Review
doi: 10.1007/978-1-59745-388-2_3.
Cell-free production of integral membrane proteins on a preparative scale
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- PMID: 17634596
- DOI: 10.1007/978-1-59745-388-2_3
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Review
Cell-free production of integral membrane proteins on a preparative scale
Methods Mol Biol.
2007.
Abstract
The chapter will focus on the high level cell-free production of integral membrane proteins having multiple transmembrane segments by using an individual coupled transcription/translation system based on an Escherichia coli S30-extract. We describe in detail the setup and optimization of the cell-free expression technique to obtain the maximum yield of recombinant proteins. The protocol can be used for the expression of soluble membrane proteins as well as for their production as a precipitate. In addition, we will provide protocols for the efficient solubilization and reconstitution of membrane proteins directly from the cell-free produced precipitates.
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