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In Vivo DNA Assembly Using the PEDA Method

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Synthetic Biology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2760))

Abstract

Simple and efficient DNA assembly methods have been widely used in synthetic biology. Here, we provide the protocol for the recently developed PEDA (phage enzyme-assisted in vivo DNA assembly) method for direct in vivo assembly of individual DNA parts in multiple microorganisms, such as Escherichia coli, Ralstonia eutropha, Pseudomonas putida, Lactobacillus plantarum, and Yarrowia lipolytica. PEDA allows in vivo assembly of DNA fragments with homologous sequences as short as 5 bp, and the efficiency is comparable to the prevailing in vitro DNA assembly, which will broadly boost the rapid progress of synthetic biology.

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Acknowledgments

This work was supported by grants from the National Key R&D Program of China (2021YFC2100500), the National Natural Science Foundation of China (31730003, 32200081), and Shandong Provincial Natural Science Foundation (ZR2021QC021).

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Authors and Affiliations

  1. State Key Laboratory of Microbial Technology, Shandong University, Qingdao, People’s Republic of China

    Tianyuan Su & Qingsheng Qi

  2. Shandong Lishan Biotechnology Co. LTD, Jinan, People’s Republic of China

    Qingxiao Pang

Authors
  1. Tianyuan Su
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  2. Qingxiao Pang
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  3. Qingsheng Qi
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Corresponding author

Correspondence to Qingsheng Qi .

Editor information

Editors and Affiliations

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

    Jeffrey Carl Braman

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Su, T., Pang, Q., Qi, Q. (2024). In Vivo DNA Assembly Using the PEDA Method. In: Braman, J.C. (eds) Synthetic Biology. Methods in Molecular Biology, vol 2760. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3658-9_24

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  • DOI: https://doi.org/10.1007/978-1-0716-3658-9_24

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3657-2

  • Online ISBN: 978-1-0716-3658-9

  • eBook Packages: Springer Protocols

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