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Tagging and Fusion Proteins

Tagging and Fusion Proteins

Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...

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Related Experiment Video

Updated: Apr 20, 2026

Orthogonal Protein Purification Facilitated by a Small Bispecific Affinity Tag

Orthogonal Protein Purification Facilitated by a Small Bispecific Affinity Tag

Published on: January 16, 2012

Cleavable self-aggregating tags (cSAT) for protein expression and purification.

Zhanglin Lin¹, Qing Zhao, Bihong Zhou

¹Department of Chemical Engineering, Tsinghua University, One Tsinghua Garden Road, Beijing, 100084, China, zhanglinlin@mail.tsinghua.edu.cn.

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

|December 3, 2014

Summary

A new method uses cleavable self-aggregating tags (cSAT) with Escherichia coli for rapid recombinant protein expression and purification. This technique efficiently produces proteins in active aggregates, simplifying downstream processing and achieving high purity.

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Last Updated: Apr 20, 2026

Orthogonal Protein Purification Facilitated by a Small Bispecific Affinity Tag

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Published on: January 16, 2012

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Non-chromatographic Purification of Recombinant Elastin-like Polypeptides and their Fusions with Peptides and Proteins from Escherichia coli

Non-chromatographic Purification of Recombinant Elastin-like Polypeptides and their Fusions with Peptides and Proteins from Escherichia coli

Published on: June 9, 2014

Area of Science:

Biochemistry
Molecular Biology
Protein Engineering

Background:

The rapid identification of new proteins necessitates efficient expression and purification technologies.
Current methods for recombinant protein production can be time-consuming and complex.

Purpose of the Study:

To develop a simple and rapid scheme for recombinant protein expression and purification.
To utilize self-aggregating peptide fusion tags to drive target proteins into active aggregates in vivo.

Main Methods:

Fusion of target proteins to a self-cleavable Mxe GyrA intein, followed by self-aggregating tags (18A or ELK16).
Expression in Escherichia coli, leading to formation of active protein aggregates.
Separation of aggregates by centrifugation after cell lysis.
Intein self-cleavage using dithiothreitol (DTT) to release the target protein into solution.

Main Results:

The cleavable self-aggregating tags (cSAT) facilitate rapid protein production.
Proteins are driven into active aggregates, simplifying initial separation.
Intein self-cleavage effectively releases the target protein.
Achieved modest purity of around 90% for the intein-ELK16 system.

Conclusions:

The cSAT system offers a quick and efficient method for recombinant protein production.
This approach addresses the critical need for rapid protein expression and purification.
The method is applicable to various target proteins, as demonstrated by application examples.