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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...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2014
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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. |
format | Online Article Text |
id | pubmed-4186888 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
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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