Genome engineering of Agrobacterium tumefaciens using the lambda Red recombination system
Genome engineering of Agrobacterium tumefaciens using the lambda Red recombination system
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使用 lambda Red 重组系统对根癌农杆菌进行基因组工程
DOI：
10.1007/s00253-013-5412-x
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发表时间：
2014-03-01
影响因子：
5
通讯作者：
Zhang, Youming
中科院分区：
文献类型：
--
作者：
Hu, Shengbiao;Fu, Jun;Zhang, Youming
根癌农杆菌已被广泛用作植物转基因的工具。其基因组序列的可获得性应有助于定向工程改良的特性；然而，目前的基因组工程选择是费力的。在这里，我们研究了Lambda Red操纵子是否可以用于根癌农杆菌基因组的重组工程。首先，我们构建了一个利用四环素诱导启动子调控Red操纵子的根癌农杆菌表达载体。然后在根癌农杆菌中对这个多拷贝质粒进行自身修饰，以验证Red操纵子是有功能的。然后，我们对内源根癌农杆菌的致瘤质粒和线形染色体进行了改造。这些结果将重组工程技术扩展到新的宿主，并表明了一种快速、方便地设计根癌农杆菌基因组以进行功能基因组学和菌株改良的方法。
Agrobacterium tumefaciens has been widely used as a tool for transgenesis in plants. The availability of its genome sequence should facilitate the directed engineering of improved properties; however, the current genome engineering options are laborious. Here, we investigated whether the lambda Red operon can be applied for recombineering of the A. tumefaciens genome. First, we built an expression plasmid for A. tumefaciens employing a tetracycline-inducible promoter to regulate the Red operon. This multicopy plasmid was then itself modified in A. tumefaciens to verify that the Red operon was functional. Then, we modified the endogenous A. tumefaciens tumor-inducing plasmid and the linear chromosome. These results extend recombineering technology to a new host and indicate a fast and convenient way to engineer the A. tumefaciens genome for functional genomics and strain improvements.