A highly anticipated update of the previous edition, In Vitro Transcription and Translation Protocols, Second Edition, provides molecular biology laboratories with the most powerful techniques for exploiting in vitro transcription and translation systems.In Section One: Technologies, authors discuss the use of alternative energy systems for ATP regeneration, the efficient in vitro production of integral membrane proteins, and the high-throughput production of protein libraries using the innovative single-molecule PRC-linked in vitro expression (SIMPLEX) technology. In Section Two: Applications, authors present in vitro translation of amplified protease gene from HIV patients to select proper anti-protease drug therapy, in vitro translation of genes from human patients to diagnose biologically relevant gene mutations, use of SIMPLEX technology to select enantioselective lipases from libraries of randomly produced lipase mutants, and the in vitro transcription and translation to identify proteins in 2D-maps.

This book is a highly anticipated update of the previous edition. In Vitro Transcription and Translation Protocols, Second Edition, provides molecular biology laboratories with the most powerful techniques for exploiting in vitro transcription and translation systems. Completely updated with new chapters and topics, there is in-depth analysis of current technologies and applications of in-vitro transcriptions and translations systems. Detailed protocols for protein production using different in vitro transcription and translation systems are included.

Technologies.- Energy Systems for ATP Regeneration in Cell-Free Protein Synthesis Reactions.- Continuous-Exchange Protein-Synthesizing Systems.- Cell-Free Production of Integral Membrane Proteins on a Preparative Scale.- SIMPLEX: Single-Molecule PCR-Linked In Vitro Expression.- Methods for High-Throughput Materialization of Genetic Information Based on Wheat Germ Cell-Free Expression System.- Exogenous Protein Expression in Xenopus Oocytes.- Applications.- In Vitro Translation to Study HIV Protease Activity.- The Protein Truncation Test in Mutation Detection and Molecular Diagnosis.- Creation of Novel Enantioselective Lipases by SIMPLEX.- In Vitro Transcription and Translation Coupled to Two-Dimensional Electrophoresis for Bacterial Proteome Analysis.- In Vitro Screen of Bioinformatically Selected Bacillus anthracis Vaccine Candidates by Coupled Transcription, Translation, and Immunoprecipitation Analysis.- Functional Expression of Type 1 Rat GABA Transporter in Microinjected Xenopus laevis Oocytes.- Production of Protein for Nuclear Magnetic Resonance Study Using the Wheat Germ Cell-Free System.