Expression of Eukaryotic Membrane Proteins in Pichia pastoris.
Fiche publication
Date publication
janvier 2016
Journal
Methods in molecular biology (Clifton, N.J.)
Auteurs
Membres identifiés du Cancéropôle Est :
Dr WAGNER Renaud
Tous les auteurs :
Hartmann L, Kugler V, Wagner R
Lien Pubmed
Résumé
A key point when it comes to heterologous expression of eukaryotic membrane proteins (EMPs) is the choice of the best-suited expression platform. The yeast Pichia pastoris has proven to be a very versatile system showing promising results in a growing number of cases. Indeed, its particular methylotrophic characteristics combined to the very simple handling of a eukaryotic microorganism that possesses the majority of mammalian-like machineries make it a very competitive expression system for various complex proteins, in amounts compatible with functional and structural studies. This chapter describes a set of robust methodologies routinely used for the successful expression of a variety of EMPs, going from yeast transformation with the recombinant plasmid to the analysis of the quality and quantity of the proteins produced.
Mots clés
Cloning, Molecular, Eukaryotic Cells, metabolism, Membrane Proteins, genetics, Pichia, genetics, Plasmids, genetics, Protein Engineering, Recombinant Proteins, metabolism, Transformation, Genetic
Référence
Methods Mol. Biol.. 2016 ;1432:143-62