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Directed Evolution

Methods and Protocols

  • Book
  • © 2022

Overview

Editors:
  1. Andrew Currin

    1. Manchester Institute of Biotechnology, University of Manchester, Manchester, UK

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    You can also search for this editor in PubMed   Google Scholar

  2. Neil Swainston

    1. Manchester Institute of Biotechnology, University of Manchester, Manchester, UK

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    You can also search for this editor in PubMed   Google Scholar

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

Part of the book series: Methods in Molecular Biology (MIMB, volume 2461)

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Table of contents (16 protocols)

  1. Front Matter

    Pages i-xi
    Download chapter PDF

  2. Designing Overlap Extension PCR Primers for Protein Mutagenesis: A Programmatic Approach

    • Xiaofang Huang, Liangting Xu, Chuyun Bi, Lili Zhao, Limei Zhang, Xuanyang Chen et al.
    Pages 1-7

  3. Recombination of Single Beneficial Substitutions Obtained from Protein Engineering by Computer-Assisted Recombination (CompassR)

    • Haiyang Cui, Mehdi D. Davari, Ulrich Schwaneberg
    Pages 9-18

  4. Nondegenerate Saturation Mutagenesis: Library Construction and Analysis via MAX and ProxiMAX Randomization

    • Anupama Chembath, Ben P. G. Wagstaffe, Mohammed Ashraf, Marta M. Ferreira Amaral, Laura Frigotto, Anna V. Hine
    Pages 19-41

  5. Antha-Guided Automation of Darwin Assembly for the Construction of Bespoke Gene Libraries

    • P. Handal-Marquez, M. Koch, D. Kestemont, S. Arangundy-Franklin, V. B. Pinheiro
    Pages 43-66

  6. SpeedyGenesXL: an Automated, High-Throughput Platform for the Preparation of Bespoke Ultralarge Variant Libraries for Directed Evolution

    • Joanna C. Sadler, Neil Swainston, Mark S. Dunstan, Andrew Currin, Douglas B. Kell
    Pages 67-83

  7. Facile Assembly of Combinatorial Mutagenesis Libraries Using Nicking Mutagenesis

    • Monica B. Kirby, Timothy A. Whitehead
    Pages 85-109

  8. GeneORator: An Efficient Method for the Systematic Mutagenesis of Entire Genes

    • Lucy Green, Nigel S. Scrutton, Andrew Currin
    Pages 111-122

  9. Rapid Cloning of Random Mutagenesis Libraries Using PTO-QuickStep

    • Pawel Jajesniak, Kang Lan Tee, Tuck Seng Wong
    Pages 123-135

  10. Construction of Strong Promoters by Assembling Sigma Factor Binding Motifs

    • Yonglin Zhang, Yang Wang, Jianghua Li, Chao Wang, Guocheng Du, Zhen Kang
    Pages 137-147

  11. Application of Restriction Free (RF) Cloning in Circular Permutation

    • Boudhayan Bandyopadhyay, Yoav Peleg
    Pages 149-163

  12. Site-Directed Mutagenesis Method Mediated by Cas9

    • Wanping Chen, Wenwen She, Aitao Li, Chao Zhai, Lixin Ma
    Pages 165-174

  13. Directed Evolution of Transcription Factor-Based Biosensors for Altered Effector Specificity

    • Leopoldo Ferreira Marques Machado, Neil Dixon
    Pages 175-193

  14. A Screening Method for P450 BM3 Mutant Libraries Using Multiplexed Capillary Electrophoresis for Detection of Enzymatically Converted Compounds

    • Anna Gärtner, Gustavo de Almeida Santos, Anna Joëlle Ruff, Ulrich Schwaneberg
    Pages 195-210

  15. Directed Evolution of Glycosyltransferases by a Single-Cell Ultrahigh-Throughput FACS-Based Screening Method

    • Yumeng Tan, Xue Zhang, Yan Feng, Guang-Yu Yang
    Pages 211-224

  16. Learning Strategies in Protein Directed Evolution

    • Xavier F. Cadet, Jean Christophe Gelly, Aster van Noord, Frédéric Cadet, Carlos G. Acevedo-Rocha
    Pages 225-275

  17. Correction to: Learning Strategies in Protein Directed Evolution

    • Xavier F. Cadet, Jean Christophe Gelly, Aster van Noord, Frédéric Cadet, Carlos G. Acevedo-Rocha
    Pages C1-C1

  18. Back Matter

    Pages 277-278
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Keywords

About this book

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.


Editors and Affiliations

  • Manchester Institute of Biotechnology, University of Manchester, Manchester, UK

    Andrew Currin, Neil Swainston

Bibliographic Information

  • Book Title: Directed Evolution

  • Book Subtitle: Methods and Protocols

  • Editors: Andrew Currin, Neil Swainston

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-0716-2152-3

  • Publisher: Humana New York, NY

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media, LLC, part of Springer Nature 2022

  • Hardcover ISBN: 978-1-0716-2151-6Published: 22 June 2022

  • Softcover ISBN: 978-1-0716-2154-7Published: 23 June 2023

  • eBook ISBN: 978-1-0716-2152-3Published: 21 June 2022

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XI, 278

  • Number of Illustrations: 5 b/w illustrations, 58 illustrations in colour

  • Topics: Biotechnology, Human Genetics

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