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PMID: 8243667 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Synthesis of functional GABAA receptors in stable insect cell lines.

FEBS letters ·Vol. 335 ·No. 1 ·1993-11-29 ·Pages 61-4

Joyce KA, Atkinson AE, Bermudez I, Beadle DJ, King LA

Abstract

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.

MeSH Terms
Animals Baculoviridae/genetics Bicuculline/pharmacology Cattle Cell Line Chloride Channels/physiology DNA, Complementary/genetics Drug Resistance/genetics Electrophysiology Genes, Immediate-Early Ion Channel Gating/drug effects Moths Neomycin Plasmids Promoter Regions, Genetic Receptors, GABA/biosynthesis,genetics,physiology Recombinant Proteins/biosynthesis Transfection gamma-Aminobutyric Acid/pharmacology
Chemicals
Chloride Channels DNA, Complementary Receptors, GABA Recombinant Proteins gamma-Aminobutyric Acid Neomycin Bicuculline
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Joyce K A
School of Biological and Molecular Sciences, Oxford Brookes University, UK.
Atkinson A E
Bermudez I
Beadle D J
King L A
Article Info
Journal
FEBS letters
Abbr.
FEBS Lett
ISSN
0014-5793
Published
1993-11-29
Pages
61-4
Language
English
Region
England
NLM ID
0155157
Subset
IM
Grants
Wellcome Trust · United Kingdom
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