Beschreibung
This book focuses on recent developments of Pichia pastoris as a recombinant protein production system. Highlighted topics include a discussion on the use of fermentors to grow Pichia pastoris, information on the O- and N-linked glycosylation, methods for labeling Pichia pastoris expressed proteins for structural studies, and the introduction of mutations in Pichia pastoris genes by the methods of restriction enzyme-mediated integration (REMI). Each chapter presents cutting-edge and cornerstone protocols for utilizing P. pastoris as a model recomibinant protein production system. This volume fully updates and expands upon the first edition.
Autorenporträt
InhaltsangabeTable of Contents Chapter 1 Introduction [REVIEW CHAPTER] Chapter 2 Vectors and strains for expression [REVIEW CHAPTER] Chapter 3 DNAMediated Transformation Chapter 4 Rational design and optimization of fedbatch and continuous fermentations Chapter 5 Overexpression of a hookworm secretory protein saturates secretory capacity Chapter 6 Initial processing and purification strategies Chapter 7 Rapid screening of chromatography resins for the purification of proteins Chapter 8 Characterization of Olinked saccharides on glycoproteins Chapter 9 Modification of the Nglycosylation pathway to produce homogeneous humanlike glycans using GlycoSwitch plasmids Chapter 10 Nlinked glycan characterization of heterologous proteins Chapter 11 Heavy labeling of recombinant proteins Chapter 12 Selenomethionine labeling of recombinant proteins Chapter 13 Deuterium labeling of proteins Chapter 14 Classical genetic analysis Chapter 15 Identification of pexophagy genes by Restriction EnzymeMediated Integration (REMI) Chapter 16 Characterization of proteinprotein interactions Chapter 17 Localization of proteins and organelles using fluorescence microscopy Chapter 18 Fluorescence microscopy and thinsection electron microscopy
