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Predictable, Tunable Protein Production in Salmonella for Studying Host-Pathogen Interactions

Here we describe the use of synthetic genetic elements to improve the predictability and tunability of episomal protein production in Salmonella. We used a multi-pronged approach, in which a series of variable-strength synthetic promoters were combined with a synthetic transcriptional terminator, an...

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Autores principales: Cooper, Kendal G., Chong, Audrey, Starr, Tregei, Finn, Ciaran E., Steele-Mortimer, Olivia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5696353/
https://www.ncbi.nlm.nih.gov/pubmed/29201859
http://dx.doi.org/10.3389/fcimb.2017.00475
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author Cooper, Kendal G.
Chong, Audrey
Starr, Tregei
Finn, Ciaran E.
Steele-Mortimer, Olivia
author_facet Cooper, Kendal G.
Chong, Audrey
Starr, Tregei
Finn, Ciaran E.
Steele-Mortimer, Olivia
author_sort Cooper, Kendal G.
collection PubMed
description Here we describe the use of synthetic genetic elements to improve the predictability and tunability of episomal protein production in Salmonella. We used a multi-pronged approach, in which a series of variable-strength synthetic promoters were combined with a synthetic transcriptional terminator, and plasmid copy number variation. This yielded a series of plasmids that drive uniform production of fluorescent and endogenous proteins, over a wide dynamic range. We describe several examples where this system is used to fine-tune constitutive expression in Salmonella, providing an efficient means to titrate out toxic effects of protein production.
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spelling pubmed-56963532017-11-30 Predictable, Tunable Protein Production in Salmonella for Studying Host-Pathogen Interactions Cooper, Kendal G. Chong, Audrey Starr, Tregei Finn, Ciaran E. Steele-Mortimer, Olivia Front Cell Infect Microbiol Microbiology Here we describe the use of synthetic genetic elements to improve the predictability and tunability of episomal protein production in Salmonella. We used a multi-pronged approach, in which a series of variable-strength synthetic promoters were combined with a synthetic transcriptional terminator, and plasmid copy number variation. This yielded a series of plasmids that drive uniform production of fluorescent and endogenous proteins, over a wide dynamic range. We describe several examples where this system is used to fine-tune constitutive expression in Salmonella, providing an efficient means to titrate out toxic effects of protein production. Frontiers Media S.A. 2017-11-16 /pmc/articles/PMC5696353/ /pubmed/29201859 http://dx.doi.org/10.3389/fcimb.2017.00475 Text en Copyright © 2017 Cooper, Chong, Starr, Finn and Steele-Mortimer. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Cooper, Kendal G.
Chong, Audrey
Starr, Tregei
Finn, Ciaran E.
Steele-Mortimer, Olivia
Predictable, Tunable Protein Production in Salmonella for Studying Host-Pathogen Interactions
title Predictable, Tunable Protein Production in Salmonella for Studying Host-Pathogen Interactions
title_full Predictable, Tunable Protein Production in Salmonella for Studying Host-Pathogen Interactions
title_fullStr Predictable, Tunable Protein Production in Salmonella for Studying Host-Pathogen Interactions
title_full_unstemmed Predictable, Tunable Protein Production in Salmonella for Studying Host-Pathogen Interactions
title_short Predictable, Tunable Protein Production in Salmonella for Studying Host-Pathogen Interactions
title_sort predictable, tunable protein production in salmonella for studying host-pathogen interactions
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5696353/
https://www.ncbi.nlm.nih.gov/pubmed/29201859
http://dx.doi.org/10.3389/fcimb.2017.00475
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