Expression of a genomic clone encoding a brain potassium channel in mammalian cells using lipofection.
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.
Ferroni
S
S
Department of Biology, University of California San Diego, La Jolla 92093-0319.
Planells-Cases
R
R
Ahmed
C M
CM
Montal
M
M
D000818:Animals; D001921:Brain / Q000502:physiology*; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004594:Electrophysiology; D016678:Genome; D015640:Ion Channel Gating / Q000502:physiology; D008564:Membrane Potentials / Q000502:physiology; D051379:Mice; D008807:Mice, Inbred BALB C; D011188:Potassium / Q000494:pharmacology; D015221:Potassium Channels / Q000235:genetics* / Q000502:physiology; D012964:Sodium / Q000494:pharmacology; D014162:Transfection
D015221:Potassium Channels; D004247:DNA; D012964:Sodium; D011188:Potassium