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

L. A. Smith School of Biological and Molecular Sciences, Oxford Brookes University, Oxford, UK

M. Amar School of Biological and Molecular Sciences, Oxford Brookes University, Oxford, UK

R. J. Harvey Institüt fur Zelibiochemie und Klinische Neurobiologie, Universität Hamburg, Hamburg, Germany

M. G. Darlison Institüt fur Zelibiochemie und Klinische Neurobiologie, Universität Hamburg, Hamburg, Germany

F. G. P. Earley Zeneca Agrochemicals, Jealotts Hill Research Station, Bracknell, UK

D. J. Beadle School of Biological and Molecular Sciences, Oxford Brookes University, Oxford, UK

L. A. King School of Biological and Molecular Sciences, Oxford Brookes University, Oxford, UK

I. Bermudez School of Biological and Molecular Sciences, Oxford Brookes University, Oxford, UK

show all

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 GABA_A 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.

Reprints and Corporate Permissions

Please note: Selecting permissions does not provide access to the full text of the article, please see our help page How do I view content?

To request a reprint or corporate permissions for this article, please click on the relevant link below:

Order Reprints Request Corporate Permissions

Academic Permissions

Please note: Selecting permissions does not provide access to the full text of the article, please see our help page How do I view content?

Obtain permissions instantly via Rightslink by clicking on the button below:

Request Academic Permissions

If you are unable to obtain permissions via Rightslink, please complete and submit this Permissions form. For more information, please visit our Permissions help page.

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

Information for

Open access

Opportunities

Help and information

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

Abstract

Reprints and Corporate Permissions

Academic Permissions

Related research

To cite this article:

Download citation

Information for

Open access

Opportunities

Help and information

Keep up to date

Your download is now in progress and you may close this window

Login or register to access this feature

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