1. Strategies for the expression of SUMO-modified target proteins in Escherichia coli.
- Author
-
Saitoh H, Uwada J, and Azusa K
- Subjects
- Algorithms, Cloning, Molecular methods, Gene Expression, Genetic Vectors genetics, Genetic Vectors metabolism, Microbiological Techniques, Models, Biological, Protein Processing, Post-Translational, Proteins metabolism, Recombinant Fusion Proteins isolation & purification, Recombinant Fusion Proteins metabolism, Small Ubiquitin-Related Modifier Proteins metabolism, Transformation, Bacterial physiology, Clinical Laboratory Techniques, Escherichia coli genetics, Proteins genetics, Recombinant Fusion Proteins genetics, Small Ubiquitin-Related Modifier Proteins genetics
- Abstract
We previously described the establishment of a binary vector system that allows co-expression of SUMO conjugation enzymes and a target protein of interest, leading to efficient SUMO modification and the production of a large amount of recombinant SUMO-modified proteins in Escherichia coli. The advantages of this E. coli expression/modification approach include scalability of experiments, low cost, fast growth, and a lack of proteases that cleave the isopeptide linkage between SUMO and the target protein. Thus, this E. coli method provides a useful alternative to authentic SUMO modification assays, such as in vitro SUMO conjugation and in vivo SUMO modification using baculovirus or mammalian cell culture, that are usually complicated, time-consuming and expensive.
- Published
- 2009
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