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Manipulation of recombinant DNA, which is almost exclusively performed using the host E. coli, constitutes one of the fundamental methodologies of molecular biotechnology. In E. coli Plasmid Vectors, experienced bench researchers describe their proven techniques for the manipulation of recombinant plasmids utilizing this popular bacterial host. The authors describe readily reproducible methods for cloning DNA into plasmid vectors, transforming plasmids into E. coli, and analyzing recombinant clones. They also include protocols for the construction and screening of libraries, as well as specific techniques for specialized cloning vehicles, such as cosmids, bacterial artificial chromosomes, l vectors, and phagemids. Also presented are methods for common downstream applications, such as mutagenesis, expression of recombinant proteins and RNA transcripts, and uses of reporter genes. Each fully tested protocol is described in step-by-step detail by an established expert in the field and includes an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, tips on troubleshooting and avoiding known pitfalls and, where needed, a discussion of the interpretation and use of the results. Comprehensive and highly practical, E. coli Plasmid Vectors offers those new to the field a basic guide to the use of plasmid vectors in the cloning host E. coli, and those more experienced researchers a broad-ranging, proven array of successful techniques.
Plasmids --- Escherichia coli --- Genetic vectors --- Cloning vectors --- Expression vectors (Genetics) --- rDNA vectors --- Recombinant DNA vectors --- Vectors, Genetic --- Gene expression --- Molecular cloning --- Recombinant DNA --- E. coli (Bacterium) --- Escherichia --- Paragenes --- Cytoplasmic inheritance --- Mobile genetic elements --- Extrachromosomal DNA --- Biochemistry. --- Cytology. --- Biochemistry, general. --- Cell Biology. --- Cell biology --- Cellular biology --- Biology --- Cells --- Cytologists --- Biological chemistry --- Chemical composition of organisms --- Organisms --- Physiological chemistry --- Chemistry --- Medical sciences --- Composition
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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.
Escherichia coli --- Gene expression --- Genetics --- E. coli (Bacterium) --- Escherichia --- Genes --- Genetic regulation --- Expression --- Biochemistry. --- Cytology. --- Biochemistry, general. --- Cell Biology. --- Cell biology --- Cellular biology --- Biology --- Cells --- Cytologists --- Biological chemistry --- Chemical composition of organisms --- Organisms --- Physiological chemistry --- Chemistry --- Medical sciences --- Composition --- Escherichia coli - Genetics
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