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

Abstract:

UNLABELLED: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. 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.

journal_name

Appl Environ Microbiol

authors

Figueroa-Cuilan W,Daniel JJ,Howell M,Sulaiman A,Brown PJ

doi

10.1128/AEM.01392-16

subject

Has Abstract

pub_date

2016-07-29 00:00:00

pages

5015-25

issue

16

eissn

0099-2240

issn

1098-5336

pii

AEM.01392-16

journal_volume

82

pub_type

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