Pichia protocols
著者
書誌事項
Pichia protocols
（Methods in molecular biology / John M. Walker, series editor, v. 103）
Humana Press, c1998
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注記
Includes bibliographical references and index
内容説明・目次
内容説明
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.
目次
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 a.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.
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