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  Semliki Forest Virus vectors for overexpression of 101 G Protein-coupled receptors in mammalian host cells

Hassaine, G., Wagner, R., Kempf, J., Cherouati, N., Hassaine, N., Prual, C., et al. (2006). Semliki Forest Virus vectors for overexpression of 101 G Protein-coupled receptors in mammalian host cells. Protein Expression and Purification, 45(2), 343-351. doi:10.1016/j.pep.2005.06.007.

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 Creators:
Hassaine, Gherici1, Author
Wagner, Renaud2, Author
Kempf, Juliette2, Author
Cherouati, Nadja2, Author
Hassaine, Nhouzha1, Author
Prual, Cécile3, Author              
André, Nicolas3, Author              
Reinhart, Christoph3, Author              
Pattus, Franc2, Author
Lundstrom, Kenneth1, Author
Affiliations:
1BioXtal, Chemin des Croisettes 22, CH-1066 Epalinges, Switzerland, ou_persistent22              
2Louis Pasteur University, Illkirch, France, ou_persistent22              
3Department of Molecular Membrane Biology, Max Planck Institute of Biophysics, Max Planck Society, ou_2068290              

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Free keywords: G protein-coupled receptors; Semliki Forest virus; Structural genomics Structural biology; Recombinant expression; Large-scale production; Mammalian cell lines; Immunodetection; Binding assays
 Abstract: 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.

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Language(s): eng - English
 Dates: 2005-06-062005-05-142005-07-112006-02
 Publication Status: Published in print
 Pages: 9
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1016/j.pep.2005.06.007
 Degree: -

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Title: Protein Expression and Purification
Source Genre: Journal
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Publ. Info: San Diego : Academic Press
Pages: - Volume / Issue: 45 (2) Sequence Number: - Start / End Page: 343 - 351 Identifier: ISSN: 1046-5928
CoNE: https://pure.mpg.de/cone/journals/resource/954922650158