Abstract
Escherichia coli is widely used as an expression system for production of recombinant proteins of prokaryotic and eukaryotic origin. A large body of knowledge has accumulated throughout the last few decades regarding expression of recombinant proteins in E. coli. However, despite this progress, protein production, primarily of eukaryotic origin, still remains a challenge. The biggest obstacle lies in obtaining large amounts of a given protein in a correctly folded form. Several strategies are being used to increase both yield and solubility. These include expression as fusion proteins, co-expression with molecular chaperones, or with a protein partner(s), and the use of multiple constructs for each protein. In this chapter, we focus on strategies for creating expression vectors, as well as on guidelines for improving recombinant protein solubility.
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Acknowledgments
We thank Prof. J. Sussman, Prof. I. Silman, Prof. G. Schreiber and Prof. Yigal Burstein for their continuous support throughout the study. This research was supported by the European Commission Sixth Framework Research and Technological Development Program “SPINE2-COMPLEXES” Project, under contract No. 031220; a grant from the Israel Ministry of Science, Culture, and Sport to the Israel Structural Proteomics Center; the Divadol Foundation; and the Neuman Foundation.
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Peleg, Y., Unger, T. (2012). Resolving Bottlenecks for Recombinant Protein Expression in E. coli . In: Zanders, E. (eds) Chemical Genomics and Proteomics. Methods in Molecular Biology, vol 800. Humana Press. https://doi.org/10.1007/978-1-61779-349-3_12
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DOI: https://doi.org/10.1007/978-1-61779-349-3_12
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