Description
Humana Press Inc. Pichia Protocols by James M. Cregg
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._x000D_ _x000D_
Table of Contents_x000D_
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Chapter 1 - Introduction [REVIEW CHAPTER]_x000D_
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Chapter 2 - Vectors and strains for expression [REVIEW CHAPTER]_x000D_
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Chapter 3 - DNA-Mediated Transformation_x000D_
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Chapter 4 - Rational design and optimization of fed-batch and continuous fermentations_x000D_
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Chapter 5 - Overexpression of a hookworm secretory protein saturates secretory capacity_x000D_
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Chapter 6 - Initial processing and purification strategies_x000D_
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Chapter 7 - Rapid screening of chromatography resins for the purification of proteins_x000D_
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Chapter 8 - Characterization of O-linked saccharides on glycoproteins_x000D_
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Chapter 9 - Modification of the N-glycosylation pathway to produce homogeneous human-like glycans using GlycoSwitch plasmids_x000D_
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Chapter 10 - N-linked glycan characterization of heterologous proteins_x000D_
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Chapter 11 - Heavy labeling of recombinant proteins_x000D_
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Chapter 12 - Selenomethionine labeling of recombinant proteins_x000D_
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Chapter 13 - Deuterium labeling of proteins_x000D_
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Chapter 14 - Classical genetic analysis_x000D_
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Chapter 15 - Identification of pexophagy genes by Restriction Enzyme-Mediated Integration (REMI)_x000D_
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Chapter 16 - Characterization of protein-protein interactions_x000D_
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Chapter 17 - Localization of proteins and organelles using fluorescence microscopy_x000D_
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Chapter 18 - Fluorescence microscopy and thin-section electron microscopy_x000D_