GST融合蛋白的准备 Preparation of Glutathione

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Margret B. Einarson and Elena N. Pugacheva
Foxx Chase Cancer Center, Philadelphia, PA 19111
Jason R. Orlinck
Brigham & Women"s Hospital, Boston, MA 02115
Excerpted from Protein: Protein Interactions, Second Edition
Edited by Erica A. Golemis and Peter D. Adams

ABSTRACT
Glutathione-S-Transferase (GST) fusion proteins have a wide range of applications. This protocol is designed for IPTG-inducible bacterial expression vectors.

MATERIALS
Buffers, Solutions, and Reagents

Reduced glutathione, 20 mM prepared in 50 mM Tris-HCl (pH 8.0) Isopropyl-β-D-thio-galactoside (IPTG) LB broth, containing appropriate antibiotic selection Lysis buffer: PBS + 1% Triton X-100, supplemented just before use with protease inhibitors (e.g, final concentrations of 2 µg/ml aprotinin , 1 µg/ml leupeptin , and 25 µg/ml phenylmethylsulfonyl fluoride