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首页> 外文期刊>Applied Microbiology >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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Mini-Tn7 Insertion in an Artificial attTn7 Site Enables Depletion of the Essential Master Regulator CtrA in the Phytopathogen Agrobacterium tumefaciens

机译：在人工attTn7站点中的Mini-Tn7插入使枯草根癌农杆菌中的基本主调节剂CtrA耗尽。

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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 Tn 7 -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.IMPORTANCE Agrobacterium 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.
机译：由于缺乏用于表征必需基因的遗传工具，根癌农杆菌中许多过程的机理研究受到了阻碍。在这项研究中，我们使用了基于Tn 7的方法来诱导控制染色体上工程位点的转录。我们证明，该方法比基于质粒的系统能够更严格地控​​制诱导型启动子，可用于耗竭研究。该方法使耗竭菌株的构建能够表征根癌农杆菌中必需基因的作用。在这里，我们使用了该策略来耗尽α变形杆菌主调节剂CtrA，并发现该必需基因的耗尽会导致细胞急剧变圆，从而变得不可行。因此，对基本过程的研究，包括细胞周期进程，DNA复制和分离，细胞生长和分裂，可能为限制疾病或改善生物技术应用提供见识。

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