Cell-free expression offers an interesting alternative method to produce membrane proteins in high amounts. Elimination of toxicity problems, reduced proteolytic degradation and a nearly unrestricted option to supply potentially beneficial compounds like cofactors, ligands or chaperones into the reaction are general advantages of cell-free expression systems. Furthermore, the membrane proteins may be translated directly into appropriate hydrophobic and membrane-mimetic surrogates, which might offer significant benefits for the functional folding of the synthesized proteins. Cell-free expression is a rapidly developing and highly versatile technique and several systems of both, prokaryotic and eukaryotic origins, have been established. We provide protocols for the cell-free expression of membrane proteins in different modes including their expression as precipitate as well as their direct synthesis into detergent micelles or lipid bilayers.
CITATION STYLE
Reckel, S., Sobhanifar, S., Durst, F., Löhr, F., Shirokov, V. A., Dötsch, V., & Bernhard, F. (2010). Strategies for the cell-free expression of membrane proteins. Methods in Molecular Biology (Clifton, N.J.), 607, 187–212. https://doi.org/10.1007/978-1-60327-331-2_16
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