1st Edition

Cell-Free Protein Expression

By W. Antoni Kudlicki Copyright 2007
    242 Pages 93 B/W Illustrations
    by CRC Press

    Following its inception in the 1950s, cell-free protein synthesis made a tremendous impact on the basic life sciences. The use of cell-free systems was key to understanding molecular mechanisms underlying one of the most complicated processes found in nature: protein translation. Since this time, aggressive cutting-edge research and stiff commerical competition have driven the development of a variety of systems with increased productivity, improved protein quality and relatively low production costs. As a result, technology has generated myriad applications that have enabled advances in fields as diverse as systems biology, structural biology, and drug discovery. Cell-Free Protein Expression describes and expands upon many of these applications. The volume has been divided into six main sections. In the first section, many of the most popular sources of cell-free lysates are introduced. The second section focuses on extraordinary advances made in the Escherichia coli-based systems that have enabled reconstitution of the entire translational process, incorporation of post-translational modifications, yield increase, and production of functional membrane proteins. This progress extends the usefulness of cell-free systems into structural biology applications described in the third section and high-content platforms like protein microarrays discussed in the fourth section. The final two sections cover the use of cell-free protein expression technologies in the rational design and directed evolution of proteins within the scientific community.

    Nemetz Generation of Linear Expression Elements by PCR Nemetz et al. Rapid Protein Engineering by Expression-PCR Stiege Expression-PCR: From Gene Pools to Purified Proteins within One Day Watzele High-throughput expression PCR Used to Systematically Investigate Regulation of Translation Initiation in an E. coli Cell-Free Expression System Grantzdorffer Reduction of Primer-Dimer Formation During Generation of Expression Fragments by PCR Pelton Isotope Labeling of Proteins for NMR Spectroscopy Using Cell-Free Methods Fernholz Production of a Specifically Labeled Protein in mg Quantities for NMR Analysis Schlossmann In Situ Mono-Biotinylation of Cell-Free Expressed Proteins Using the AviTag Technology Schraml Rapid Generation of Protein Variants and Subsequent Analysis by Surface Plasmon Resonance Bernhard Incorporation of Fluorescence Labels into Cell-Free Produced Proteins Budisa Expression of 'tailor-made' proteins via Incorporation of Synthetic Amino Acids by Using Cell-Free Protein Synthesis Beernink Application of Cell-Free Expression Systems to Proteomic Studies Busso/Kim Screening and Increasing Soluble Expression of Recombinant Proteins using the RTS 100 HY in a 96-Well format Yamane In vitro expression of proteins with disulfide bridges and ist application for high-throughput screening system Kim Cell-free expression of proteins containing multiple disulfide bonds Maurer Cell-free Synthesis of Membrane Proteins on a Preparative Scale Betton Using Maltose-Binding Protein Fragment Complementation to Probe Protein-Protein Interactions by Co-Expression in the RTS System Lorenz In-vitro translation of KRAB Zinc Finger Transcriptional Repressor Proteins and their Interaction withtheir TIF1ss Co-Repressor Lehmann Optimization of Cell-Free Expression of FAD-dependent D-Amino Acid Oxidase Liu Expression of Recombinant Chemokine-like Factor 1 with a Cell-Free Protein Biosynthesis System Bosserhoff Recombinant Expression of Functional Active MIA Protein for Mutation Analysis Using RTS System Bochtler-Hock Cell-Free Expression of the Heterodimeric Protein Penicillin G Amidase in a Functional Active Form Bittorf Cell-Free Expression of the His-Tagged Recombinant Prolactin-like Placenta Protein E Using the RTS 500 System Iskakov Complementary Interaction Between the Central Domain of 18S rRNA and the 5'Untranslated Region of mRNA Enhances Translation Efficiency in Plants Kubick In vitro translation in an insect-based cell-free system


    Kudlicki, W. Antoni