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Expression of a genomic clone encoding a brain potassium channel in mammalian cells using lipofection

Abstract

A genomic clone encoding a mouse brain K+ channel (MBK1) was isolated, characterized and expressed in COS cells using the lipofection technique. Transfected COS cells expressed voltage-dependent K+ currents that activated within 20 ms at 0 mV and showed less than 10% inactivation during 250 ms depolarizing pulses at 60 mV. Expressed K+ currents were reversibly blocked by 4-aminopyridine and tetraethylammonium, and were moderately sensitive to dendrotoxin, but insensitive to charybdotoxin. Thus MBK1, expressed transiently in a mammalian cell line, exhibits features characteristic of non-inactivating K+ channels with a conspicuous insensitivity to charybdotoxin. Lipofection is, therefore, a valuable strategy for expression of channel proteins in mammalian cells.

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Abbreviations

4-AP:

4 aminopyridine

TEA:

tetraethylammonium

CTX:

charybdotoxin

DTX:

dendrotoxin

V:

applied voltage

Vrev :

reversal potential

I:

current

G:

conductance

MBK1:

mouse brain potassium channel 1

TES:

N-tris[hydroxymethyl]methyl-2-aminoethanesulfonic acid

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Correspondence to: M. Montal.

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Ferroni, S., Planells-Cases, R., Ahmed, C.M.I. et al. Expression of a genomic clone encoding a brain potassium channel in mammalian cells using lipofection. Eur Biophys J 21, 185–191 (1992). https://doi.org/10.1007/BF00196762

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Key words

  • K+ channels
  • Channel clones
  • Lipofection/ transfection
  • Patch clamp