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Curr Protoc Protein Sci. 2001 May;Chapter 6:Unit 6.6. doi: 10.1002/0471140864.ps0606s09.

Expression and purification of GST fusion proteins.

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The Wistar Institute, Philadelphia, Pennsylvania, USA.


An increasingly common strategy for expressing proteins and large peptides in prokaryotic systems is to express the protein of interest connected to a "tag" that provides the basis for rapid high-affinity purification. This unit describes the expression and purification of fusion proteins containing the 26-kDa glutathione-S-transferase protein as well as methods for cleaving the affinity tag and repurifying the target protein. Advantages of this popular fusion protein system include high protein yields, high-affinity one-step protein purification of the fusion protein, existence of several alternative protease cleavage sites for removing the affinity tag when required, and ease of removal of the cleaved affinity tag.

[Indexed for MEDLINE]

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