title: Screening for high-yielding Pichia pastoris clones: the production of G protein-coupled receptors as a case study.
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.
authors.affiliation: Division of Molecular Biosciences, Imperial College London, London, UK.
authors.forename: Shweta
authors.initials: S
authors.lastname: Singh
authors.forename: Adrien
authors.initials: A
authors.lastname: Gras
authors.forename: Cedric
authors.initials: C
authors.lastname: Fiez-Vandal
authors.forename: Magdalena
authors.initials: M
authors.lastname: Martinez
authors.forename: Renaud
authors.initials: R
authors.lastname: Wagner
authors.forename: Bernadette
authors.initials: B
authors.lastname: Byrne
mesh_terms: D015153:Blotting, Western; D003001:Cloning, Molecular / Q000379:methods; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D043562:Receptors, G-Protein-Coupled; D011994:Recombinant Proteins