Cell-free expression has emerged as a new standard for the production of membrane proteins. The reduction of expression complexity in cell-free systems eliminates central bottlenecks and allows the reliable and efficient synthesis of many different types of membrane proteins. Furthermore, the open accessibility of cell-free reactions enables the co-translational solubilization of cell-free expressed membrane proteins in a large variety of supplied additives. Hydrophobic environments can therefore be adjusted according to the requirements of individual membrane protein targets. We present different approaches for the preparative scale cell-free production of G-protein-coupled receptors using the extracts of Escherichia coli cells. We exemplify expression conditions implementing detergents, nanodiscs, or liposomes. The generated protein samples could be directly used for further functional characterization.
CITATION STYLE
Orbán, E., Proverbio, D., Haberstock, S., Dötsch, V., & Bernhard, F. (2015). Cell-free expression of G-protein-coupled receptors. Methods in Molecular Biology (Clifton, N.J.), 1261, 171–195. https://doi.org/10.1007/978-1-4939-2230-7_10
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