Abstract
Cell-free protein synthesis is advantageous for the expression of protein complexes, since it is suitable for the co-expression of two or more components of the target protein complexes. The quantity and the quality of cell-free expressed complexes are generally better than those of protein complexes expressed in conventional cell-based systems, because various parameters, such as the stoichiometry of the component proteins, can be more precisely controlled. In this chapter, we describe techniques for the expression of protein complexes by an Escherichia coli cell-free protein synthesis system, which has been successfully utilized in crystallographic structural studies.
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Acknowledgements
We thank T. Mishima and T. Nakayama for their assistance in manuscript preparation. This work was supported by the RIKEN Structural Genomics/Proteomics Initiative (RSGI), the National Project on Protein Structural and Functional Analyses, and the Targeted Proteins Research Program (TPRP), of the Ministry of Education, Culture, Sports, Science and Technology of Japan.
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Terada, T., Murata, T., Shirouzu, M., Yokoyama, S. (2014). Cell-Free Expression of Protein Complexes for Structural Biology. In: Chen, Y. (eds) Structural Genomics. Methods in Molecular Biology, vol 1091. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-691-7_10
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DOI: https://doi.org/10.1007/978-1-62703-691-7_10
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