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.

Section One: TECHNOLOGIESChapter One - Energy Systems for ATP Regeneration in Cell-free Protein Synthesis ReactionsKara A. Calhoun, James R. SwartzChapter 2 - Continuous-exchange Protein-synthesizing SystemsVladimir A. Shirokov, Aigar Kommer, Vyacheslav A. Kolb, Alexander S. SpirinChapter 3 - Cell-free production of integral membrane proteins on a preparative scale'Christian Klammt, Daniel Schwarz, Vilker Dötsch, Frank BernhardChapter 4- SIMPLEX: Single-molecule PCR-linked in vitro expression: a novel method for high-throughput construction and screening of protein librariesSuang Rungpragayphan, Tsuneo Yamane, Hideo NakanoChapter 5 - Methods for High-throughput Materialization of Genetic Information Based on Wheat Germ Cell-free Expression SystemTatsuya Sawasaki, Ryo Morishita, Mudeppa D. Gouda, Yaeta EndoChapter 6 - Exogenous Protein Expression in Xenopus Oocytes: Basic ProceduresElena Bossi, Maria Serena Fabbrini, Aldo CeriottiSection Two: APPLICATIONSChapter 7 -In vitro translation to study HIV protease activityZene Matsuda, Mutsunori Iga, Kosuke Miyauchi, Jun Komano, Kazuhiro Morishita, Akihiko Okayama, Hirohito TsubouchiChapter 8 - The protein truncation test in mutation detection and molecular diagnosisOliver Hauss, Oliver MüllerChapter 9 - Creation of Novel Enantioselective lipases by SIMPLEXYuichi Koga, Tsuneo Yamane, Hideo NakanoChapter 10 - In vitro transcription and translation coupled to 2D-electrophoresis for bacterial proteome analysisNathalie Norais, Ignazio Garaguso, Germano Ferrari, Guido GrandiChapter 11 - In vitro Screen of Bioinformatically Selected Bacillus anthracis Vaccine Candidates by Coupled Transcription, Translation and Immunoprecipitation AnalysisOrit Gat, Haim Grosfeld,Avigdor ShaffermanChapter 12 - Functional expression of type 1 rat GABA transporter in microinjected Xenopus laevis oocytesStefano Giovannardi, Simona Magagnin, Andrea Soragna, Laura FaravelliChapter 13 - Production of Protein for NMR Study Using the Wheat Germ Cell-Free SystemToshiyuki Kohno, Yaeta Endo