Please use this identifier to cite or link to this item: http://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), Ch.8, pp.115-129
Publisher: Humana Press
Issue Date: 2009
Series/Report no.: Methods in Molecular Biology; 552
ISBN: 9781603273176
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
RMID: 0030012992
DOI: 10.1007/978-1-60327-317-6_8
Appears in Collections:Molecular and Biomedical Science publications

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