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Title High throughput protein expression and purification : methods and protocols / edited by Sharon A. Doyle
Published Totowa, N.J. : Humana Press ; [London] : [Springer, distributor], ©2009

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Description 1 online resource (xii, 322 pages) : illustrations (some color)
Series Methods in molecular biology ; 498
Methods in molecular biology (Clifton, N.J.) ; v. 498.
Contents High-throughput protein production (HTTP) : a review of enabling technologies to expedite protein production / Jim Koehn and Ian Hunt -- Designing experiments for high-throughput protein expression / Stephen P. Chambers and Susanne E. Swalley -- Gateway cloning for protein expression / Dominic Esposito, Leslie A. Garvey, and Chacko S. Chakiath -- Flexi vector cloning / Paul G. Blommel [and others] -- The precise engineering of expression vectors using high-throughput In-Fusion PCR cloning / Nick S. Berrow, David Alderton, and Raymond J. Owens -- The polymerase incomplete primer extension (PIPE) method applied to high-throughput cloning and site-directed mutagenesis / Heath E. Klock and Scott A. Lesley -- A family of LIC vectors for high-throughput cloning and purification of proteins / William H. Eschenfeldt [and others] -- "System 48" high-throughput cloning and protein expression analysis / James M. Abdullah, Andrzej Joachimiak, and Frank R. Collart -- Automated 96-well purification of hexahistidine-tagged recombinant proteins on MagneHis Ni²-particles / Chiann-Tso Lin, Priscilla A. Moore, and Vladimir Kery -- E. coli and insect cell expression, automated purification and quantitative analysis / StephenP. Chambers [and others] -- Hexahistidine-tagged maltose-binding protein as a fusion partner for the production of soluble recombinant proteins in Escherichia coli / Brian P. Austin, Sreedevi Nallamsetty, and David S. Waugh -- PHB-intein-mediated protein purification strategy / Alison R. Gillies, Mahmoud Reza Banki, and David W. Wood -- High-throughput biotinylation of proteins / Brian K. Kay, Sang Thai, and Veronica V. Volgina -- High-throughput insect cell protein expression applications / Mirjam Buchs [and others] -- High-throughput protein expression using cell-free system / Kalavathy Sitaraman and Deb K. Chatterjee -- The production of glycoproteins by transient expression in mammalian cells / Joanne E. Nettleship, Nahid Rahman-Huq, and Raymond J. Owens -- High-throughput expression and detergent screening of integral membrane proteins / Said Eshaghi -- Cell-free expression for nanolipoprotein particles : building a high-throughput membrane protein solubility platform / Jenny A. Cappuccio [and others] -- Expression and purification of soluble His₆-tagged TEV protease / Joseph E. Tropea, Scott Cherry, and David S. Waugh -- High-throughput protein concentration and buffer exchange : comparison of ultrafiltration and ammonium sulfate precipitation / Priscilla A. Moore and Vladimir Kery
Summary Despite exciting advances in genome sequencing, isolating a protein from its expression system in its native form still presents a complex challenge. In High Throughput Protein Expression and Purification: Methods and Protocols, leading scientists detail the most successful protocols currently in use, including various high throughput cloning schemes, protein expression analysis, and production protocols. This volume describes the use of E. coli, insect, and mammalian cells, as well as cell-free systems for the production of a wide variety of proteins, including glycoproteins and membrane proteins, in order to best represent strategies that create and exploit common features to enable simplified cloning, stable expression, and purification of proteins. Written in the highly successful Methods in Molecular Biology series format, the chapters present brief introductions to the subject, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and a Notes section for tips on troubleshooting and avoiding known pitfalls. Cutting-edge and comprehensive, High Throughput Protein Expression and Purification: Methods and Protocols is an ideal reference for protein biochemists and all those who wish to apply these easy-to-use protocols to the many applicable fields
Bibliography Includes bibliographical references and index
Notes English
Print version record
Subject Proteins -- Separation -- Methodology
Gene expression.
Gene Expression
Proteins -- genetics
Proteins -- isolation & purification
Proteomics -- methods
Gene expression
Animal Biochemistry.
Biophysics.
Human Anatomy & Physiology.
Biology.
Health & Biological Sciences.
Form Electronic book
Author Doyle, Sharon A. (Sharon Ann)
ISBN 9781597451963
1597451967