Animal cells and systems

The Korean Society for Integrative Biology (한국통합생물학회)
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Improved Method for Heterologous Expression of Ion Channels in Xenopus Oocyte: a PCR Shortcut to Oocyte Expression

  • Han-Seop Kim (Department of Biological Sciences, College of Natural Sciences, Ajou University) ;
  • Changho Lee (Department of Pharmacology, College of Medicine, Hanyang University) ;
  • Eunpyo Moon (Department of Biological Sciences, College of Natural Sciences, Ajou University) ;
  • Churl K. Min (Department of Biological Sciences, College of Natural Sciences, Ajou University)
  • Published : 1999.06.01
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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 acetylcholine receptors (nAChRs) $\alpha$$_{6}$ subtype in oocytes was systematically increased by more than 100-fold, which was confirmed both by the $\alpha$-Bungarotoxin ($\alpha$,/TEX>Bgt) binding assay and the current measurement.t.

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References

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