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Expression Screening of Integral Membrane Proteins by Fusion to Fluorescent Reporters

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The Next Generation in Membrane Protein Structure Determination

Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 922))

Abstract

The production of recombinant integral membrane proteins for structural and functional studies remains technically challenging due to their relatively low levels of expression. To address this problem, screening strategies have been developed to identify the optimal membrane sequence and expression host for protein production. A common approach is to genetically fuse the membrane protein to a fluorescent reporter, typically Green Fluorescent Protein (GFP) enabling expression levels, localization and detergent solubilisation to be assessed. Initially developed for screening the heterologous expression of bacterial membrane proteins in Escherichia coli, the method has been extended to eukaryotic hosts, including insect and mammalian cells. Overall, GFP-based expression screening has made a major impact on the number of membrane protein structures that have been determined in the last few years.

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Acknowledgments

The OPPF-UK is funded by the Medical Research Council, UK (grant MR/K018779/1).

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Authors and Affiliations

  1. OPPF-UK, The Research Complex at Harwell, Rutherford Appleton Laboratory Harwell, Oxford, UK

    Louise E. Bird, Joanne E. Nettleship, Valtteri Järvinen, Heather Rada, Anil Verma & Raymond J. Owens

  2. Division of Structural Biology, Henry Wellcome Building for Genomic Medicine, University of Oxford, Roosevelt Drive, Oxford, UK

    Louise E. Bird, Joanne E. Nettleship, Valtteri Järvinen, Heather Rada, Anil Verma & Raymond J. Owens

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  1. Louise E. Bird
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  2. Joanne E. Nettleship
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  3. Valtteri Järvinen
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  4. Heather Rada
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  5. Anil Verma
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  6. Raymond J. Owens
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Correspondence to Raymond J. Owens .

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Editors and Affiliations

  1. Membrane Protein Laboratory, Diamond Light Source/Imperial College London, Harwell Campus, Didcot, Oxfordshire, UK

    Isabel Moraes

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Bird, L.E., Nettleship, J.E., Järvinen, V., Rada, H., Verma, A., Owens, R.J. (2016). Expression Screening of Integral Membrane Proteins by Fusion to Fluorescent Reporters. In: Moraes, I. (eds) The Next Generation in Membrane Protein Structure Determination. Advances in Experimental Medicine and Biology, vol 922. Springer, Cham. https://doi.org/10.1007/978-3-319-35072-1_1

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