Antibodies, A Laboratory Manual, Second Edition, Edited by Edward A. Greenfield



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Preparing GST-Fusion Proteins from Bacteria

(Protocol summary only for purposes of this preview site)

Glutathione-S-transferase (GST) is one of the most widely used protein tags for generating fusion proteins. The GST tag is 26 kDa in size, made of 220 amino acids, and can be expressed in E. coli as a cytoplasmic protein with full enzymatic activity. GST-fusion proteins can be purified using immobilized glutathione on agarose beads. GST can also be cleaved from the protein of interest by inserting a site-specific cleavage site such as thrombin or TEV (tobacco etch virus) between the gene sequences of GST and the fusion protein. The GST-fusion system has several applications in antibody generation, proteinprotein interactions, DNAprotein interactions, and vaccine production.

Antibodies: A Laboratory Manual, Second edition
Antibodies: A Laboratory Manual, Second edition
Antibodies: A Laboratory Manual, Second edition

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