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A modified method for the purification of active large enzymes using the glutathione S-transferase expression system.
- Source :
-
Analytical biochemistry [Anal Biochem] 2012 Feb 15; Vol. 421 (2), pp. 805-7. Date of Electronic Publication: 2011 Dec 13. - Publication Year :
- 2012
-
Abstract
- The glutathione S-transferase (GST) fusion protein system is widely used for high-level expression and efficient purification of recombinant proteins from bacteria. However many GST-tagged proteins are insoluble, and the existing procedures, which employ a mixture of detergents to solubilize the molecules, frequently compromise their functional activity. A further limitation is that large proteins (>80 kDa) are poorly isolated by the current methods and are contaminated by truncated forms. To overcome these problems, we provide here an improved method for efficient purification of active large GST-tagged enzymes such as the 180-kDa GST-fused mitochondrial RNA polymerase.<br /> (Copyright © 2011 Elsevier Inc. All rights reserved.)
- Subjects :
- DNA-Directed RNA Polymerases biosynthesis
DNA-Directed RNA Polymerases genetics
Electrophoresis, Polyacrylamide Gel
Glutathione Transferase biosynthesis
Glutathione Transferase genetics
Humans
Recombinant Fusion Proteins biosynthesis
Recombinant Fusion Proteins genetics
DNA-Directed RNA Polymerases isolation & purification
Glutathione Transferase isolation & purification
Recombinant Fusion Proteins isolation & purification
Subjects
Details
- Language :
- English
- ISSN :
- 1096-0309
- Volume :
- 421
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Analytical biochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 22209735
- Full Text :
- https://doi.org/10.1016/j.ab.2011.12.015