|
KMID : 1161519990030020181
|
|
Animal Cells and Systems
1999 Volume.3 No. 2 p.181 ~ p.185
|
|
|
Improved method for heterologous expression of ion channels in Xenopus Oocyte: A PCR shortcut to Oocyte expression
|
|
Kim Han-Seop
Lee Chang-Ho
Moon Eun-Pyo
Min Churl-K.
|
|
|
Abstract
|
|
|
|
Xenopus oocyte is one of the widely used heterologous expression systems of ion channels for electrophysiological studies. Here we describe a new method in which cRNA produced by polymerase chain reaction (PCR) and in vitro transcription is injected to express ion channels in oocytes. This method enables us (1) to eliminate all or a part of the untranslated region of the cDNA and to replace it with a known sequence which helps increase the expression level in oocytes, and (2) to use the PCR product for in vitro transcription without subcloning. Using this method, the expression level of one of the neuronal nicotinic acetyfcholine receptors (nAChRs) α7 subtype in oocytes was systematically increased by more than 100?fold, which was confirmed both by the α?Bungarotoxin (αBgt) binding assay and the current measurement
|
|
|
KEYWORD
|
|
|
Xenopus oocyte, Heterologous expression, cRNA, Nuronal nicotinic receptor α7 subtype
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
 
|