Pichia Protocols (eBook, PDF)

Redaktion: Cregg, James M.

• Format: PDF

Produktbeschreibung

The use of Pichia Pastoris as a recombinant protein system has increased greatly in popularity over the years. In Pichia Protocols 2^nd 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 2^nd Edition, is an essential tool for researchers working with the Pichia Pastoris system

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Produktdetails

• Verlag: Humana Press
• Seitenzahl: 268
• Erscheinungstermin: 3. Februar 2008
• Englisch
• ISBN-13: 9781597454568
• Artikelnr.: 64701668

Inhaltsangabe

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.

Rezensionen

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)