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PMID: 17951760 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Review

Ion-channel reconstitution.

Methods in molecular biology (Clifton, N.J.) ·Vol. 400 ·2007-00-00 ·Pages 571-85

Morera FJ, Vargas G, González C, Rosenmann E, Latorre R

Abstract

In this chapter, a detailed protocol is given for ion-channel reconstitution in the two most used model membranes: planar bilayers and liposomes. In the planar bilayer section, methods are described for the expression of ion channels in Xenopus laevis oocytes, the isolation of their membranes, the insertion of ion channels into the bilayer by vesicle fusion, and the recording of single-ion channel current measurements at a constant applied voltage. The reconstitution of bacterial channels in liposomes is also given. It includes the expression and purification of bacterial channels in E. Coli host strain XL1-blue, the insertion of the channels in liposomes, and the recording of their currents by patch clamping.

MeSH Terms
Animals Escherichia coli/genetics Female Humans Ion Channels/chemistry,genetics,isolation & purification Lipid Bilayers/chemistry Liposomes/chemistry Membrane Potentials Models, Biological Oocytes/chemistry Patch-Clamp Techniques Recombinant Proteins/chemistry,genetics,isolation & purification Xenopus laevis
Chemicals
Ion Channels Lipid Bilayers Liposomes Recombinant Proteins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Morera Francisco J
Centro de Estudios Cientificos, Valdivia, Chile.
Vargas Guillermo
González Carlos
Rosenmann Eduardo
Latorre Ramon
Article Info
Journal
Methods in molecular biology (Clifton, N.J.)
Abbr.
Methods Mol Biol
ISSN
1064-3745
Published
2007-00-00
Pages
571-85
Language
English
Region
United States
NLM ID
9214969
Subset
IM
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