LEADER 05003nam 22006254a 450 001 9910876999303321 005 20200520144314.0 010 $a3-527-69150-2 010 $a1-281-94699-0 010 $a9786611946999 010 $a3-527-62270-5 010 $a3-527-62269-1 035 $a(CKB)1000000000550884 035 $a(EBL)481824 035 $a(OCoLC)277152635 035 $a(SSID)ssj0000119208 035 $a(PQKBManifestationID)11144005 035 $a(PQKBTitleCode)TC0000119208 035 $a(PQKBWorkID)10057867 035 $a(PQKB)10715635 035 $a(MiAaPQ)EBC481824 035 $a(EXLCZ)991000000000550884 100 $a20090404d2008 uy 0 101 0 $aeng 135 $aur|n|---||||| 181 $ctxt 182 $cc 183 $acr 200 00$aCell-free protein synthesis $emethods and protocols /$fedited by Alexander S. Spirin and James R. Swartz 210 $aWeinheim $cWiley-VCH$dc2008 215 $a1 online resource (264 p.) 300 $aDescription based upon print version of record. 311 $a3-527-31649-3 320 $aIncludes bibliographical references and index. 327 $aCell-free Protein Synthesis; Contents; Preface; List of Contributors; 1 Cell-free Protein Synthesis Systems: Historical Landmarks, Classification, and General Methods; 1.1 Introduction: Historical Landmarks; 1.1.1 Discovery of Protein Synthesis in Cell Extracts; 1.1.2 Translation of Exogenous Messages; 1.1.3 Coupled Transcription-translation in Bacterial Extracts; 1.1.4 Combined Transcription-translation Systems; 1.1.5 Continuous Flow/Continuous Exchange Principle; 1.2 Prokaryotic and Eukaryotic Types of Cell-free Expression Systems; 1.2.1 Cell Extracts; 1.2.1.1 E. coli extract (ECE) 327 $a1.2.1.2 Wheat Germ Extract (WGE)1.2.1.3 Rabbit Reticulocyte Lysate (RRL); 1.2.2 Genetic Constructs (Expression Vectors); 1.2.2.1 Prokaryotic Systems; 1.2.2.2 Eukaryotic Systems; 1.3 Preparing Cell Extracts; 1.3.1 E. coli Extracts; 1.3.1.1 Genetics; 1.3.1.2 Cell Growth; 1.3.1.3 Extract Preparation; 1.3.2 Wheat Germ Extracts; 1.4 Designing Reaction Composition; 1.4.1 Mg(2+) and Phosphate; 1.4.2 Other Salts; 1.4.3 Nucleotides and Amino Acids; 1.4.4 Stabilization Reagents; 1.4.5 Other Factors; 1.5 Providing Energy; 1.5.1 Direct Nucleotide Regeneration; 1.5.2 Indirect Nucleotide Regeneration 327 $a1.6 Enhancing Protein Folding1.6.1 Temperature Effects; 1.6.2 Cell Extract Concentration; 1.6.3 Effects of Folding Ligands; 1.6.4 Effects of Chaperones and Foldases; 1.6.5 Effects of Detergents; 2 The Constructive Approach for Cell-free Translation; 2.1 Introduction; 2.2 The Process of Protein Synthesis; 2.2.1 Polypeptide Synthesis; 2.2.2 Protein Maturation; 2.3 A Constructive Approach to Protein Synthesis; 2.3.1 In Vitro Reconstitution of Polypeptide Synthesis; 2.3.2 Protocol of Protein Synthesis using PURE System; 2.3.3 Addition of Protein Folding Machinery to the PURE System 327 $a2.3.4 Integration of a Membrane Targeting System with the PURE system2.3.5 Protein Synthesis using the PURE System containing Molecular Chaperones; 2.4 Conclusion; 3 Functional Genomic Analysis using Sequential Cell-free Protein Synthesis; 3.1 Introduction; 3.1.1 The Post-genomic Era; 3.1.2 Cell-free Protein Synthesis (CFPS) as a Functional Proteomic Tool; 3.2 Developing an enabling Technology for Sequential Expression Analysis; 3.2.1 Improving Linear Template Stability; 3.2.2 Improving PCR Reactions for generating Genomic Linear Templates 327 $a3.2.3 Optimizing Cofactor Concentrations for Enzyme Activation3.3 Demonstrating Functional Genomic Analysis with CFPS; 3.3.1 Isolation and Expression of Genomic Targets; 3.3.2 Effects of Sample Library on ?-Lactamase Expression and Activity; 3.4 Conclusions and Projections; 4 Cell-free Technology for Rapid Production of Patient-specific Fusion Protein Vaccines; 4.1 Introduction; 4.1.1 Lymphoma and Fusion Protein Vaccine Treatments; 4.1.2 Comparing Cell-free and In Vivo Production Systems; 4.2 Developing the Fusion Protein Construct and the Cell-free Production Process 327 $a4.2.1 Fusion-protein Production in the Cell-free System 330 $aWith its detailed description of membrane protein expression, high-throughput and genomic-scale expression studies, both on the analytical and the preparative scale, this book covers the latest advances in the field. The step-by-step protocols and practical examples given for each method constitute practical advice for beginners and experts alike. 606 $aProteins$xSynthesis 606 $aGenetic translation 615 0$aProteins$xSynthesis. 615 0$aGenetic translation. 676 $a572.6 701 $aSpirin$b A. S$g(Aleksandr Sergeevich)$064527 701 $aSwartz$b James R$01761589 801 0$bMiAaPQ 801 1$bMiAaPQ 801 2$bMiAaPQ 906 $aBOOK 912 $a9910876999303321 996 $aCell-free protein synthesis$94201136 997 $aUNINA