Abstract
Pichia pastoris is an established host for the production of a wide range of recombinant proteins including membrane proteins. The system has a particularly good track record for the production of G protein-coupled receptors (GPCRs). Generation and screening of expression clones with this system use standard molecular biology techniques. Multiple clones can be generated and screened in a matter of a few weeks making this similar to Escherichia coli in terms of speed. In addition, basic buffer components and the lack of expensive equipment make small-scale expression screening in P. pastoris very cost-effective. Here we describe the procedures used for small-scale GPCR production screening.
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This research was funded by the MepNet consortium, BBSRC, and GlaxoSmithKline.
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Singh, S., Gras, A., Fiez-Vandal, C., Martinez, M., Wagner, R., Byrne, B. (2012). Screening for High-Yielding Pichia pastoris Clones: The Production of G Protein-Coupled Receptors as a Case Study. 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_7
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DOI: https://doi.org/10.1007/978-1-61779-770-5_7
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