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The Interrelationship between Promoter Strength, Gene Expression, and Growth Rate

In exponentially growing bacteria, expression of heterologous protein impedes cellular growth rates. Quantitative understanding of the relationship between expression and growth rate will advance our ability to forward engineer bacteria, important for metabolic engineering and synthetic biology appl...

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Autores principales: Bienick, Matthew S., Young, Katherine W., Klesmith, Justin R., Detwiler, Emily E., Tomek, Kyle J., Whitehead, Timothy A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4186888/
https://www.ncbi.nlm.nih.gov/pubmed/25286161
http://dx.doi.org/10.1371/journal.pone.0109105
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author Bienick, Matthew S.
Young, Katherine W.
Klesmith, Justin R.
Detwiler, Emily E.
Tomek, Kyle J.
Whitehead, Timothy A.
author_facet Bienick, Matthew S.
Young, Katherine W.
Klesmith, Justin R.
Detwiler, Emily E.
Tomek, Kyle J.
Whitehead, Timothy A.
author_sort Bienick, Matthew S.
collection PubMed
description In exponentially growing bacteria, expression of heterologous protein impedes cellular growth rates. Quantitative understanding of the relationship between expression and growth rate will advance our ability to forward engineer bacteria, important for metabolic engineering and synthetic biology applications. Recently, a work described a scaling model based on optimal allocation of ribosomes for protein translation. This model quantitatively predicts a linear relationship between microbial growth rate and heterologous protein expression with no free parameters. With the aim of validating this model, we have rigorously quantified the fitness cost of gene expression by using a library of synthetic constitutive promoters to drive expression of two separate proteins (eGFP and amiE) in E. coli in different strains and growth media. In all cases, we demonstrate that the fitness cost is consistent with the previous findings. We expand upon the previous theory by introducing a simple promoter activity model to quantitatively predict how basal promoter strength relates to growth rate and protein expression. We then estimate the amount of protein expression needed to support high flux through a heterologous metabolic pathway and predict the sizable fitness cost associated with enzyme production. This work has broad implications across applied biological sciences because it allows for prediction of the interplay between promoter strength, protein expression, and the resulting cost to microbial growth rates.
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spelling pubmed-41868882014-10-16 The Interrelationship between Promoter Strength, Gene Expression, and Growth Rate Bienick, Matthew S. Young, Katherine W. Klesmith, Justin R. Detwiler, Emily E. Tomek, Kyle J. Whitehead, Timothy A. PLoS One Research Article In exponentially growing bacteria, expression of heterologous protein impedes cellular growth rates. Quantitative understanding of the relationship between expression and growth rate will advance our ability to forward engineer bacteria, important for metabolic engineering and synthetic biology applications. Recently, a work described a scaling model based on optimal allocation of ribosomes for protein translation. This model quantitatively predicts a linear relationship between microbial growth rate and heterologous protein expression with no free parameters. With the aim of validating this model, we have rigorously quantified the fitness cost of gene expression by using a library of synthetic constitutive promoters to drive expression of two separate proteins (eGFP and amiE) in E. coli in different strains and growth media. In all cases, we demonstrate that the fitness cost is consistent with the previous findings. We expand upon the previous theory by introducing a simple promoter activity model to quantitatively predict how basal promoter strength relates to growth rate and protein expression. We then estimate the amount of protein expression needed to support high flux through a heterologous metabolic pathway and predict the sizable fitness cost associated with enzyme production. This work has broad implications across applied biological sciences because it allows for prediction of the interplay between promoter strength, protein expression, and the resulting cost to microbial growth rates. Public Library of Science 2014-10-06 /pmc/articles/PMC4186888/ /pubmed/25286161 http://dx.doi.org/10.1371/journal.pone.0109105 Text en © 2014 Bienick et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Bienick, Matthew S.
Young, Katherine W.
Klesmith, Justin R.
Detwiler, Emily E.
Tomek, Kyle J.
Whitehead, Timothy A.
The Interrelationship between Promoter Strength, Gene Expression, and Growth Rate
title The Interrelationship between Promoter Strength, Gene Expression, and Growth Rate
title_full The Interrelationship between Promoter Strength, Gene Expression, and Growth Rate
title_fullStr The Interrelationship between Promoter Strength, Gene Expression, and Growth Rate
title_full_unstemmed The Interrelationship between Promoter Strength, Gene Expression, and Growth Rate
title_short The Interrelationship between Promoter Strength, Gene Expression, and Growth Rate
title_sort interrelationship between promoter strength, gene expression, and growth rate
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4186888/
https://www.ncbi.nlm.nih.gov/pubmed/25286161
http://dx.doi.org/10.1371/journal.pone.0109105
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