Mini-Tn7 Insertion in an Artificial attTn7 Site Enables Depletion of the Essential Master Regulator CtrA in the Phytopathogen Agrobacterium tumefaciens
Mini-Tn7 Insertion in an Artificial attTn7 Site Enables Depletion of the Essential Master Regulator CtrA in the Phytopathogen Agrobacterium tumefaciens
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DOI：
10.1128/aem.01392-16
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发表时间：
2016-08-01
影响因子：
4.4
通讯作者：
Brown, Pamela J. B.
中科院分区：
文献类型：
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作者：
Figueroa-Cuilan, Wanda;Daniel, Jeremy J.;Brown, Pamela J. B.
农杆菌中许多过程的机制研究一直受到缺乏遗传工具来表征必需基因的阻碍。在这项研究中，我们使用了一种基于Tn7的方法，从染色体上的一个工程化位点诱导控制转录。我们证明，这种方法能够更严格地控制诱导型启动子比质粒为基础的系统，并可用于耗尽研究。该方法能够构建缺失菌株，以表征A.根瘤菌在这里，我们使用的战略，以耗尽的alphaproteobacterial主调节CtrA，并发现，这种必需的基因的耗尽导致戏剧性的圆的细胞，这成为nonviolet.IMPORTANCEAgrobacterium根癌农杆菌是一种细菌植物病原体和天然的遗传工程师。因此，对细胞周期进程、DNA复制和分离、细胞生长和分裂等基本过程的研究可能会为限制疾病或改善生物技术应用提供见解。
Mechanistic studies of many processes in Agrobacterium tumefaciens have been hampered by a lack of genetic tools for characterization of essential genes. In this study, we used a Tn7-based method for inducible control of transcription from an engineered site on the chromosome. We demonstrate that this method enables tighter control of inducible promoters than plasmid-based systems and can be used for depletion studies. The method enables the construction of depletion strains to characterize the roles of essential genes in A. tumefaciens. Here, we used the strategy to deplete the alphaproteobacterial master regulator CtrA and found that depletion of this essential gene results in dramatic rounding of cells, which become nonviable.IMPORTANCEAgrobacterium tumefaciens is a bacterial plant pathogen and natural genetic engineer. Thus, studies of essential processes, including cell cycle progression, DNA replication and segregation, cell growth, and division, may provide insights for limiting disease or improving biotechnology applications.