High throughput protein expression and purification: methods and protocols

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 ofandnbsp;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. Provides step-by-step protocols for protein production that utilize a wide range of cutting-edge techniques Covers cell-based and cell-free methodologies neededto produce bacterial proteins, eukaryotic proteins, soluble and membrane proteins Examines a variety of cloning methodologies that enable high-throughput cloning for protein production INDICE: From the contents High Throughput Protein Production (HTPP): A Review of Enabling Technologies to Expedite Protein Production.- Designing Experiments for High-Throughput Protein Expression.- Gateway Cloning for Protein Expression.- Flexi Vector Cloning.- The Precise Engineering of Expression VectorsUsing High Throughput In-Fusion™ PCR Cloning.- The Polymerase Incomplete Primer Extension (PIPE) Method Applied to High-Throughput Cloning and Site-Directed Mutagenesis.- A Family of LIC Vectors for High-Throughput Cloning and Purification of Proteins.- "System 48" High Throughput Cloning and Protein Expression Analysis.- Automated 96-Well Purification of Hexahistidine-Tagged Recombinant Proteins on MagneHis Ni2+-Particles.- E.coli and Insect Cell Expression, Automated Purification and Quantitative Analysis.- Hexahistidine-Tagged Maltose-Binding Protein as a Fusion Partner for the Production of Soluble Recombinant Proteins in Escherichia coli.- PHB-Intein-Mediated Protein Purification Strategy.

• ISBN: 978-1-58829-879-9
• Editorial: Humana
• Encuadernacion: Cartoné
• Páginas: 305
• Fecha Publicación: 01/11/2008
• Nº Volúmenes: 1
• Idioma: Inglés