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

Microbiology (Reading). 2010 Jun;156(Pt 6):1600-1608. doi: 10.1099/mic.0.035758-0. Epub 2010 Feb 11.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

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

Substances

  • Phosphates
  • Green Fluorescent Proteins
  • Carbon

Associated data

  • GENBANK/GU126429