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

• Published in: European biophysics journal Vol. 21; no. 3; pp. 185 - 191
• Main Authors: FERRONI, S, PLANELLS-CASES, R, IQBAL AHMED, C. M, MONTAL, M
• Format: Journal Article
• Language: English
• Published: Berlin Springer 01.07.1992
• Subjects: Animals; Biological and medical sciences; Brain - physiology; Cell physiology; Cells, Cultured; Cloning, Molecular; DNA - genetics; Electrophysiology; Fundamental and applied biological sciences. Psychology; Genome; Ion Channel Gating - physiology; Membrane Potentials - physiology; Mice; Mice, Inbred BALB C; Molecular and cellular biology; Potassium - pharmacology; Potassium Channels - genetics; Potassium Channels - physiology; Sodium - pharmacology; Transfection; Rodentia; Ionic channel; Gene expression; Characterization; Proteins; Vertebrata; Mammalia; Transfection; Mouse; Isolation; Potassium; Clone; Brain (vertebrata)
• ISSN: 0175-7571; 1432-1017
• DOI: 10.1007/BF00196762

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.