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Methods Mol Biol. 2014;1091:161-78. doi: 10.1007/978-1-62703-691-7_11.

Cell-free protein synthesis for functional and structural studies.

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Department of Biochemistry, University of Wisconsin-Madison, Madison, WI, USA.


Recent advances in cell-free protein expression systems have made them reliable and practical for functional and structural studies of a wide variety of proteins. In particular, wheat germ cell-free translation can consistently produce target proteins in microgram quantities from relatively inexpensive, small-scale reactions. Here we describe our small-scale protein expression method for rapidly producing proteins for functional assay and techniques for determining if the target is suitable for scale-up to amounts potentially needed for structure determination. The cell-free system is versatile and can be easily customized with the inclusion of additives. We describe simple modifications used for producing membrane proteins.

[Indexed for MEDLINE]

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