Screening for High-Yielding Pichia pastoris Clones: The Production of G Protein-Coupled Receptors as a Case Study
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.
Key wordsSmall-scale expression Western blot analysis Functional analysis Optimization trials
This research was funded by the MepNet consortium, BBSRC, and GlaxoSmithKline.
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