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Pichia Protocols James M. Cregg

Pichia Protocols By James M. Cregg

Pichia Protocols by James M. Cregg


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Summary

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).

Pichia Protocols Summary

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.

Pichia Protocols Reviews

From the reviews of the second edition:

P. pastoris has many superior traits. ... can grow to very high cell densities prior to switching on an inducible promoter (the AOX1 gene) for maximal recombinant protein production. The diversity of such proteins produced by expression strains of P. pastoris is quite amazing and this book describes detailed experimental protocols for a few of them (e.g. hookworm protein, botulinum toxin). ... This is a stand-alone book ... . a book that will prove valuable to yeast molecular geneticists and biotechnologists. (Graeme Walker, Microbiology Today, May, 2008)

Table of Contents

Table of Contents Chapter 1 - Introduction [REVIEW CHAPTER] Chapter 2 - Vectors and strains for expression [REVIEW CHAPTER] Chapter 3 - DNA-Mediated Transformation Chapter 4 - Rational design and optimization of fed-batch 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 O-linked saccharides on glycoproteins Chapter 9 - Modification of the N-glycosylation pathway to produce homogeneous human-like glycans using GlycoSwitch plasmids Chapter 10 - N-linked 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 Enzyme-Mediated Integration (REMI) Chapter 16 - Characterization of protein-protein interactions Chapter 17 - Localization of proteins and organelles using fluorescence microscopy Chapter 18 - Fluorescence microscopy and thin-section electron microscopy

Additional information

NLS9781617375637
9781617375637
1617375632
Pichia Protocols by James M. Cregg
New
Paperback
Humana Press Inc.
2010-11-19
268
N/A
Book picture is for illustrative purposes only, actual binding, cover or edition may vary.
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Customer Reviews - Pichia Protocols