利用 PEDA 方法进行体内 DNA 组装。
In Vivo DNA Assembly Using the PEDA Method.
机构信息
State Key Laboratory of Microbial Technology, Shandong University, Qingdao, People's Republic of China.
Shandong Lishan Biotechnology Co. LTD, Jinan, People's Republic of China.
出版信息
Methods Mol Biol. 2024;2760:437-445. doi: 10.1007/978-1-0716-3658-9_24.
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.
简单高效的 DNA 组装方法已在合成生物学中得到广泛应用。本文提供了一种新的 PEDA（噬菌体酶辅助体内 DNA 组装）方法，用于在多种微生物中直接进行单个 DNA 片段的体内组装，这些微生物包括大肠杆菌、氧化亚铁硫杆菌、恶臭假单胞菌、植物乳杆菌和解脂耶氏酵母。PEDA 允许具有 5bp 同源序列的 DNA 片段进行体内组装，其效率与传统的体外 DNA 组装相当，这将极大地推动合成生物学的快速发展。
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