05030nam 22006254a 450 991082988420332120230721033032.03-527-69150-21-281-94699-097866119469993-527-62270-53-527-62269-1(CKB)1000000000550884(EBL)481824(OCoLC)277152635(SSID)ssj0000119208(PQKBManifestationID)11144005(PQKBTitleCode)TC0000119208(PQKBWorkID)10057867(PQKB)10715635(MiAaPQ)EBC481824(EXLCZ)99100000000055088420090404d2008 uy 0engur|n|---|||||txtccrCell-free protein synthesis[electronic resource] methods and protocols /edited by Alexander S. Spirin and James R. SwartzWeinheim Wiley-VCHc20081 online resource (264 p.)Description based upon print version of record.3-527-31649-3 Includes bibliographical references and index.Cell-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)1.2.1.2 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 Regeneration1.6 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 System2.3.4 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 Templates3.2.3 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 β-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 Process4.2.1 Fusion-protein Production in the Cell-free SystemWith 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.ProteinsSynthesisGenetic translationProteinsSynthesis.Genetic translation.572.6Spirin A. S(Aleksandr Sergeevich)64527Swartz James R1713547MiAaPQMiAaPQMiAaPQBOOK9910829884203321Cell-free protein synthesis4106601UNINA01620oam 2200301z- 450 991016331710332120230913112557.01-62380-332-2(CKB)3710000001046549(BIP)055283754(Exl-AI)993710000001046549(EXLCZ)99371000000104654920210505c2013uuuu -u- -engCluelessHarmony Ink Press1 online resource (124 p.) Keeping Secrets: Book TwoOut jock Tommy Johnson and former bully Jason Strummer have settled into a surprisingly comfortable routine. They're even planning a special camping trip for just the two of them for their one-month anniversary. Tommy doesn't think things can get any better--until Jason's past catches up to them.Jason's stepdad will stop at nothing to put Jason back in his place. As Jason spins out of control, Tommy tries desperately to hold the edges of their world together. But the pressure of dealing with Jason's increasingly erratic behavior and keeping a growing number of secrets under wraps may prove too much for Tommy to handle.Then Jason blows it and their comfy world falls apart. When he finally texts Tommy for help, they both learn some secrets aren't worth keeping.Gay teenagersFictionGenerated by AIInterpersonal relationsFictionGenerated by AIGay teenagersInterpersonal relationsRoman J371177Cain AnneothBOOK9910163317103321Clueless3593325UNINA