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 Microbioljournal_title
Applied and environmental microbiologyauthors
Figueroa-Cuilan W,Daniel JJ,Howell M,Sulaiman A,Brown PJdoi
10.1128/AEM.01392-16subject
Has Abstractpub_date
2016-07-29 00:00:00pages
5015-25issue
16eissn
0099-2240issn
1098-5336pii
AEM.01392-16journal_volume
82pub_type
杂志文章abstract::Genetic optimizations to achieve high-level production of three different proteins of medical importance for humans, granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon alpha 2b (IFN-alpha2b), and single-chain antibody variable fragment (scFv-phOx), were investigated during high-cell-density cultivat...
journal_title:Applied and environmental microbiology
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journal_title:Applied and environmental microbiology
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journal_title:Applied and environmental microbiology
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journal_title:Applied and environmental microbiology
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
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更新日期:1996-05-01 00:00:00
abstract::A molecular method based on restriction fragment length polymorphism (RFLP) of PCR-amplified fragments of the 23S rRNA gene was designed to rapidly identify Listeria strains to the species level. Two fragments (S1, 460 bp, and S2, 890 bp) were amplified from boiled DNA. S2 was cut with the restriction enzymes XmnI or ...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
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