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

Eric Botella, Mark Fogg, Matthieu Jules, Sjouke Piersma, Geoff Doherty, Annette Hansen, Emma L Denham, Ludovic Le Chat, Patrick Veiga, Kirra Bailey, Peter J Lewis, Jan Maarten van Dijl, Stéphane Aymerich, Anthony J Wilkinson, Kevin M Devine

Research output: Contribution to journalArticlepeer-review

48 Citations (Scopus)

Abstract

Plasmid pBaSysBioII 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 pBaSysBioII a highly versatile tool for real-time gene expression analysis in growing cells of B. subtilis.

Original languageEnglish
Pages (from-to)1600-1608
Number of pages9
JournalMicrobiology
Volume156
Issue number6
Early online date1 Jun 2010
DOIs
Publication statusE-pub ahead of print - 1 Jun 2010

Keywords

  • Bacillus subtilis/genetics
  • Base Sequence
  • Carbon/metabolism
  • Gene Expression
  • Gene Expression Regulation, Bacterial
  • Green Fluorescent Proteins/genetics
  • Molecular Sequence Data
  • Phosphates/metabolism
  • Plasmids
  • Promoter Regions, Genetic
  • Transcription, Genetic

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