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.
Table of contents
Section One: TECHNOLOGIES
Chapter One - Energy Systems for ATP Regeneration in Cell-free Protein Synthesis Reactions
Kara A. Calhoun, James R. Swartz
Chapter 2 - Continuous-exchange Protein-synthesizing Systems
Vladimir A. Shirokov, Aigar Kommer, Vyacheslav A. Kolb, Alexander S. Spirin
Chapter 3 - Cell-free production of integral membrane proteins on a preparative scale"
Christian Klammt, Daniel Schwarz, Vilker Dötsch, Frank Bernhard
Chapter 4- SIMPLEX: Single-molecule PCR-linked in vitro expression: a novel method for high-throughput construction and screening of protein libraries
Suang Rungpragayphan, Tsuneo Yamane, Hideo Nakano
Chapter 5 - Methods for High-throughput Materialization of Genetic Information Based on Wheat Germ Cell-free Expression System
Tatsuya Sawasaki, Ryo Morishita, Mudeppa D. Gouda, Yaeta Endo
Chapter 6 - Exogenous Protein Expression in Xenopus Oocytes: Basic Procedures
Elena Bossi, Maria Serena Fabbrini, Aldo Ceriotti
Section Two: APPLICATIONS
Chapter 7 -In vitro translation to study HIV protease activity
Zene Matsuda, Mutsunori Iga, Kosuke Miyauchi, Jun Komano, Kazuhiro Morishita, Akihiko Okayama, Hirohito Tsubouchi
Chapter 8 - The protein truncation test in mutation detection and molecular diagnosis
Oliver Hauss, Oliver Müller
Chapter 9 - Creation of Novel Enantioselective lipases by SIMPLEX
Yuichi Koga, Tsuneo Yamane, Hideo Nakano
Chapter 10 - In vitro transcription and translation coupled to 2D-electrophoresis for bacterial proteome analysis
Nathalie Norais, Ignazio Garaguso, Germano Ferrari, Guido Grandi
Chapter 11 - In vitro Screen of Bioinformatically Selected Bacillus anthracis Vaccine Candidates by Coupled Transcription, Translation and Immunoprecipitation Analysis
Orit Gat, Haim Grosfeld, Avigdor Shafferman
Chapter 12 - Functional expression of type 1 rat GABA transporter in microinjected Xenopus laevis oocytes
Stefano Giovannardi, Simona Magagnin, Andrea Soragna, Laura Faravelli
Chapter 13 - Production of Protein for NMR Study Using the Wheat Germ Cell-Free System
Toshiyuki Kohno, Yaeta Endo