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.
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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
The use of Pichia Pastoris as a recombinant protein system has increased greatly in popularity over the years. In Pichia Protocols 2nd Edition, the author focuses on new information that has come to light in recent years but repeats key basic methods so that researchers are not absolutely dependent on having both editions in order to utilize the system. The chapters in this indispensable laboratory aid cover a wide range of topics associated with Pichia Pastoris. The use of fermentors to grow Pichia Pastoris and good advice on how to go about the initial processing of material from these cultures is discussed. An extensive discussion on how to formally analyze mutations in essential P. pastoris genes is also presented. The final chapters are for cell biologists who are for cell biologists using the Pichia Pastoris system to investigate protein interactions, or to localize proteins in yeast cells. Pichia Protocols 2nd Edition, is an essential tool for researchers working with the Pichia Pastoris system
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