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Pichia Protocols: Methods in Molecular Biology, cartea 389

Editat de James M. Cregg
en Limba Engleză Paperback – 19 noi 2010
Pichia Protocols 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).
This volume fully updates and expands upon the first edition of Pichia Protocols, and focuses primarily on information that has come to light since its original publication. Each chapter presents cutting-edge and cornerstone protocols for utilizing P. pastoris as a model recomibinant protein production system.
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Specificații

ISBN-13: 9781617375637
ISBN-10: 1617375632
Pagini: 268
Ilustrații: XII, 268 p.
Dimensiuni: 152 x 229 x 16 mm
Greutate: 0.42 kg
Ediția:Softcover reprint of hardcover 2nd ed. 2007
Editura: Humana Press Inc.
Colecția Humana
Seria Methods in Molecular Biology

Locul publicării:Totowa, NJ, United States

Public țintă

Research

Cuprins

Vectors and Strains for Expression.- DNA-Mediated Transformation.- Rational Design and Optimization of Fed-Batch and Continuous Fermentations.- Saturation of the Secretory Pathway by Overexpression of a Hookworm (Necator americanus) Protein (Na-ASP1).- Purification of the N- and C-Terminal Subdomains of Recombinant Heavy Chain Fragment C of Botulinum Neurotoxin Serotype C.- Rapid Screening of Chromatography Resins for the Purification of Proteins.- Characterization of O-Linked Saccharides on Glycoproteins.- Modification of the N-Glycosylation Pathway to Produce Homogeneous, Human-Like Glycans Using GlycoSwitch Plasmids.- N-Linked Glycan Characterization of Heterologous Proteins.- Heavy Labeling of Recombinant Proteins.- Selenomethionine Labeling of Recombinant Proteins.- Selective Isotopic Labeling of Recombinant Proteins Using Amino Acid Auxotroph Strains.- Classical Genetics.- Identification of Pexophagy Genes by Restriction Enzyme-Mediated Integration.- Characterization of Protein-Protein Interactions.- Localization of Proteins and Organelles Using Fluorescence Microscopy.- Fluorescence Microscopy and Thin-Section Electron Microscopy.

Recenzii

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)

Textul de pe ultima copertă

The use of Pichia Pastoris as a recombinant protein system has increased greatly in popularity over the years. In Pichia Protocols 2nd Edition, the author focuses on new information that has come to light in recent years but repeats key basic methods so that researchers are not absolutely dependent on having both editions in order to utilize the system. The chapters in this indispensable laboratory aid cover a wide range of topics associated with Pichia Pastoris. The use of fermentors to grow Pichia Pastoris and good advice on how to go about the initial processing of material from these cultures is discussed. An extensive discussion on how to formally analyze mutations in essential P. pastoris genes is also presented. The final chapters are for cell biologists who are for cell biologists using the Pichia Pastoris system to investigate protein interactions, or to localize proteins in yeast cells. Pichia Protocols 2nd Edition, is an essential tool for researchers working with the Pichia Pastoris system

Caracteristici

All protocols are tested and readily and reproducible Updates and expands upon the first volume Presents the basics of Pichia pastoris and describes available vectors and host strains for expression