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*

*Corresponding author for this work

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.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc
Pages109-126
Number of pages18
Volume1586
DOIs
Publication statusPublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1586
ISSN (Print)10643745

Keywords

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

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