Characterization of heterologously expressed transporter genes by patch- and voltage-clamp methods: Application to cyclic nucleotide-dependent responses

Handle URI:
http://hdl.handle.net/10754/562965
Title:
Characterization of heterologously expressed transporter genes by patch- and voltage-clamp methods: Application to cyclic nucleotide-dependent responses
Authors:
Lemtiri-Chlieh, Fouad ( 0000-0002-7418-2623 ) ; Ali, Rashid Ayesha
Abstract:
The application of patch- and voltage-clamp methods to study ion transport can be limited by many hurdles: the size of the cells to be patched and/or stabbed, the subcellular localization of the molecule of interest, and its density of expression that could be too low even in their own native environment. Functional expression of genes using recombinant DNA technology not only overcomes those hurdles but also affords additional and elegant investigations such as single-point mutation studies and subunit associations/regulations. In this chapter, we give a step-by-step description of two electrophysiological methods, patch clamp and two-electrode voltage clamp (TEVC), that are routinely used in combination with heterologous gene expression to assist researchers interested in the identification and characterization of ion transporters. We describe how to (1) obtain and maintain the cells suitable for the use with each of the above-mentioned methods (i.e., HEK-293 cells and yeast spheroplasts to use with the patch-clamp methodology and Xenopus laevis oocytes with TEVC), (2) transfect/inject them with the gene of interest, and (3) record ion transport activities. © Springer Science+Business Media New York 2013.
KAUST Department:
Biological and Environmental Sciences and Engineering (BESE) Division
Publisher:
Humana Press
Journal:
Methods in Molecular Biology
Issue Date:
3-Sep-2013
DOI:
10.1007/978-1-62703-441-8-6
PubMed ID:
23681573
Type:
Article
ISSN:
10643745
ISBN:
9781627034401
Appears in Collections:
Articles; Biological and Environmental Sciences and Engineering (BESE) Division

Full metadata record

DC FieldValue Language
dc.contributor.authorLemtiri-Chlieh, Fouaden
dc.contributor.authorAli, Rashid Ayeshaen
dc.date.accessioned2015-08-03T11:17:17Zen
dc.date.available2015-08-03T11:17:17Zen
dc.date.issued2013-09-03en
dc.identifier.isbn9781627034401en
dc.identifier.issn10643745en
dc.identifier.pmid23681573en
dc.identifier.doi10.1007/978-1-62703-441-8-6en
dc.identifier.urihttp://hdl.handle.net/10754/562965en
dc.description.abstractThe application of patch- and voltage-clamp methods to study ion transport can be limited by many hurdles: the size of the cells to be patched and/or stabbed, the subcellular localization of the molecule of interest, and its density of expression that could be too low even in their own native environment. Functional expression of genes using recombinant DNA technology not only overcomes those hurdles but also affords additional and elegant investigations such as single-point mutation studies and subunit associations/regulations. In this chapter, we give a step-by-step description of two electrophysiological methods, patch clamp and two-electrode voltage clamp (TEVC), that are routinely used in combination with heterologous gene expression to assist researchers interested in the identification and characterization of ion transporters. We describe how to (1) obtain and maintain the cells suitable for the use with each of the above-mentioned methods (i.e., HEK-293 cells and yeast spheroplasts to use with the patch-clamp methodology and Xenopus laevis oocytes with TEVC), (2) transfect/inject them with the gene of interest, and (3) record ion transport activities. © Springer Science+Business Media New York 2013.en
dc.publisherHumana Pressen
dc.subjectHEK-293en
dc.subjectHeterologous expressionen
dc.subjectPatch clampen
dc.subjectTwo-electrode voltage clampen
dc.subjectXenopus oocytesen
dc.subjectYeast protoplastsen
dc.titleCharacterization of heterologously expressed transporter genes by patch- and voltage-clamp methods: Application to cyclic nucleotide-dependent responsesen
dc.typeArticleen
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Divisionen
dc.identifier.journalMethods in Molecular Biologyen
dc.contributor.institutionCenter for Vascular Biology, University of Connecticut Health Center, Farmington CT, United Statesen
kaust.authorLemtiri-Chlieh, Fouaden

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