E. coli Gene Expression Protocols

1. Auflage. Softcover version of original hardcover Auflage 2003
ISBN: 978-1-61737-302-2
Verlag: Humana Press

Buch, Englisch, Band 205, 347 Seiten, Previously published in hardcover, Format (B × H): 152 mm x 229 mm, Gewicht: 527 g

Reihe: Methods in Molecular Biology

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.

Zielgruppe

Research

Autoren/Hrsg.

Vaillancourt, Peter E.

Fachgebiete

• Naturwissenschaften Biowissenschaften Zellbiologie
• Medizin | Veterinärmedizin Medizin | Public Health | Pharmazie | Zahnmedizin Medizinische Fachgebiete Medizinische Mikrobiologie & Virologie
• Naturwissenschaften Biowissenschaften Bakteriologie
• Naturwissenschaften Biowissenschaften Mikrobiologie
• Naturwissenschaften Biowissenschaften Biowissenschaften Genetik und Genomik (nichtmedizinisch)
• Naturwissenschaften Biowissenschaften Molekularbiologie

Inhaltsverzeichnis

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 of Heterologous 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