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Updated: May 8, 2026

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli

Published on: January 6, 2015

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

Anna Hjelm¹, Susan Schlegel, Thomas Baumgarten

¹Center for Biomembrane Research, Department of Biochemistry and Biophysics, Stockholm University, Stockholm, Sweden.

Methods in Molecular Biology (Clifton, N.J.)

|September 3, 2013

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Optimizing membrane protein overexpression and purification in E. coli is challenging. Using the Lemo21(DE3) strain with Green Fluorescent Protein (GFP) fusions simplifies identifying optimal conditions for high-quality protein production.

Area of Science:

Biochemistry
Molecular Biology
Structural Biology

Background:

Overexpressing and purifying membrane proteins from E. coli is often difficult and slow.
Identifying optimal expression levels is crucial for obtaining sufficient high-quality protein for research.

Purpose of the Study:

To streamline the optimization of membrane protein overexpression and purification in E. coli.
To facilitate the production of high-quality membrane protein material for structural and functional studies.

Main Methods:

Utilized the Lemo21(DE3) bacterial strain for controlled protein expression.
Employed Green Fluorescent Protein (GFP) C-terminal fusions to monitor and optimize protein production.
Developed a simplified protocol for identifying optimal expression conditions.

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Related Experiment Videos

Last Updated: May 8, 2026

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli

Published on: January 6, 2015

Characterization of Membrane Transporters by Heterologous Expression in E. coli and Production of Membrane Vesicles

Characterization of Membrane Transporters by Heterologous Expression in E. coli and Production of Membrane Vesicles

Published on: December 31, 2019

Co-Translational Insertion of Membrane Proteins into Preformed Nanodiscs

Co-Translational Insertion of Membrane Proteins into Preformed Nanodiscs

Published on: November 19, 2020

Main Results:

The Lemo21(DE3) strain enabled efficient identification of optimal expression intensity for membrane proteins.
C-terminal GFP fusions facilitated the assessment of protein production levels.
The combined approach significantly reduced the time and effort required for membrane protein production.

Conclusions:

Combining the Lemo21(DE3) strain and GFP fusions offers a robust strategy for optimizing membrane protein overexpression and purification.
This method accelerates the generation of high-quality membrane proteins for downstream applications in structural and functional biology.