The production of a stably transformed insect cell line expressing an invertebrate GABAa receptor βsubunit

L A Smith; M Amar; Robert J Harvey; M G Darlison; F G P Earley; D J Beadle; L A King; I Bermudez

doi:10.3109/10799899509045205

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The production of a stably transformed insect cell line expressing an invertebrate GABAa receptor βsubunit

Journal article

Peer reviewed

The production of a stably transformed insect cell line expressing an invertebrate GABAa receptor βsubunit

L A Smith, M Amar, Robert J Harvey, M G Darlison, F G P Earley, D J Beadle, L A King and I Bermudez

Journal of Receptors and Signal Transduction, Vol.15(1-4), pp.33-41

1995

DOI: https://doi.org/10.3109/10799899509045205

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Abstract

4 aminobutyric acid

4 aminobutyric acid A receptor

chloride channel

complementary DNA

neomycin

picrotoxin

receptor subunit

animal cell

Baculovirus

cell line

conference paper

controlled study

drug resistance

electrophysiology

gene expression

We have produced a stable insect cell line derived from Spodoptera frugiperda (Sf9) cells expressing a cDNA encoding a βsubunit of the Lymnaea stagnalis GABAA receptor. The cDNA was randomly integrated into the insect cell genome under the control of a baculovirus immediate early gene (IE-1) promoter. Stable cell lines were established by transformation of Sf9 cells with the expression vector pIEK1.LGβ1 together with a plasmid encoding a selectable marker which confers neomycin (G418) resistance. Following growth in the presence of G418, neomycin resistant clones were selected, amplified and analysed for the presence of functional GABA-gated chloride channels. Electrophysiological analysis of one cell line showed the presence of a picrotoxin-sensitive chloride channel not present in control Sf9 cells. These channels were also sensitive to GABA, albeit at relatively high (mM) concentrations. © 1995 Informa UK Ltd All rights reserved: reproduction in whole or part not permitted.

Details

Title: The production of a stably transformed insect cell line expressing an invertebrate GABAa receptor βsubunit
Authors: L A Smith (Author) - Oxford Brookes University, United Kingdom
M Amar (Author) - Oxford Brookes University, United Kingdom
Robert J Harvey (Author) - Universitat Hamburg, Germany
M G Darlison (Author) - Universitat Hamburg, Germany
F G P Earley (Author) - Jealott's Hill Research Station, United Kingdom
D J Beadle (Author) - Oxford Brookes University, United Kingdom
L A King (Author) - Oxford Brookes University, United Kingdom
I Bermudez (Author) - Oxford Brookes University, United Kingdom
Publication details: Journal of Receptors and Signal Transduction, Vol.15(1-4), pp.33-41
Publisher: Taylor & Francis Inc.
Date published: 1995
DOI: 10.3109/10799899509045205
ISSN: 1079-9893
Organisation Unit: School of Health; University of the Sunshine Coast, Queensland; School of Health and Sport Sciences - Legacy; Centre for Bioinnovation; School of Health and Behavioural Sciences - Legacy
Language: English
Record Identifier: 99450418902621
Output Type: Journal article

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