A modular system for Clostridium shuttle plasmids
A modular system for Clostridium shuttle plasmids
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DOI：
10.1016/j.mimet.2009.05.004
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发表时间：
2009-07-01
影响因子：
2.2
通讯作者：
Minton, Nigel P.
中科院分区：
文献类型：
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作者：
Heap, John T.;Pennington, Oliver J.;Minton, Nigel P.
尽管它们在医学和工业上具有重要意义，但与芽孢杆菌属中的需氧细菌相比，我们对梭状芽胞杆菌属生物学的基本了解是基本的。一个主要的因素是梭状芽胞杆菌分子生物学家可用的基因工具相对缺乏复杂性，这些工具还不成熟，显然需要开发。重组、复制性质粒在梭状芽孢杆菌中的转移和维持已有报道，并且已知几种适合作为穿梭质粒组分的元件。然而，到目前为止，这些组分只在完全不同的质粒环境中可用，并且它们的用途还没有得到广泛的探索。在这里，我们描述了梭状芽胞杆菌-大肠杆菌穿梭质粒标准化模块系统的规范、设计和构建。将现有的复制子和可选择的标记与一种新的梭状芽胞杆菌复制子结合在一起。研究人员比较了这些组件的特性，这些数据使研究人员能够确定潜在适合特定主机和应用程序的组件组合。该系统已经在我们的实验室进行了广泛的测试，并在所有正在进行的重组工作中使用。我们建议，采用这种模块化系统作为标准，将对梭状芽胞杆菌研究社区有很大的好处，我们邀请他们使用该系统并为其做出贡献。(C)2009爱思唯尔B.V.保留所有权利。
Despite their medical and industrial importance, our basic understanding of the biology of the genus Clostridium is rudimentary in comparison to their aerobic counterparts in the genus Bacillus. A major contributing factor has been the comparative lack of sophistication in the gene tools available to the clostridial molecular biologist, which are immature, and in clear need of development. The transfer and maintenance of recombinant, replicative plasmids into various species of Clostridium has been reported, and several elements suitable as shuttle plasmid components are known. However, these components have to-date only been available in disparate plasmid contexts, and their use has not been broadly explored. Here we describe the specification, design and construction of a standardized modular system for Clostridium-Escherichia coli shuttle plasmids. Existing replicons and selectable markers were incorporated, along with a novel clostridial replicon. The properties of these components were compared, and the data allow researchers to identify combinations of components potentially suitable for particular hosts and applications. The system has been extensively tested in our laboratory, where it is utilized in all ongoing recombinant work. We propose that adoption of this modular system as a standard would be of substantial benefit to the Clostridium research community, whom we invite to use and contribute to the system. (C) 2009 Elsevier B.V. All rights reserved.