Cell-free protein synthesis [[electronic resource] ] : methods and protocols / / edited by Alexander S. Spirin and James R. Swartz

• Record Nr.: UNINA9910144101803321
• Pubbl/distr/stampa: Weinheim, : Wiley-VCH, c2008
• ISBN: 3-527-69150-2; 1-281-94699-0; 9786611946999; 3-527-62270-5; 3-527-62269-1
• Descrizione fisica: 1 online resource (264 p.)
• Altri autori (Persone): SpirinA. S (Aleksandr Sergeevich); SwartzJames R
• Disciplina: 572.6
• Soggetti: Proteins - Synthesis; Genetic translation; Electronic books.
• Lingua di pubblicazione: Inglese
• Formato: Materiale a stampa
• Livello bibliografico: Monografia
• Note generali: Description based upon print version of record.
• Nota di bibliografia: Includes bibliographical references and index.
• Nota di contenuto: 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 Regeneration; 1.6 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; 2.3.4 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; 3.2.3 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 Process; 4.2.1 Fusion-protein Production in the Cell-free System
• Sommario/riassunto: With 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.

Feel the Grass Grow : Ecologies of Slow Peace in Colombia

• Record Nr.: UNINA9910860869703321
• Autore: Lederach Angie
• Pubbl/distr/stampa: Redwood City, : Stanford University Press, , 2023; ©2023
• ISBN: 9781503635692; 9781503635685
• Edizione: [1st ed.]
• Descrizione fisica: 1 online resource (287 pages)
• Disciplina: 303.6609861
• Soggetti: Peace-building - Colombia; Peasants - Political activity - Colombia; Political violence - Colombia
• Lingua di pubblicazione: Inglese
• Formato: Materiale a stampa
• Livello bibliografico: Monografia
• Nota di contenuto: Cover -- Title Page -- Copyright -- Dedication -- Contents -- Acknowledgments -- List of Acronyms -- To Defend Life: An Introduction -- Part I. Memorias Vivas-Living Memories -- 1. From and For the Territory: The Campesino Struggle for Peace -- 2. The Earth Suffered, Too: The Death of the Avocado Forest and Multispecies Resurgence -- Part II. Prisa-Hurry -- 3. Photos and Signatures: Contested Performances of Peace -- 4. Too Much Prisa: The Temporal Dynamics of Violence and Peace -- Part III. Paz Sin Prisa-Slow Peace -- 5. The Times of Slow Peace -- 6 Voice and Votes: Building Territorial Peace -- 7. Vigías of Hope: Slow Peace and the Ethics of Attention -- Coda -- Notes -- References -- Index -- Back Cover.
• Sommario/riassunto: "On November 24, 2016, the Colombian government and the Revolutionary Armed Forces of Colombia signed a revised peace accord that marked a political end to over a half-century of war. Feel the Grass Grow traces the far less visible aspects of moving from war to peace: the decades of campesino struggle to defend life, land, and territory prior to the national accord, as well as campesino social leaders' engagement with the challenges of the state's post-accord reconstruction efforts. In the words of the campesino peace activists, "peace is not signed, peace is built." Drawing on nearly a decade of ; extensive ethnographic and participatory research, Angela Jill Lederach advances a theory of "slow peace." Slowing down does not negate the urgency that animates the defense of territory in the context of the interlocking processes of political and environmental violence that persist in post-accord Colombia. Instead, Lederach shows how the campesino call to "slowness" recenters grassroots practices of peace, grounded in multigenerational struggles for territorial liberation. In examining the various layers of meaning embedded within campesino theories of "the times (los tiempos)," this book directs analytic attention to the holistic understanding of peacebuilding found among campesino social leaders. Their experiences of peacebuilding shape an understanding of time as embodied, affective, and emplaced. The call to slow peace gives primacy to the everyday, where relationships are deepened, ancestral memories reclaimed, and ecologies regenerated"--