pBaSysBioII: an integrative plasmid generating gfp transcriptional fusions for high-throughput analysis of gene expression in Bacillus subtilis.
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.
Smurfit Institute of Genetics, Trinity College Dublin, Dublin 2, Ireland.
Eric
E
Botella
York Structural Biology Laboratory, Department of Chemistry, University of York, York YO10 5YW, UK.
Mark
M
Fogg
INRA and AgroParisTech, Microbiologie et Génétique Moléculaire, F-78850 Thiverval-Grignon, France.
Matthieu
M
Jules
Department of Medical Microbiology, University Medical Center Groningen and University of Groningen, Hanzeplein 1, 9700 RB Groningen, The Netherlands.
Sjouke
S
Piersma
School of Environmental and Life Sciences, University of Newcastle, Callaghan, NSW 2308, Australia.
Geoff
G
Doherty
Smurfit Institute of Genetics, Trinity College Dublin, Dublin 2, Ireland.
Annette
A
Hansen
Department of Medical Microbiology, University Medical Center Groningen and University of Groningen, Hanzeplein 1, 9700 RB Groningen, The Netherlands.
Emma L
EL
Denham
INRA and AgroParisTech, Microbiologie et Génétique Moléculaire, F-78850 Thiverval-Grignon, France.
Ludovic
L
Le Chat
INRA and AgroParisTech, Microbiologie et Génétique Moléculaire, F-78850 Thiverval-Grignon, France.
Patrick
P
Veiga
School of Environmental and Life Sciences, University of Newcastle, Callaghan, NSW 2308, Australia.
Kirra
K
Bailey
School of Environmental and Life Sciences, University of Newcastle, Callaghan, NSW 2308, Australia.
Peter J
PJ
Lewis
Department of Medical Microbiology, University Medical Center Groningen and University of Groningen, Hanzeplein 1, 9700 RB Groningen, The Netherlands.
Jan Maarten
JM
van Dijl
INRA and AgroParisTech, Microbiologie et Génétique Moléculaire, F-78850 Thiverval-Grignon, France.
Stéphane
S
Aymerich
York Structural Biology Laboratory, Department of Chemistry, University of York, York YO10 5YW, UK.
Anthony J
AJ
Wilkinson
Smurfit Institute of Genetics, Trinity College Dublin, Dublin 2, Ireland.
Kevin M
KM
Devine
D001412:Bacillus subtilis / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D002244:Carbon / Q000378:metabolism; D015870:Gene Expression; D015964:Gene Expression Regulation, Bacterial; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D010710:Phosphates / Q000378:metabolism; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic
D010710:Phosphates; D049452:Green Fluorescent Proteins; D002244:Carbon