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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.
Introduction to Pichia pastoris David R. Higgins and James M. Cregg. Classical Genetic Manipulation James M. Cregg Shigang Shen Monique Johnson and Hans R. Waterham. Transformation James M. Cregg and Kimberly A. Russell. 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 and James P. Hoeffler. The Generation of Multicopy Recombinant Strains Mike Romanos Carol Scorer Koti Sreekrishna and Jeff Clare. Isolation of Nucleic Acids David J. Miles Katie Busser Christine Stalder and David R. Higgins. Generation of Protease- Deficient Strains and Their Use in Heterologous Protein Expression Martin A. G. Gleeson Christopher E. White David P. Meininger and Elizabeth A. Komives. Glycosylation Profiling of Heterologous Proteins Jos' Cremata Raquel Montesino Omar Quintero and Rossana Garc├┬├┬ía.High Cell-Density Fermentation Jayne Stratton Vijay Chiruvolu and Michael Meagher. Use of Pichia pastoris as a Model Eukaryotic System: Peroxisome Biogenesis Klaas Nico Faber Ype Elgersma John A. Heyman Antonius Koller Georg H. L?ers William M. Nuttley Stanley R. Terlecky Thibaut J. Wenzel and Suresh Subramani. Secretion of Recombinant Human Insulin-Like Growth Factor I (IGF-I) Russell A. Brierley. Secretion of scFv Antibody Fragments Hermann Gram Rita Schmitz and R?diger Ridder. Expression of Tetanus Toxin Fragment C Jeff Clare Koti Sreekrishna and Mike Romanos. Expression of EGF and HIV Envelope Glycoprotein Jeff Clare Carol Scorer Rich Buckholz and Mike Romanos. Expression of an Integral Membrane Protein the 5HT5A Receptor H. Markus Weia Winfried Haase and Helmut Reil"nder. Localization of the myc-Tagged 5HT5A Receptor by Immunogold Staining of Ultrathin Sections Winfried Haase H. Markus Weia and Helmut Reil"nder. Appendix: Foreign Proteins Expressed in P. pastoris.Index.
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