UNIT 5.22 A Bacterial Cell-Free Expression System to Produce Membrane Proteins and Proteoliposomes: From cDNA to Functional Assay
Published Online: 1 NOV 2008
Copyright © 2008 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Protein Science
How to Cite
Liguori, L., Marques, B. and Lenormand, J.-L. 2008. A Bacterial Cell-Free Expression System to Produce Membrane Proteins and Proteoliposomes: From cDNA to Functional Assay. Current Protocols in Protein Science. 54:5.22:5.22.1–5.22.30.
- Published Online: 1 NOV 2008
- Published Print: NOV 2008
Limitations in the production of folded membrane proteins represent the major bottleneck for functional and structural studies of this huge category of macromolecules. Cell-free expression systems provide an attractive alternative to the classical overexpression systems for producing membrane proteins. However, optimization of these systems remains a challenging task, considering the hydrophobic properties of these molecules. This unit describes the production of eukaryotic membrane proteins either in soluble form or integrated into liposomes using a bacterial cell-free expression system. Liposomes in the reaction mixture induce the direct insertion of freshly produced membrane proteins into the bilayer and allow the formation of functional proteoliposomes in which the membrane proteins are correctly folded. Curr. Protoc. Protein Sci. 54:5.22.1-5.22.30. © 2008 by John Wiley & Sons, Inc.
- cell-free expression system;
- membrane proteins production;
- one-step proteoliposomes synthesis;
- delivery system