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by Roslyn M. Bill (Editor)
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
Back Jacket
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.
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