Optimizing E. coli-based membrane protein production using Lemo21(DE3) or pReX and GFP-Fusions

Grietje Kuipers, Markus Peschke, Nurzian Bernsel Ismail, Anna Hjelm, Susan Schlegel, David Vikström, Joen Luirink, Jan Willem de Gier

Research output: Chapter in Book / Report / Conference proceedingChapterAcademicpeer-review

Abstract

Optimizing the conditions for the production of membrane proteins in E. coli is usually a laborious and time-consuming process. Combining the Lemo21(DE3) strain or the pReX T7-based expression vector with membrane proteins C-terminally fused to Green Fluorescent Protein (GFP) greatly facilitates the optimization of membrane protein production yields. Both Lemo21(DE3) and pReX allow precise regulation of expression intensities of genes encoding membrane proteins, which is critical to identify the optimal production condition for a membrane protein. The use of GFP-fusions allows direct monitoring and visualization of membrane proteins at any stage during the production optimization process.

LanguageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc
Pages109-126
Number of pages18
Volume1586
DOIs
StatePublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1586
ISSN (Print)10643745

Fingerprint

Green Fluorescent Proteins
Membrane Proteins
Escherichia coli
Gene Expression Regulation
Protein C

Keywords

  • E. coli
  • Fluorescence
  • GFP
  • Lemo21(DE3)
  • Membrane protein
  • pReX
  • Production

Cite this

Kuipers, G., Peschke, M., Ismail, N. B., Hjelm, A., Schlegel, S., Vikström, D., ... de Gier, J. W. (2017). Optimizing E. coli-based membrane protein production using Lemo21(DE3) or pReX and GFP-Fusions. In Methods in Molecular Biology (Vol. 1586, pp. 109-126). (Methods in Molecular Biology; Vol. 1586). Humana Press Inc. DOI: 10.1007/978-1-4939-6887-9_7
Kuipers, Grietje ; Peschke, Markus ; Ismail, Nurzian Bernsel ; Hjelm, Anna ; Schlegel, Susan ; Vikström, David ; Luirink, Joen ; de Gier, Jan Willem. / Optimizing E. coli-based membrane protein production using Lemo21(DE3) or pReX and GFP-Fusions. Methods in Molecular Biology. Vol. 1586 Humana Press Inc, 2017. pp. 109-126 (Methods in Molecular Biology).
@inbook{972f06d7ddb742bf8fe82098df7bb500,
title = "Optimizing E. coli-based membrane protein production using Lemo21(DE3) or pReX and GFP-Fusions",
abstract = "Optimizing the conditions for the production of membrane proteins in E. coli is usually a laborious and time-consuming process. Combining the Lemo21(DE3) strain or the pReX T7-based expression vector with membrane proteins C-terminally fused to Green Fluorescent Protein (GFP) greatly facilitates the optimization of membrane protein production yields. Both Lemo21(DE3) and pReX allow precise regulation of expression intensities of genes encoding membrane proteins, which is critical to identify the optimal production condition for a membrane protein. The use of GFP-fusions allows direct monitoring and visualization of membrane proteins at any stage during the production optimization process.",
keywords = "E. coli, Fluorescence, GFP, Lemo21(DE3), Membrane protein, pReX, Production",
author = "Grietje Kuipers and Markus Peschke and Ismail, {Nurzian Bernsel} and Anna Hjelm and Susan Schlegel and David Vikstr{\"o}m and Joen Luirink and {de Gier}, {Jan Willem}",
year = "2017",
doi = "10.1007/978-1-4939-6887-9_7",
language = "English",
volume = "1586",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc",
pages = "109--126",
booktitle = "Methods in Molecular Biology",

}

Kuipers, G, Peschke, M, Ismail, NB, Hjelm, A, Schlegel, S, Vikström, D, Luirink, J & de Gier, JW 2017, Optimizing E. coli-based membrane protein production using Lemo21(DE3) or pReX and GFP-Fusions. in Methods in Molecular Biology. vol. 1586, Methods in Molecular Biology, vol. 1586, Humana Press Inc, pp. 109-126. DOI: 10.1007/978-1-4939-6887-9_7

Optimizing E. coli-based membrane protein production using Lemo21(DE3) or pReX and GFP-Fusions. / Kuipers, Grietje; Peschke, Markus; Ismail, Nurzian Bernsel; Hjelm, Anna; Schlegel, Susan; Vikström, David; Luirink, Joen; de Gier, Jan Willem.

Methods in Molecular Biology. Vol. 1586 Humana Press Inc, 2017. p. 109-126 (Methods in Molecular Biology; Vol. 1586).

Research output: Chapter in Book / Report / Conference proceedingChapterAcademicpeer-review

TY - CHAP

T1 - Optimizing E. coli-based membrane protein production using Lemo21(DE3) or pReX and GFP-Fusions

AU - Kuipers,Grietje

AU - Peschke,Markus

AU - Ismail,Nurzian Bernsel

AU - Hjelm,Anna

AU - Schlegel,Susan

AU - Vikström,David

AU - Luirink,Joen

AU - de Gier,Jan Willem

PY - 2017

Y1 - 2017

N2 - Optimizing the conditions for the production of membrane proteins in E. coli is usually a laborious and time-consuming process. Combining the Lemo21(DE3) strain or the pReX T7-based expression vector with membrane proteins C-terminally fused to Green Fluorescent Protein (GFP) greatly facilitates the optimization of membrane protein production yields. Both Lemo21(DE3) and pReX allow precise regulation of expression intensities of genes encoding membrane proteins, which is critical to identify the optimal production condition for a membrane protein. The use of GFP-fusions allows direct monitoring and visualization of membrane proteins at any stage during the production optimization process.

AB - Optimizing the conditions for the production of membrane proteins in E. coli is usually a laborious and time-consuming process. Combining the Lemo21(DE3) strain or the pReX T7-based expression vector with membrane proteins C-terminally fused to Green Fluorescent Protein (GFP) greatly facilitates the optimization of membrane protein production yields. Both Lemo21(DE3) and pReX allow precise regulation of expression intensities of genes encoding membrane proteins, which is critical to identify the optimal production condition for a membrane protein. The use of GFP-fusions allows direct monitoring and visualization of membrane proteins at any stage during the production optimization process.

KW - E. coli

KW - Fluorescence

KW - GFP

KW - Lemo21(DE3)

KW - Membrane protein

KW - pReX

KW - Production

UR - http://www.scopus.com/inward/record.url?scp=85018406678&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85018406678&partnerID=8YFLogxK

U2 - 10.1007/978-1-4939-6887-9_7

DO - 10.1007/978-1-4939-6887-9_7

M3 - Chapter

VL - 1586

T3 - Methods in Molecular Biology

SP - 109

EP - 126

BT - Methods in Molecular Biology

PB - Humana Press Inc

ER -

Kuipers G, Peschke M, Ismail NB, Hjelm A, Schlegel S, Vikström D et al. Optimizing E. coli-based membrane protein production using Lemo21(DE3) or pReX and GFP-Fusions. In Methods in Molecular Biology. Vol. 1586. Humana Press Inc. 2017. p. 109-126. (Methods in Molecular Biology). Available from, DOI: 10.1007/978-1-4939-6887-9_7