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Semliki Forest virus vectors for overexpression of 101 G protein-coupled receptors in mammalian host cells.
BioXtal, Chemin des Croisettes 22, CH-1066 Epalinges, Switzerland.
Semliki Forest virus vectors were applied for the evaluation of 101 G protein-coupled receptors in three mammalian cell lines. Western blotting demonstrated that 95 of the 101 tested GPCRs showed positive signals. A large number of the GPCRs were expressed at high levels suggesting receptor yields in the range of 1 mg/L or higher, suitable for structural biology applications. Specific binding assays on a selected number of GPCRs were carried out to compare the correlation between total and functional protein expression. Ligands and additives supplemented to the cell culture medium were evaluated for expression enhancement. Selected GPCRs were also expressed from mutant SFV vectors providing enhanced protein expression and reduced host cell toxicity in attempts to further improve receptor yields.
PMID: 16055346 [PubMed - indexed for MEDLINE]
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Cited by 2 PubMed Central articles
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Using EGFP fusions to monitor the functional expression of GPCRs in the Drosophila Schneider 2 cells.
Brillet K, Perret BG, Klein V, Pattus F, Wagner R.
Cytotechnology. 2008 May; 57(1):101-9. Epub 2008 Jan 19.
[Cytotechnology. 2008]
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Large-scale functional expression of WT and truncated human adenosine A2A receptor in Pichia pastoris bioreactor cultures.
Singh S, Gras A, Fiez-Vandal C, Ruprecht J, Rana R, Martinez M, Strange PG, Wagner R, Byrne B.
Microb Cell Fact. 2008 Oct 10; 7:28. Epub 2008 Oct 10.
[Microb Cell Fact. 2008]