Clinical Microbiology, Vol. 51, Issue 6
Secreted Gaussia Luciferase as a Biomarker for Monitoring Tumor Progression and Treatment Response of Systemic Metastases
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Works referencing / citing this record:
Foot-and-Mouth Disease (FMD) Virus 3C Protease Mutant L127P: Implications for FMD Vaccine Development
journal, September 2017
- Puckette, Michael; Clark, Benjamin A.; Smith, Justin D.
- Journal of Virology, Vol. 91, Issue 22
Foot-and-Mouth Disease (FMD) Virus 3C Protease Mutant L127P: Implications for FMD Vaccine Development
journal, September 2017
- Puckette, Michael; Clark, Benjamin A.; Smith, Justin D.
- Journal of Virology, Vol. 91, Issue 22
Figures / Tables found in this record: