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pubmed-article:9234898pubmed:abstractTextG-protein-coupled or 7-transmembrane receptors (7TMRs) are often studied after heterologous expression in mammalian cells such as COS-7, CHO-K1, or HEK-293s. In this paper, we describe the development of a rapid and generic method for producing stable Chinese hamster ovary cell lines expressing high levels of recombinant 7TMRs by N-terminal tagging these proteins with the hemagglutinin (HA) sequence. To illustrate the broad applicability of this technique, we have presented data from cell lines expressing a glycoprotein hormone receptor for follicle-stimulating hormone (FSHR), CXC- (CXCR-2), and CC-chemokine (CCR-1) receptors and peptide receptors from the somatostatin (SSTR1, 2, 5) and neuropeptide Y (NPY-Y2, -Y4 Rs) families. Typically, cell lines with a receptor density of 1 to 15 pmol/mg protein are produced with this method. The presence of the HA tag does not adversely affect the binding or functional activity of the receptors.lld:pubmed
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pubmed-article:9234898pubmed:articleTitleA generic method for the production of cell lines expressing high levels of 7-transmembrane receptors.lld:pubmed
pubmed-article:9234898pubmed:affiliationAffymax Research Institute, Palo Alto, California 94304, USA. Kerry_Koller@qmgates.affymax.comlld:pubmed
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