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Protein Engineering

Editat de Uwe T. Bornscheuer, Matthias Höhne
en Limba Engleză Hardback – 30 oct 2017
This volume details basic and advanced protocols for both stages of protein engineering: the library design phase and the identification of improved variants by screening and selection. Chapters focus on enzyme engineering using rational and semi-rational approaches. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Protein Engineering: Methods and Protocols aims to aid scientists in the planning and performance of their experiments. 
The chapter 'Functional Analysis of Membrane Proteins Produced by Cell-Free Translation' is open access under a CC BY 4.0 license via link.springer.com. 


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Specificații

ISBN-13: 9781493973644
ISBN-10: 1493973649
Pagini: 364
Ilustrații: XI, 350 p. 82 illus., 63 illus. in color.
Dimensiuni: 183 x 260 x 26 mm
Greutate: 0.88 kg
Ediția:1st edition 2018
Editura: Humana
Locul publicării:New York, NY, United States

Cuprins

Protein Engineering - Past, Present, and Future.- (Semi)rational Protein Design.- Computational Analysis of Protein Tunnels and Channels.- YASARA - A Tool to Obtain Structural Guidance in Biocatalytic Investigations.- A Computational Library Design Protocol for Rapid Improvement of Protein Stability – FRESCO.- Directed Evolution of Proteins Based on Mutational Scanning.- A Brief Guide to the High-throughput Expression of Directed Evolution Libraries.- Library Growth and Protein Expression: Optimal and Reproducible Microtiter Plate Expression of Recombinant Enzymes in E. coli using MTP Shakers.- Normalized Screening of Protein Engineering Libraries by Split-GFP Crude Cell Extract Quantification.- Functional Analysis of Membrane Proteins Produced by Cell-Free Translation.- Practical Considerations Regarding the Choice of the Best High-throughput Assay.- High-throughput Screening Assays for Lipolytic Enzymes.- Continuous High-throughput Colorimetric Assays for α-transaminases.-Colorimetric High-troughput Screening Assays for the Directed Evolution of Fungal Laccases.- Directed Co-evolution of Two Cellulosic Enzymes in Escherichia coli Based on their Synergistic Reactions.- Program-guided Design of High-throughput Enzyme Screening Experiments and Automated Data Analysis/Evaluation.- Solid Phase Agar Plate Assay for Screening Amine Transaminases.- Ultrahigh-throughput Screening of Single-cell Lysates for Directed Evolution and Functional Metagenomics.- Isolation of pH-sensitive Antibody Fragments by Fluorescence-activated Cell Sorting and Yeast Surface Display.- Library Generation and Auxotrophic Selection Assays in Escherichia coli and Thermus thermophiles. 

Textul de pe ultima copertă

This volume details basic and advanced protocols for both stages of protein engineering: the library design phase and the identification of improved variants by screening and selection. Chapters focus on enzyme engineering using rational and semi-rational approaches. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Protein Engineering: Methods and Protocols aims to aid scientists in the planning and performance of their experiments. 

Caracteristici

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts