Abstract
The production of membrane proteins in the large quantities necessary for structural analysis requires many optimization steps. The GFP-fusion-based scheme described in earlier chapters (Chapters 4, 8, and 16) facilitates these steps by allowing the selection of high yielding clones that produce detergent-stable membrane proteins. Here, we describe the experimental steps required to establish the reproducible, large-scale production and purification of membrane protein–GFP fusions using S. cerevisiae.
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Acknowledgments
This work was supported by the Royal Society (United Kingdom) through a University Research Fellowship to DD and by a Basic Science Research Program grant through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (NRF0409-20100093) to HK.
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Drew, D., Kim, H. (2012). Large-Scale Production of Membrane Proteins in Saccharomyces cerevisiae: Using a Green Fluorescent Protein Fusion Strategy in the Production of Membrane Proteins. In: Bill, R. (eds) Recombinant Protein Production in Yeast. Methods in Molecular Biology, vol 866. Humana Press. https://doi.org/10.1007/978-1-61779-770-5_18
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DOI: https://doi.org/10.1007/978-1-61779-770-5_18
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