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 outputChapter (peer-reviewed)peer-review


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 languageEnglish
Title of host publicationHeterologous Expression of Membrane Proteins
Subtitle of host publication Methods in Molecular Biology
EditorsI. Mus-Veteau
Number of pages20
ISBN (Electronic)978-1-0716-2368-8
ISBN (Print)978-1-0716-2367-1
Publication statusPublished - 1 Jul 2022

Publication series

NameMethods in Molecular Biology
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).


  • Escherichia coli
  • Integral membrane proteins
  • Outer membrane proteins
  • Recombinant protein production


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