Heterologous Expression of Membrane Proteins in E. coli

Peer Depping; María Monserrat Román Lara; Athanasios Kesidis; Roslyn M Bill; Alice J Rothnie; Douglas F Browning; Alan D Goddard

doi:10.1007/978-1-0716-2368-8_4

Heterologous Expression of Membrane Proteins in E. coli

Peer Depping, María Monserrat Román Lara, Athanasios Kesidis, Roslyn M Bill, Alice J Rothnie, Douglas F Browning, Alan D Goddard

Research output: Chapter in Book/Published conference output › Chapter (peer-reviewed) › peer-review

Abstract

Over the decades, the bacterium Escherichia coli (E. coli) has become the cornerstone of recombinant protein production, used for heterologous synthesis of a variety of membrane proteins. Due to its rapid growth to high densities in cheap media, and its ease of manipulation and handling, E. coli is an excellent host cell for a range of membrane protein targets. Furthermore, its genetic tractability allows for a variety of gene constructs to be screened for optimal expression conditions, resulting in relatively high yields of membrane protein in a short amount of time. Here, we describe the general workflow for the production of membrane proteins in E. coli. The protocols we provide show how the gene of interest is modified, transferred to an expression vector and host, and how membrane protein yields can be optimized and analyzed. The examples we illustrate are well suited for scientists who are starting their journey into the world of membrane protein production.

Original language	English
Title of host publication	Heterologous Expression of Membrane Proteins
Subtitle of host publication	Methods in Molecular Biology
Editors	I. Mus-Veteau
Publisher	Springer
Pages	59-78
Number of pages	20
Volume	2507
ISBN (Electronic)	978-1-0716-2368-8
ISBN (Print)	978-1-0716-2367-1
DOIs	https://doi.org/10.1007/978-1-0716-2368-8_4
Publication status	Published - 1 Jul 2022

Publication series

Name	Methods in Molecular Biology
Volume	2507
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Bibliographical note

Funding Information:
We are grateful for funding from the European Union’s Horizon 2020 research and innovation programme under Marie Sklodowska-Curie grant agreement No. 847419 (MemTrain) as well as the ERACoBioTech MeMBrane project and BBSRC (BB/R02152X/1) to A.D.G, A.J.R, and R.M.B. D.F.B was supported by BBSRC grants BB/M018261/1 and BB/R017689/1. M.M.R.L. acknowledges support from grant CONACyT (2018-000024-01EXTF-00053).

Keywords

Escherichia coli
Integral membrane proteins
Outer membrane proteins
Recombinant protein production

Access to Document

10.1007/978-1-0716-2368-8_4

Cite this

Depping, P., Román Lara, M. M., Kesidis, A., Bill, R. M., Rothnie, A. J., Browning, D. F., & Goddard, A. D. (2022). Heterologous Expression of Membrane Proteins in E. coli. In I. Mus-Veteau (Ed.), Heterologous Expression of Membrane Proteins: Methods in Molecular Biology (Vol. 2507, pp. 59-78). (Methods in Molecular Biology; Vol. 2507). Springer. https://doi.org/10.1007/978-1-0716-2368-8_4

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Depping, P, Román Lara, MM, Kesidis, A, Bill, RM , Rothnie, AJ , Browning, DF & Goddard, AD 2022, Heterologous Expression of Membrane Proteins in E. coli. in I Mus-Veteau (ed.), Heterologous Expression of Membrane Proteins: Methods in Molecular Biology. vol. 2507, Methods in Molecular Biology, vol. 2507, Springer, pp. 59-78. https://doi.org/10.1007/978-1-0716-2368-8_4

Heterologous Expression of Membrane Proteins in E. coli. / Depping, Peer; Román Lara, María Monserrat; Kesidis, Athanasios et al.
Heterologous Expression of Membrane Proteins: Methods in Molecular Biology. ed. / I. Mus-Veteau. Vol. 2507 Springer, 2022. p. 59-78 (Methods in Molecular Biology; Vol. 2507).

Research output: Chapter in Book/Published conference output › Chapter (peer-reviewed) › peer-review

TY - CHAP

T1 - Heterologous Expression of Membrane Proteins in E. coli

AU - Depping, Peer

AU - Román Lara, María Monserrat

AU - Kesidis, Athanasios

AU - Bill, Roslyn M

AU - Rothnie, Alice J

AU - Browning, Douglas F

AU - Goddard, Alan D

N1 - Funding Information: We are grateful for funding from the European Union’s Horizon 2020 research and innovation programme under Marie Sklodowska-Curie grant agreement No. 847419 (MemTrain) as well as the ERACoBioTech MeMBrane project and BBSRC (BB/R02152X/1) to A.D.G, A.J.R, and R.M.B. D.F.B was supported by BBSRC grants BB/M018261/1 and BB/R017689/1. M.M.R.L. acknowledges support from grant CONACyT (2018-000024-01EXTF-00053).

PY - 2022/7/1

Y1 - 2022/7/1

N2 - Over the decades, the bacterium Escherichia coli (E. coli) has become the cornerstone of recombinant protein production, used for heterologous synthesis of a variety of membrane proteins. Due to its rapid growth to high densities in cheap media, and its ease of manipulation and handling, E. coli is an excellent host cell for a range of membrane protein targets. Furthermore, its genetic tractability allows for a variety of gene constructs to be screened for optimal expression conditions, resulting in relatively high yields of membrane protein in a short amount of time. Here, we describe the general workflow for the production of membrane proteins in E. coli. The protocols we provide show how the gene of interest is modified, transferred to an expression vector and host, and how membrane protein yields can be optimized and analyzed. The examples we illustrate are well suited for scientists who are starting their journey into the world of membrane protein production.

AB - Over the decades, the bacterium Escherichia coli (E. coli) has become the cornerstone of recombinant protein production, used for heterologous synthesis of a variety of membrane proteins. Due to its rapid growth to high densities in cheap media, and its ease of manipulation and handling, E. coli is an excellent host cell for a range of membrane protein targets. Furthermore, its genetic tractability allows for a variety of gene constructs to be screened for optimal expression conditions, resulting in relatively high yields of membrane protein in a short amount of time. Here, we describe the general workflow for the production of membrane proteins in E. coli. The protocols we provide show how the gene of interest is modified, transferred to an expression vector and host, and how membrane protein yields can be optimized and analyzed. The examples we illustrate are well suited for scientists who are starting their journey into the world of membrane protein production.

KW - Escherichia coli

KW - Integral membrane proteins

KW - Outer membrane proteins

KW - Recombinant protein production

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DO - 10.1007/978-1-0716-2368-8_4

M3 - Chapter (peer-reviewed)

C2 - 35773577

SN - 978-1-0716-2367-1

VL - 2507

T3 - Methods in Molecular Biology

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EP - 78

BT - Heterologous Expression of Membrane Proteins

A2 - Mus-Veteau, I.

PB - Springer

ER -

Heterologous Expression of Membrane Proteins in E. coli

Abstract

Publication series

Bibliographical note

Keywords

Access to Document

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