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https://hdl.handle.net/2440/87599
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Type: | Book chapter |
Title: | GPCR expression using baculovirus-infected Sf9 cells |
Author: | Aloia, A.L. Glatz, R.V. McMurchie, E.J. Leifert, W.R. |
Citation: | G Protein-Coupled Receptors in Drug Discovery, 2009 / Leifert, W.R. (ed./s), vol.552, Ch.8, pp.115-129 |
Publisher: | Humana Press |
Issue Date: | 2009 |
Series/Report no.: | Methods in Molecular Biology; 552 |
ISBN: | 9781603273176 |
Editor: | Leifert, W.R. |
Statement of Responsibility: | Amanda L. Aloia, Richard V. Glatz, Edward J. McMurchie, and Wayne R. Leifert |
Abstract: | Expression of proteins in insect cells using recombinant baculoviruses has gained wide use in the G protein-coupled receptor (GPCR) community. This expression system produces high yields of functional receptor, is able to perform post-translational modifications, and is readily adaptable to large-scale culture. Here, we describe the generic methods for expressing a GPCR using baculovirus-infected insect cells, including the maintenance of insect cell culture. Data are presented for polyhedrin promoter-driven expression of a C-terminal 6 x histidine-tagged mammalian M(2) muscarinic receptor in Sf9 cells. Results demonstrate that expressed receptor could be detected and quantified using radiolabeled ligand binding, that expression was maximal at approximately 72 h post-infection, and that expression levels could be altered by addition of various ligands to cultures of infected insect cells. |
Keywords: | Baculovirus; Insect cell; GPCR expression; M2 muscarinic receptor |
Rights: | © Humana Press, a part of Springer Science+Business Media, LLC 2009 |
DOI: | 10.1007/978-1-60327-317-6_8 |
Published version: | http://dx.doi.org/10.1007/978-1-60327-317-6_8 |
Appears in Collections: | Aurora harvest 2 Molecular and Biomedical Science publications |
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