RT Journal Article SR Electronic(1) A1 Botella, Eric A1 Fogg, Mark A1 Jules, Matthieu A1 Piersma, Sjouke A1 Doherty, Geoff A1 Hansen, Annette A1 Denham, Emma. L. A1 Le Chat, Ludovic A1 Veiga, Patrick A1 Bailey, Kirra A1 Lewis, Peter J. A1 van Dijl, Jan Maarten A1 Aymerich, Stéphane A1 Wilkinson, Anthony J. A1 Devine, Kevin M.YR 2010 T1 pBaSysBioII: an integrative plasmid generating gfp transcriptional fusions for high-throughput analysis of gene expression in Bacillus subtilis JF Microbiology, VO 156 IS 6 SP 1600 OP 1608 DO https://doi.org/10.1099/mic.0.035758-0 PB Microbiology Society, SN 1465-2080, AB 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. , UL https://www.microbiologyresearch.org/content/journal/micro/10.1099/mic.0.035758-0