E. Coli Gene Expression Protocols

ISBN-13: 9781617373022 / Angielski / Miękka / 2011 / 347 str.

Peter E. Vaillancourt

Opis

Peter E. Vaillancourt presents a collection of popular and emerging methodologies that take advantage of E. coli's ability to quickly and inexpensively express recombinant proteins. The authors focus on two areas of interest: the use of E. coli vectors and strains for production of pure, functional protein, and the use of E. coli as host for the functional screening of large collections of proteins and peptides. Among the cutting-edge techniques demonstrated are those for rapid high-level expression and purification of soluble and functional recombinant protein and those essential to functional genomics, proteomics, and protein engineering.

Recenzje

"...many will find it extremely useful to have this book on hand when trying to achieve high levels of pure, soluble recombinant protein; or incorporating an E. coli expression step into the procedures of functional genomics, proteomics and protein engineering." - Microbiology Today

Spis treści

Cold-Inducible Promoters for Heterologous Protein Expression François Baneyx and Mirna Mujacic Dual-Expression Vectors for Efficient Protein Expression in Both E. coli and Mammalian Cells Rebecca L. Mullinax, David T. Wong, Heidi A. Davis, Kerstein A. Padgett, and Joseph A. Sorge A Dual-Expression Vector Allowing Expression in E. coli and P. pastoris, Including New Modifications Angelika Lueking, Sabine Horn, Hans Lehrach, and Dolores J. Cahill Purification of Recombinant Proteins from E. coli by Engineered Inteins Ming-Qun Xu and Thomas C. Evans, Jr Calmodulin as an Affinity Purification Tag Samu Melkko and Dario Neri Calmodulin-Binding Peptide as a Removable Affinity Tag for Protein Purification Wolfgang Klein Maltose-Binding Protein as a Solubility Enhancer Jeffrey D. Fox and David S. Waugh Thioredoxin and Related Proteins as Multifunctional Fusion Tags for Soluble Expression in E. coli Edward R. LaVallie, Elizabeth A. DiBlasio-Smith, Lisa A. Collins-Racie, Zhijian Lu, and John M. McCoy Discovery of New Fusion Protein Systems Designed to Enhance Solubility in E. coli Gregory D. Davis and Roger G. Harrison Assessment of Protein Folding/Solubility in Live Cells Rhesa D. Stidham, W. Christian Wigley, John F. Hunt, and Philip J. Thomas Improving Heterologous Protein Folding via Molecular Chaperone and Foldase Co-Expression François Baneyx and Joanne L. Palumbo High-Throughput Purification of PolyHis-Tagged Recombinant Fusion Proteins Thomas Lanio, Albert Jeltsch, and Alfred Pingoud Co-Expression of Proteins in E. coli Using Dual Expression Vectors Karen Johnston and Ronen Marmorstein Small-Molecule Affinity-Based Matrices for Rapid Protein Purification Karin A. Hughes and Jean P. Wiley Use of tRNA-Supplemented Host Strains for Expression ofHeterologous Genes in E. coli Carsten-Peter Carstens Screening Peptide/Protein Libraries Fused to the l Repressor DNA-Binding Domain in E. coli Cells Leonardo Mariño-Ramírez, Lisa Campbell, and James C. Hu Studying Protein-Protein Interactions Using a Bacterial Two-Hybrid System Simon L. Dove Using Bio-Panning of FLITRX Peptide Libraries Displayed on E. coli Cell Surface to Study Protein-Protein Interactions Zhijian Lu, Edward R. LaVallie, and John M. McCoy Use of Inteins for the In Vivo Production of Stable Cyclic Peptide Libraries in E. coli Ernesto Abel-Santos, Charles P. Scott, and Stephen J. Benkovic Hyperphage: Improving Antibody Presentation in Phage Display Olaf Broders, Frank Breitling, and Stefan Dübel Combinatorial Biosynthesis of Novel Carotenoids in E. coli Gerhard Sandmann Using Transcriptional-Based Systems for In Vivo Enzyme Screening Steven M. Firestine, Frank Salinas, and Stephen J. Benkovic Identification of Genes Encoding Secreted Proteins Using Mini-OphoA Mutagenesis Mary N. Burtnick, Paul J. Brett, and Donald E. Woods Index

Blurb

Gene expression using E. coli as a host is carried out over a wide range of disciplines in academic and industrial laboratories. In E. coli Gene Expression Protocols, Peter E. Vaillancourt presents a collection of popular and emerging methodologies that take advantage of E. coli's ability to quickly and inexpensively express recombinant proteins. The authors focus on two areas of interest: the use of
E. coli vectors and strains for production of pure, functional protein, and the use of E. coli as host for the functional screening of large collections of proteins and peptides. Among the cutting-edge techniques demonstrated are those for rapid high-level expression and purification of soluble and functional recombinant protein, and those essential to functional genomics, proteomics, and protein engineering. Described in step-by-step detail to ensure robust, trouble-free results, each proven method has been written by a hands-on expert and includes extensive notes and practical tips for avoiding pitfalls. Even highly skilled researchers will find many time-saving techniques.
Authoritative and highly practical, E. coli Gene Expression Protocols provides a state-of-the-art collection of tested methods for this powerful gene expression technology, offering today's investigators proven tools for success in the emerging fields of functional genomics and proteomics.