Directed Evolution

Methods and Protocols

Andrew Currin, Neil Swainston (Herausgeber)

Buch | Hardcover

278 Seiten

2022 | 1st ed. 2022
Springer-Verlag New York Inc.
978-1-0716-2151-6 (ISBN)

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This volume explores the latest techniques used by researchers to study directed evolution (DE) at each stage of the Design-Build-Test-Learn cycle. Chapters in this book cover topics such as designing overlap extension PCR primers for protein mutagenesis; antha-guided automation of Darwin assembly for the construction of bespoke gene libraries; rapid cloning of random mutagenesis libraries using PTO-Quickstep; and DE of glycosyltransferases by a single-cell screening method. 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.

Cutting-edge and comprehensive, Directed Evolution: Methods and Protocols is a valuable resource for scientists and researchers who are interested in learning more about this field and incorporating these studies into new experimental workflows.

Designing Overlap Extension PCR Primers for Protein Mutagenesis: A Programmatic Approach.- Recombination of Single Beneficial Substitutions Obtained from Protein Engineering by Computer-Assisted Recombination (CompassR).- Non-Degenerate Saturation Mutagenesis: Library Construction and Analysis via MAX and ProxiMAX Randomization.- Antha-Guided Automation of Darwin Assembly for the Construction of Bespoke Gene Libraries.- SpeedyGenesXL An Automated, High-Throughput Platform for the Preparation of Bespoke Ultra-Large Variant Libraries for Directed Evolution.- Facile Assembly of Combinatorial Mutagenesis Libraries using Nicking Mutagenesis.- GeneORator: An Efficient Method for the Systematic Mutagenesis of Entire Genes.- Rapid Cloning of Random Mutagenesis Libraries using PTO-QuickStep.- Construction of Strong Promoters by Assembling Sigma Factor Binding Motifs.- Application of Restriction Free (RF) Cloning in Circular Permutation.- Site-Directed Mutagenesis Method Mediated by Cas9.- Directed Evolution of Transcription Factor-Based Biosensors for Altered Effector Specificity.- A Screening Method for P450 BM3 Mutant Libraries using Multiplexed Capillary Electrophoresis for Detection of Enzymatically Converted Compounds.- Directed Evolution of Glycosyltransferases by a Single-Cell Ultrahigh-Throughput FACS-Based Screening Method.- Learning Strategies in Protein Directed Evolution.

Erscheinungsdatum	24.06.2022
Reihe/Serie	Methods in Molecular Biology ; 2461
Zusatzinfo	58 Illustrations, color; 5 Illustrations, black and white; XI, 278 p. 63 illus., 58 illus. in color. With online files/update.
Verlagsort	New York, NY
Sprache	englisch
Maße	178 x 254 mm
Themenwelt	Studium ► 2. Studienabschnitt (Klinik) ► Humangenetik
	Naturwissenschaften ► Biologie ► Genetik / Molekularbiologie
	Technik ► Umwelttechnik / Biotechnologie
Schlagworte	computationally guided protein stapling • de novo protein design • gene synthesis • Mutagenesis • SelFi
ISBN-10	1-0716-2151-3 / 1071621513
ISBN-13	978-1-0716-2151-6 / 9781071621516
Zustand	Neuware