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Heterologous Gene Expression in E.coli

Volume 705 of the series Methods in Molecular Biology pp 15-30

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SUMO Fusion Technology for Enhanced Protein Expression and Purification in Prokaryotes and Eukaryotes

  • Raymond J. Peroutka IIIAffiliated withLifeSensors Inc.
  • , Steven J. OrcuttAffiliated withLifeSensors Inc.
  • , James E. StricklerAffiliated withLifeSensors Inc.
  • , Tauseef R. ButtAffiliated withLifeSensors Inc. Email author 

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Abstract

The preparation of sufficient amounts of high-quality protein samples is the major bottleneck for structural proteomics. The use of recombinant proteins has increased significantly during the past decades. The most commonly used host, Escherichia coli, presents many challenges including protein misfolding, protein degradation, and low solubility. A novel SUMO fusion technology appears to enhance protein expression and solubility (http://​www.​lifesensors.​com). Efficient removal of the SUMO tag by SUMO protease in vitro facilitates the generation of target protein with a native N-terminus. In addition to its physiological relevance in eukaryotes, SUMO can be used as a powerful biotechnology tool for enhanced functional protein expression in prokaryotes and eukaryotes.

Key words

SUMO Smt3 SUMO protease1 protein expression protein solubility protein purification SUMOstar SUMOstar protease