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.
Botella
Eric
E
Smurfit Institute of Genetics, Trinity College Dublin, Dublin 2, Ireland.
Fogg
Mark
M
York Structural Biology Laboratory, Department of Chemistry, University of York, York YO10 5YW, UK.
Jules
Matthieu
M
INRA and AgroParisTech, Microbiologie et Génétique Moléculaire, F-78850 Thiverval-Grignon, France.
Piersma
Sjouke
S
Department of Medical Microbiology, University Medical Center Groningen and University of Groningen, Hanzeplein 1, 9700 RB Groningen, The Netherlands.
Doherty
Geoff
G
School of Environmental and Life Sciences, University of Newcastle, Callaghan, NSW 2308, Australia.
Hansen
Annette
A
Smurfit Institute of Genetics, Trinity College Dublin, Dublin 2, Ireland.
Denham
Emma L
EL
Department of Medical Microbiology, University Medical Center Groningen and University of Groningen, Hanzeplein 1, 9700 RB Groningen, The Netherlands.
Le Chat
Ludovic
L
INRA and AgroParisTech, Microbiologie et Génétique Moléculaire, F-78850 Thiverval-Grignon, France.
Veiga
Patrick
P
INRA and AgroParisTech, Microbiologie et Génétique Moléculaire, F-78850 Thiverval-Grignon, France.
Bailey
Kirra
K
School of Environmental and Life Sciences, University of Newcastle, Callaghan, NSW 2308, Australia.
Lewis
Peter J
PJ
School of Environmental and Life Sciences, University of Newcastle, Callaghan, NSW 2308, Australia.
van Dijl
Jan Maarten
JM
Department of Medical Microbiology, University Medical Center Groningen and University of Groningen, Hanzeplein 1, 9700 RB Groningen, The Netherlands.
Aymerich
Stéphane
S
INRA and AgroParisTech, Microbiologie et Génétique Moléculaire, F-78850 Thiverval-Grignon, France.
Wilkinson
Anthony J
AJ
York Structural Biology Laboratory, Department of Chemistry, University of York, York YO10 5YW, UK.
Devine
Kevin M
KM
Smurfit Institute of Genetics, Trinity College Dublin, Dublin 2, Ireland.
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