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This is a unique book that describes the most recent achievements in the methodology of protein biosynthesis under cell-free conditions. Various versions of cell-free protein-synthesizing systems and their applications to production of individual proteins on a preparative scale are reviewed. The most recent, advanced methodologies, such as continuous-exchange and continuous-flow cell-free systems and novel effecting batch-format cell-free procedures, are considered. Special attention is drawn to the possibilities of structural (NMR; X-ray) analysis of various gene expression products with the use of a new generation of cell-free systems.
List of contents
I Introduction.- 1 Cell-Free Protein Synthesis.- II Improved Classical (Batch) Cell-Free Translation Systems.- 2 Improved Batch Translation System Based on E. coli Extract.- 3 Improved Composition and Energy Supply for Bacterial Batch Systems.- 4 PURE System: New Cell-Free Translation System Reconstructed with Purified Components.- 5 Direct Expression of PCR Products in Cell-Free Translation Systems.- 6 Yeast Cell-Free Translation Systems.- 7 Poly(A)-Dependent Cell-Free Translation Systems from Animal Cells.- III Continuous-Flow and Continuous-Exchange Cell-Free Translation Systems.- 8 Continuous-Flow and Continuous-Exchange Cell-Free Translation Systems and Reactors.- 9 Highly Productive Plant Continuous Cell-Free System.- 10 Matrix Reactor: A New Scalable Reactor Principle for Cell-Free Protein Expression.- IV Protein Folding in Cell-Free Translation System.- 11 Co-Translational Protein Folding in Prokaryotic and Eukaryotic Cell-Free Translation Systems.- 12 Expression of an Aggregation-Prone Protein in the RTS 500 System.- 13 Expression of Soluble Murine Endostatin with the RTS System.- 14 Cell-Free Expression of Soluble Human Erythropoietin.- 15 Cell-Free Expression of a 127 kDa Protein: The Catalytic Subunit of Human Telomerase.- 16 The Expression of Disulfide Bonded Proteins in Cell-Free Protein Expression.- V Special Topics and Short Communications.- 17 Structural and Functional Compensation by Proteins for the RNA Deficit of Animal Mitochondrial Translation Systems.- 18 Proficient Target Selection in Structural Genomics by In Vitro Protein Expression on Gateway Recombination Plasmids.- 19 High-Level Cell-Free Protein Expression from PCR-Generated DNA Templates.- 20 Optimization of the Translation Initiation Region of Prokaryotic Expression Vectors: High Yield InVitro Protein Expression and mRNA Folding.- 21 Selective Labeling of Proteins in the RTS 500 System.- 22 In Vitro Protein Production for Structure Determination with the RTS System.- 23 Sequence Specific Biotinylation and Purification of Proteins Expressed in the RTS 500 System.- 24 Synthesis of the His 6-Tagged Recombinant Protein APPC99-His and His-PS1 Using RTS 500.
