E. Coli Gene Expression Protocols Hardback - 2002

Rapid transfer of exponentially growing E. coli cultures from physiological to low temperatures (10-15 C) has profound consequences on cell physiology: membrane fluidity decreases, which interferes with transport and secretion, the secondary structures of nucleic acids are stabilized, which affect the efficiencies of mRNA transcription/translation and DNA replication, and free ribosomal sub-units and 70S particles accumulate at the expense of polysomes, negatively impacting translation of most cellular mRNAs (1-3).

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.