Synthesis of functional GABAA receptors in stable insect cell lines.
We have synthesised the beta 1-subunit of the bovine GABAA receptor in stable, continuous insect (Spodoptera frugiperda) cell lines. A cDNA was integrated randomly into the insect cell genome under control of a baculovirus immediate early (IE-1) gene promoter. Transformed cells were obtained by co-transfection of the insect cells with pIEK1.GR beta 1, encoding the beta 1 subunit cDNA, and pIEK1.neo, encoding the neomycin resistance gene. G-418-resistant clones were selected and expanded into continuous cell lines synthesising functional, GABA-gated, homo-oligomeric chloride channels. These cell lines had significant advantages over the transient baculovirus expression system for the characterisation of receptors using electrophysiological recording techniques.
Joyce
K A
KA
School of Biological and Molecular Sciences, Oxford Brookes University, UK.
Atkinson
A E
AE
Bermudez
I
I
Beadle
D J
DJ
King
L A
LA
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001640:Bicuculline / Q000494:pharmacology; D002417:Cattle; D002460:Cell Line; D018118:Chloride Channels / Q000502:physiology; D018076:DNA, Complementary / Q000235:genetics; D004351:Drug Resistance / Q000235:genetics; D004594:Electrophysiology; D017781:Genes, Immediate-Early; D015640:Ion Channel Gating / Q000187:drug effects; D009036:Moths; D009355:Neomycin; D010957:Plasmids; D011401:Promoter Regions, Genetic; D018079:Receptors, GABA / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology; D011994:Recombinant Proteins / Q000096:biosynthesis; D014162:Transfection; D005680:gamma-Aminobutyric Acid / Q000494:pharmacology
D018118:Chloride Channels; D018076:DNA, Complementary; D018079:Receptors, GABA; D011994:Recombinant Proteins; D005680:gamma-Aminobutyric Acid; D009355:Neomycin; D001640:Bicuculline