Functional Expression of Type 1 Rat GABA Transporter in Microinjected Xenopus laevis Oocytes
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In this chapter we describe technical aspects and experimental potential of the two electrodes voltage clamp (TEVC) electrophysiological
approach applied to the Xenopus oocyte-expression system.
This technique is addressed to the study of a particular class of expressed proteins, those responsible to drive ion fluxes
through the plasma membrane. In fact the voltage-clamp technique provides the most direct and sensitive measurement of the
functional properties of ion channels and electrogenic transporters, allowing specific ion currents to be recorded under well-defined
voltage conditions and temporal control.
Besides the study of the physiological properties of specific ion channels as well as their pharmacological modulation, further
applications of the TEVC on oocytes include the possibility to introduce single point mutations in the channel construct and
to infer to possible structural aspects and functional involvements of single amino acidic residues. To achieve these results
these technique should be strictly tied to basic molecular biology techniques. Recent advance of this technique in drug discovery
procedures have been briefly enlightened.
Affiliation(s): (2) Laboratory of Cellular and Molecular Physiology, Department of Structural and Functional Biology, University of Insubria, Varese, Italy
(3) Electrophysiology Unit, Newron Pharmaceuticals, Bresso, MI, Italy
(3) Electrophysiology Unit, Newron Pharmaceuticals, Bresso, MI, Italy
Book Title: In Vitro Transcription and Translation Protocols
Series: Methods in Molecular Biology | Volume: 375 | Pub. Date: May-03-2007 | Page Range: 235-255 | DOI: 10.1007/978-1-59745-388-2_12
Subject: Genetics/Genomics
Key Words:
Xenopus laevis oocytes - functional expression - cRNA microinjection - two electrodes voltage clamp - high-throughput screening
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