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Synthetic Biology pp 205–212Cite as

Combinatorial Evolution of DNA with RECODE

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1772))

Abstract

In past decades, DNA engineering protocols have led to the rapid development of synthetic biology. To engineer the natural proteins, many directed evolution methods based on molecular biology have been presented for generating genetic diversity or obtaining specific properties. Here, we provide a simple (PCR operation), efficient (larger amount of products), and powerful (multiple point mutations, deletions, insertions, and combinatorial multipoint mutagenesis) RECODE method, which is capable of reediting the target DNA flexibly to restructure regulatory regions and remodel enzymes by using the combined function of the thermostable DNA polymerase and DNA ligase in one pot. RECODE is expected to be an applicable choice to create diverse mutant libraries for rapid evolution and optimization of enzymes and synthetic pathways.

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Acknowledgments

This work was financially supported by the National Natural Science Foundation of China (31670092), the Fundamental Research Funds for the Central Universities (JUSRP51707A), and Program for Changjiang Scholars and Innovative Research Team in University (No. IRT_15R26).

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Authors and Affiliations

  1. The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, China

    Zhen Kang, Wenwen Ding, Peng Jin, Guocheng Du & Jian Chen

  2. Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University, Wuxi, Jiangsu, China

    Zhen Kang, Guocheng Du & Jian Chen

  3. The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi, China

    Zhen Kang

Authors
  1. Zhen Kang
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  2. Wenwen Ding
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  3. Peng Jin
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  4. Guocheng Du
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  5. Jian Chen
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Corresponding author

Correspondence to Zhen Kang .

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Editors and Affiliations

  1. Agilent Technologies, Inc, La Jolla, California, USA

    Jeffrey Carl Braman

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Kang, Z., Ding, W., Jin, P., Du, G., Chen, J. (2018). Combinatorial Evolution of DNA with RECODE. In: Braman, J. (eds) Synthetic Biology. Methods in Molecular Biology, vol 1772. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7795-6_11

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  • DOI: https://doi.org/10.1007/978-1-4939-7795-6_11

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7794-9

  • Online ISBN: 978-1-4939-7795-6

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