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Pichia Protocols

  • Book
  • Apr 2014

Overview

Editors:
  1. David R. Higgins

    1. Chiron Technologies/Center for Gene Therapy, San Diego, USA

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    You can also search for this editor in PubMed   Google Scholar

  2. James M. Cregg

    1. Oregon Graduate Institute of Science and Technology, Portland, USA

    View editor publications

    You can also search for this editor in PubMed   Google Scholar

Part of the book series: Methods in Molecular Biology (MIMB, volume 103)

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Table of contents (17 protocols)

  1. Front Matter

    Pages i-xi
    Download chapter PDF

  2. Introduction to Pichia pastoris

    • David R. Higgins, James M. Cregg
    Pages 1-15

  3. Classical Genetic Manipulation

    • James M. Cregg, Shigang Shen, Monique Johnson, Hans R. Waterham
    Pages 17-26

  4. Transformation

    • James M. Cregg, Kimberly A. Russell
    Pages 27-39

  5. Small Vectors for Expression Based on Dominant Drug Resistance with Direct Multicopy Selection

    • David R. Higgins, Katie Busser, John Comiskey, Peter S. Whittier, Thomas J. Purcell, James P. Hoeffler
    Pages 41-53

  6. The Generation of Multicopy Recombinant Strains

    • Mike Romanos, Carol Scorer, Koti Sreekrishna, Jeff Clare
    Pages 55-72

  7. Isolation of Nucleic Acids

    • David J. Miles, Katie Busser, Christine Stalder, David R. Higgins
    Pages 73-80

  8. Generation of Protease-Deficient Strains and Their Use in Heterologous Protein Expression

    • Martin A. G. Gleeson, Christopher E. White, David P. Meininger, Elizabeth A. Komives
    Pages 81-94

  9. Glycosylation Profiling of Heterologous Proteins

    • José A. Cremata, Raquel Montesino, Omar Quintero, Rossana García
    Pages 95-105

  10. High Cell-Density Fermentation

    • Jayne Stratton, Vijay Chiruvolu, Michael Meagher
    Pages 107-120

  11. Use of Pichia pastoris as a Model Eukaryotic System

    • Klaas Nico Faber, Ype Elgersma, John A. Heyman, Antonius Koller, Georg H. Lüers, William M. Nuttley et al.
    Pages 121-147

  12. Secretion of Recombinant Human Insulin-Like Growth Factor I (IGF-I)

    • Russell A. Brierley
    Pages 149-177

  13. Secretion of scFv Antibody Fragments

    • Hermann Gram, Rita Schmitz, Rüdiger Ridder
    Pages 179-192

  14. Expression of Tetanus Toxin Fragment C

    • Jeff Clare, Koti Sreekrishna, Mike Romanos
    Pages 193-208

  15. Expression of EGF and HIV Envelope Glycoprotein

    • Jeff Clare, Carol Scorer, Rich Buckholz, Mike Romanos
    Pages 209-225

  16. Expression of an Integral Membrane Protein, the 5HT5A Receptor

    • H. Markus Weiß, Winfried Haase, Helmut Reiländer
    Pages 227-239

  17. Localization of the myc-Tagged 5HT5A Receptor by Immunogold Staining of Ultrathin Sections

    • Winfried Haase, H. Markus Weiß, Helmut Reiländer
    Pages 241-247

  18. Appendix

    • Winfried Haase, H. Markus Weiß, Helmut Reiländer
    Pages 249-261

  19. Back Matter

    Pages 263-270
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Keywords

About this book

David Higgins and James Cregg compile for the first time an all-inclusive collection of key experimental procedures to manipulate Pichia pastoris for gene expression purposes. The methods-developed by world-class researchers in their own laboratories-contain detailed descriptions of available expression strains, as well as vectors and step-by-step instructions for the construction of expression strains and for their use in the production of foreign proteins from shake-flask or fermentor cultures. General procedures for the purification of foreign proteins are also provided, along with protocols for analyzing foreign protein products for glycosylation and subcellular location. Its detailed instructions will permit any researcher in molecular biology to quickly construct an expression strain that will produce a desired foreign protein, and to use this state-of-the-art molecular and cellular biological system successfully in every area of biological research today.

Reviews

"In this valuable Methods in Molecular Biology volume, the editors have assembled an international cast of Pichia experts who provide detailed protocols, advice, and theory on recombinant expression in this methylotrophic yeast. . .a focused writing style that is enjoyable to read. Molecular geneticists are afforded the essential information to achieve success in vector construction, transformation and screening."-The Quarterly Review of Biology

Editors and Affiliations

  • Chiron Technologies/Center for Gene Therapy, San Diego, USA

    David R. Higgins

  • Oregon Graduate Institute of Science and Technology, Portland, USA

    James M. Cregg

Bibliographic Information

  • Book Title: Pichia Protocols

  • Editors: David R. Higgins, James M. Cregg

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1385/0896034216

  • Publisher: Humana Totowa, NJ

  • eBook Packages: Springer Protocols

  • Copyright Information: Humana Press 1998

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XI, 270

  • Topics: Cell Biology

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