Standardized Modular Assembly of Polycistronic Operons with Modular Cloning (MoClo) using the <em>In-Cloning</em> toolkit

Jelmar de Vries; Timon A. Lindeboom; Stijn T. de Vries; Daniel Schindler; Stefan A. Hoffmann

doi:10.3791/68103

使用 In-Cloning 工具包对具有模块化克隆的多顺反子纵子（MoClo）进行标准化模块化组装

JoVE Journal
Bioengineering

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JoVE Journal Bioengineering

Standardized Modular Assembly of Polycistronic Operons with Modular Cloning (MoClo) using the In-Cloning toolkit

使用 In-Cloning 工具包对具有模块化克隆的多顺反子纵子（MoClo）进行标准化模块化组装

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06:28 min

September 2, 2025

DOI: 10.3791/68103-v

Jelmar de Vries¹, Timon A. Lindeboom², Stijn T. de Vries², Daniel Schindler^2,3,4, Stefan A. Hoffmann¹

¹Laboratory of Systems and Synthetic Biology (SSB),Wageningen University and Research, Agrotechnology and Food Sciences, ²Max Planck Institute for Terrestrial Microbiology, ³Center for Molecular Biology of Heidelberg University (ZMBH), ⁴Center for Synthetic Microbiology,Philipps-University Marburg

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本文提出了一个分步协议，演示了模块化克隆（MoClo）如何适用于多顺反子纵子的克隆。

我开发了能够从小分子片段到染色体大小构造体模块化组装DNA的方法，并用这些人工染色体重新编程细胞。该领域的一项重要创新是开发了模块化克隆工具包，使得能够从标准化分子元件库中模块化组装多基因构造体。模块化克隆工具包通常设计用于克隆单顺电子转录单元。

该方案展示了一种可推广的多顺子转录单元组装方法，结合克隆内工具包。这种克隆多变电子转录单元的策略使编码序列能够灵活地用于单个转录单元或多变电子排列。开始时，选择准备组装成转录单元的0级部件。

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