The Production of a Stably Transformed Insect Cell Line Expressing An Invertebrate GABA A Receptor β-Subunit

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

Abstract

We have produced a stable insect cell line derived from Spodoptera frugiperda (Sf9) cells expressing a cDNA encoding a β-subunit 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.

Citation

Smith, L. A., Amar, M., Harvey, R. J., Darlison, M. G., Earley, F. G. P., Beadle, D. J., King, L. A., & Bermudez, I. (1995). The Production of a Stably Transformed Insect Cell Line Expressing An Invertebrate GABA A Receptor β-Subunit. Journal of Receptors and Signal Transduction, 15(1-4), 33-41. https://doi.org/10.3109/10799899509045205

• Journal Article Type: Article
• Acceptance Date: Jan 1, 1995
• Online Publication Date: Sep 26, 2008
• Publication Date: 1995-01
• Deposit Date: Aug 1, 2016
• Journal: Journal of Receptors and Signal Transduction
• Print ISSN: 1079-9893
• Electronic ISSN: 1532-4281
• Publisher: Taylor & Francis
• Peer Reviewed: Peer Reviewed
• Volume: 15
• Issue: 1-4
• Pages: 33-41
• DOI: https://doi.org/10.3109/10799899509045205
• Keywords: Spodoptera frugiperda (Sf9), Stable cell lines, plasmid encoding, Electrophysiological analysis,
• Public URL: http://researchrepository.napier.ac.uk/Output/321025