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Inexpensive protein overexpression driven by the NarL transcription activator protein

Most Escherichia coli overexpression vectors used for recombinant protein production (RPP) depend on organic inducers, for example, sugars or simple conjugates. However, these can be expensive and, sometimes, chemically unstable. To simplify this and to cut the cost of RPP, we have developed vectors...

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Autores principales: Hothersall, Joanne, Lai, Sandie, Zhang, Nan, Godfrey, Rita E., Ruanto, Patcharawarin, Bischoff, Sarah, Robinson, Colin, Overton, Tim W., Busby, Stephen J. W., Browning, Douglas F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9314961/
https://www.ncbi.nlm.nih.gov/pubmed/35211956
http://dx.doi.org/10.1002/bit.28071
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author Hothersall, Joanne
Lai, Sandie
Zhang, Nan
Godfrey, Rita E.
Ruanto, Patcharawarin
Bischoff, Sarah
Robinson, Colin
Overton, Tim W.
Busby, Stephen J. W.
Browning, Douglas F.
author_facet Hothersall, Joanne
Lai, Sandie
Zhang, Nan
Godfrey, Rita E.
Ruanto, Patcharawarin
Bischoff, Sarah
Robinson, Colin
Overton, Tim W.
Busby, Stephen J. W.
Browning, Douglas F.
author_sort Hothersall, Joanne
collection PubMed
description Most Escherichia coli overexpression vectors used for recombinant protein production (RPP) depend on organic inducers, for example, sugars or simple conjugates. However, these can be expensive and, sometimes, chemically unstable. To simplify this and to cut the cost of RPP, we have developed vectors controlled by the Escherichia coli nitrate‐responsive NarL transcription activator protein, which use nitrate, a cheap, stable, and abundant inorganic ion, to induce high‐level controlled RPP. We show that target proteins, such as green fluorescent protein, human growth hormone, and single‐chain variable region antibody fragments can be expressed to high levels using our promoter systems. As nitrate levels are high in many commercial fertilizers, we demonstrate that controlled RPP can be achieved using readily available and inexpensive garden products.
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spelling pubmed-93149612022-07-30 Inexpensive protein overexpression driven by the NarL transcription activator protein Hothersall, Joanne Lai, Sandie Zhang, Nan Godfrey, Rita E. Ruanto, Patcharawarin Bischoff, Sarah Robinson, Colin Overton, Tim W. Busby, Stephen J. W. Browning, Douglas F. Biotechnol Bioeng ARTICLES Most Escherichia coli overexpression vectors used for recombinant protein production (RPP) depend on organic inducers, for example, sugars or simple conjugates. However, these can be expensive and, sometimes, chemically unstable. To simplify this and to cut the cost of RPP, we have developed vectors controlled by the Escherichia coli nitrate‐responsive NarL transcription activator protein, which use nitrate, a cheap, stable, and abundant inorganic ion, to induce high‐level controlled RPP. We show that target proteins, such as green fluorescent protein, human growth hormone, and single‐chain variable region antibody fragments can be expressed to high levels using our promoter systems. As nitrate levels are high in many commercial fertilizers, we demonstrate that controlled RPP can be achieved using readily available and inexpensive garden products. John Wiley and Sons Inc. 2022-03-06 2022-06 /pmc/articles/PMC9314961/ /pubmed/35211956 http://dx.doi.org/10.1002/bit.28071 Text en © 2022 The Authors. Biotechnology and Bioengineering published by Wiley Periodicals LLC https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle ARTICLES
Hothersall, Joanne
Lai, Sandie
Zhang, Nan
Godfrey, Rita E.
Ruanto, Patcharawarin
Bischoff, Sarah
Robinson, Colin
Overton, Tim W.
Busby, Stephen J. W.
Browning, Douglas F.
Inexpensive protein overexpression driven by the NarL transcription activator protein
title Inexpensive protein overexpression driven by the NarL transcription activator protein
title_full Inexpensive protein overexpression driven by the NarL transcription activator protein
title_fullStr Inexpensive protein overexpression driven by the NarL transcription activator protein
title_full_unstemmed Inexpensive protein overexpression driven by the NarL transcription activator protein
title_short Inexpensive protein overexpression driven by the NarL transcription activator protein
title_sort inexpensive protein overexpression driven by the narl transcription activator protein
topic ARTICLES
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9314961/
https://www.ncbi.nlm.nih.gov/pubmed/35211956
http://dx.doi.org/10.1002/bit.28071
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