MoCloFlex: A Modular Yet Flexible Cloning System
MoCloFlex: A Modular Yet Flexible Cloning System
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DOI：
10.3389/fbioe.2019.00271
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发表时间：
2019-10-17
影响因子：
5.7
通讯作者：
Sobetzko, Patrick
中科院分区：
文献类型：
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作者：
Klein, Carlo A.;Emde, Leonie;Sobetzko, Patrick
现代克隆技术正在逐步取代传统的克隆方法。目前的系统利用具有通过金门反应连接的预定义DNA部分的文库。然而，这些系统仍然存在特定的兼容性和缺乏相互兼容性的问题。在这里，我们提出了灵活的模块化克隆（MoCloFlex），它通过引入一组允许自由单元排列的连接器和位置向量来克服这种不确定性。因此，我们的系统提供了一种方便的方法来设计和构建定制质粒，以及大型构建体的迭代组装。为了支持合成生物学的标准化，MoCloFlex与成熟的模块化克隆标准兼容。在这里，我们提出并表征MoCloFlex的各种应用程序，在一个单一的限制性连接反应多达12个片段。
Modern cloning solutions are gradually replacing classical cloning methods. Current systems make use of libraries with predefined DNA parts that are joined by Golden-Gate reactions. However, these systems still suffer from specific inflexibilities and the lack of inter-compatibility. Here, we present Flexible Modular Cloning (MoCloFlex) which overcomes this inflexibility by introducing a set of linker- and position-vectors allowing free unit arrangement. Our system, therefore, provides a convenient way to design and build custom plasmids, and iterative assembly of large constructs. To support standardization in synthetic biology, MoCloFlex is compatible with the well-established Modular Cloning standard. Here, we present and characterize MoCloFlex for various applications with up to 12 fragments in a single restriction-ligation reaction.