Cell-Free Translation Systems

ISBN-13: 9783642639562 / Angielski / Miękka / 2012 / 259 str.

A. S. Spirin

Opis

CHAPTER 1 Cell-Free Protein Synthesis 3 ALEXANDER S. SPIRIN CHAPTER 1 Cell-Free Protein Synthesis 1 ALEXANDER S. SPIRIN* Abbreviations NTP nucleoside triphosphates aminoacyl-tRNA synthetases ARSes PEP phosphoenol pyruvate CP creatine phosphate AcP acetyl phosphate PK pyruvate kinase CK creatine kinase AcK acetyl kinase ME mercaptoethanol DTT dithiotreitol DHFR dihydrofolate reductase CAT chloramphenicol acetyltransferase GFP green fluorescent protein IL-2 and IL-6 interleukin-2 and interleukin-6 TMV tobacco mosaic virus Introduction: Prehistory of Cell-Free Translation Systems As early as the beginning of the 1950s, several groups independently demonstrat ed that protein synthesis does not require the integrity of the cell and can contin ue after cell disruption. Thus, disrupted cells or their isolated fractions were reported to be capable of synthesizing proteins (Borsook 1950; Winnick 1950a, 1950b; Siekevitz and Zamecnik 1951; Siekevitz 1952; Peterson and Greenberg 1952; Khesin 1953; Gale and Folkes 1954). In the meantime, ribonucleoprotein particles were observed and identified in cells (Palade 1955) and then isolated from cells and studied with respect to their physicochemical properties (Chao and Schachman 1956; Ts'o et al. 1956; Peterman and Hamilton 1957; Tissieres and Watson 1958; Tissieres et al. 1959; see also papers in Roberts 1958). The protein- * Institute of Protein Research, Russian Academy of Sciences, 142290 Pushchino, Moscow Region, Russia Tel. Fax: 007(095)924-0493, e-mail: spirin@vega. protres. ru Introduction synthesizing ability of these particles was experimentally proved (Littlefield et al. 1955; Littlefield and Keller 1957; McQuillen et al. 1959). The word "ribosome" was proposed to designate the protein-synthesizing ribonucleoprotein particles."

Spis treści

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 In Vitro 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.

Blurb

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.