Mini-Tn7 Insertion in an Artificial attTn7 Site Enables Depletion of the Essential Master Regulator CtrA in the Phytopathogen Agrobacterium tumefaciens

作者：Figueroa Cuilan Wanda; Daniel Jeremy J; Howell Matthew; Sulaiman Aliyah; Brown Pamela J B

来源：Applied and Environmental Microbiology, 2016, 82(16): 5015-5025.

DOI：10.1128/AEM.01392-16

<jats:title>ABSTRACT</jats:title><jats:p>Mechanistic studies of many processes in<jats:named-content content-type="genus-species">Agrobacterium tumefaciens</jats:named-content>have been hampered by a lack of genetic tools for characterization of essential genes. In this study, we used a Tn<jats:italic>7</jats:italic>-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<jats:named-content content-type="genus-species">A. tumefaciens</jats:named-content>. 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.</jats:p><jats:p><jats:bold>IMPORTANCE</jats:bold><jats:named-content content-type="genus-species">Agrobacterium tumefaciens</jats:named-content>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.</jats:p>

• 出版日期2016-8