Recombinant protein production in yeast : methods and protocols

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書誌事項

Recombinant protein production in yeast : methods and protocols

edited by Roslyn M. Bill

（Methods in molecular biology / John M. Walker, series editor, 866）（Springer protocols）

Humana Press, c2012

  • : pbk

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注記

Includes bibliographical references and index

内容説明・目次

内容説明

In the last few years, significant advances have been made in understanding how a yeast cell responds to the stress of producing a recombinant protein, and how this information can be used to engineer improved host strains. The molecular biology of the expression vector, through the choice of promoter, tag and codon optimization of the target gene, is also a key determinant of a high-yielding protein production experiment. Recombinant Protein Production in Yeast: Methods and Protocols examines the process of preparation of expression vectors, transformation to generate high-yielding clones, optimization of experimental conditions to maximize yields, scale-up to bioreactor formats and disruption of yeast cells to enable the isolation of the recombinant protein prior to purification. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Recombinant Protein Production in Yeast: Methods and Protocols, seeks to aid scientists in adopting yeast as a protein production host.

目次

1 Optimising Yeast as a Host for Recombinant Protein Production( review) Nicklas Bonander and Roslyn M Bill 2 Which Yeast Species Shall I Choose? Saccharomyces cerevisiae versus Pichia pastoris (review) Richard AJ Darby, Stephanie P Cartwright, Marvin V. Dilworth and Roslyn M Bill 3 Preparation of Pichia pastoris Expression Plasmids Christel Logez, Fatima Alkhalfioui, Bernadette Byrne, Renaud Wagner 4 Preparation of Saccharomyces cerevisiae Expression Plasmids David Drew and Hyun Kim 5 Codon Optimisation for Heterologous Gene Expression in Yeast Kristina Hedfalk 6 Yeast Transformation to Generate High-yielding Clones Mohammed Jamshad and Richard AJ Darby 7 Screening for High-yielding Pichia pastoris Clones: the Production of G Protein-Coupled Receptors as a Case Study Shweta Singh, Adrien Gras, Cedric Fiez-Vandal, Magdalena Martinez, Renaud Wagner, Bernadette Byrne 8 Screening for High-yielding Saccharomyces cerevisiae Clones: Using a Green Fluorescent Protein Fusion Strategy in the Production of Membrane Proteins David Drew and Hyun Kim 9 The Effect of Antifoam Addition on Protein Production Yields Sarah J Routledge and Roslyn M Bill 10 Setting up a Bioreactor for Recombinant Protein Production in Yeast Sarah J Routledge and Michelle Clare 11 The Implementation of a Design of Experiments Strategy to Increase Recombinant Protein Yields in Yeast (review) Nagamani Bora, Zharain Bawa, Roslyn M. Bill and Martin D. B. Wilks 12 On-line Analysis and Process Control in Recombinant Protein Production (review) Shane M. Palmer and Edmund R.S. Kunji 13 Monitoring the Biomass Accumulation of Recombinant Yeast Cultures: Off-line Estimations of Dry Cell Mass and Cell Counts Shane Palmer and Edmund R S Kunji 14 On-line Monitoring of Biomass Accumulation in Recombinant Yeast Cultures Shane Palmer and Edmund R S Kunji 15 Optimising P. pastoris Induction Zharain Bawa and Richard AJ Darby 16 Optimizing S. cerevisiae Induction Regimes David Drew and Hyun Kim 17 Large Scale Production of Membrane Proteins in Pichia pastoris: The Production of G Protein-coupled Receptors as a Case Study Shweta Singh, Adrien Gras, Cedric Fiez-Vandal, Magdalena Martinez, Renaud Wagner and Bernadette Byrne 18 Large-scale Production of Membrane Proteins in Saccharomyces cerevisiae: Using a Green Fluorescent Protein Fusion Strategy in the Production of Membrane Proteins David Drew and Hyun Kim 19 Large Scale Production of Secreted Proteins in Pichia pastoris Nagamani Bora 20 Disruption of Yeast Cells to Isolate Recombinant Proteins Mohammed Jamshad and Richard AJ Darby

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