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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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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