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.

Cold-Inducible Promoters for Heterologous Protein Expression.- Dual-Expression Vectors for Efficient Protein Expression in Both E. coli and Mammalian Cells.- A Dual-Expression Vector Allowing Expression in E. coli and P. pastoris, Including New Modifications.- Purification of Recombinant Proteins from E. coli by Engineered Inteins.- Calmodulin as an Affinity Purification Tag.- Calmodulin-Binding Peptide as a Removable Affinity Tag for Protein Purification.- Maltose-Binding Protein as a Solubility Enhancer.- Thioredoxin and Related Proteins as Multifunctional Fusion Tags for Soluble Expression in E. coli.- Discovery of New Fusion Protein Systems Designed to Enhance Solubility in E. coli.- Assessment of Protein Folding/Solubility in Live Cells.- Improving Heterologous Protein Folding via Molecular Chaperone and Foldase Co-Expression.- High-Throughput Purification of PolyHis-Tagged Recombinant Fusion Proteins.- Co-Expression of Proteins in E. coli Using Dual Expression Vectors.- Small-Molecule Affinity-Based Matrices for Rapid Protein Purification.- Use of tRNA-Supplemented Host Strains for Expression of Heterologous Genes in E. coli.- Screening Peptide/Protein Libraries Fused to the ? Repressor DNA-Binding Domain in E. coli Cells.- Studying Protein-Protein Interactions Using a Bacterial Two-Hybrid System.- Using Bio-Panning of FLITRX Peptide Libraries Displayed on E. coli Cell Surface to Study Protein-Protein Interactions.- Use of Inteins for the In Vivo Production of Stable Cyclic Peptide Libraries in E. coli.- Hyperphage.- Combinatorial Biosynthesis of Novel Carotenoids in E. coli.- Using Transcriptional-Based Systems for In Vivo Enzyme Screening.- Identification of Genes Encoding Secreted Proteins Using Mini-OphoA Mutagenesis.