Ion-channel reconstitution.
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.
Morera
Francisco J
FJ
Centro de Estudios Cientificos, Valdivia, Chile.
Vargas
Guillermo
G
González
Carlos
C
Rosenmann
Eduardo
E
Latorre
Ramon
R
D000818:Animals; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D006801:Humans; D007473:Ion Channels / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D008051:Lipid Bilayers / Q000737:chemistry*; D008081:Liposomes / Q000737:chemistry*; D008564:Membrane Potentials; D008954:Models, Biological; D009865:Oocytes / Q000737:chemistry; D018408:Patch-Clamp Techniques; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D014982:Xenopus laevis
D007473:Ion Channels; D008051:Lipid Bilayers; D008081:Liposomes; D011994:Recombinant Proteins