pBaSysBioll: an integrative plasmid generating gfp transcriptional fusions for high-throughput analysis of gene expression in Bacillus subtilis

被引：50

作者：

Botella, Eric ^{[1
]}

Fogg, Mark ^{[2
]}

Jules, Matthieu ^{[3
]}

Piersma, Sjouke

Doherty, Geoff ^{[6
]}

Hansen, Annette ^{[1
]}

Denham, Emma. L. ^{[4
,5
]}

Le Chat, Ludovic ^{[3
]}

Veiga, Patrick ^{[3
]}

Bailey, Kirra ^{[6
]}

Lewis, Peter J. ^{[6
]}

van Dijl, Jan Maarten ^{[4
]}

Aymerich, Stephane ^{[3
]}

Wilkinson, Anthony J. ^{[2
]}

Devine, Kevin M. ^{[1
]}

机构：

[1] Trinity Coll Dublin, Smurfit Inst Genet, Dublin 2, Ireland

[2] Univ York, Dept Chem, York Struct Biol Lab, York YO10 5YW, N Yorkshire, England

[3] INRA & AgroParisTech, Microbiol & Genet Mol, F-78850 Thiverval Grignon, France

[4] Univ Med Ctr Groningen, Dept Med Microbiol, Groningen, Netherlands

[5] Univ Groningen, Groningen, Netherlands

[6] Univ Newcastle, Sch Environm & Life Sci, Callaghan, NSW 2308, Australia

来源：

MICROBIOLOGY-SGM | 2010年 / 156卷

关键词：

LIGATION-INDEPENDENT CLONING; PHOPR 2-COMPONENT SYSTEMS; GREEN FLUORESCENT PROTEIN; GLUCOSE; TARGET; YYCFG;

D O I：

10.1099/mic.0.035758-0

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

Plasmid pBaSysBioll was constructed for high-throughput analysis of gene expression in Bacillus subtilis. It is an integrative plasmid with a ligation-independent cloning (LIC) site, allowing the generation of transcriptional gfpmut3 fusions with desired promoters. Integration is by a Campbell-type event and is non-mutagenic, placing the fusion at the homologous chromosomal locus. Using phoA, murAA, gapB, ptsG and cggR promoters that are responsive to phosphate availability, growth rate and carbon source, we show that detailed profiles of promoter activity can be established, with responses to changing conditions being measurable within 1 min of the stimulus. This makes pBaSysBioll a highly versatile tool for real-time gene expression analysis in growing cells of B. subtilis.

引用

页码：1600 / 1608

页数：9

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