Nucleic Acids Research Advance Access published online on October 24, 2007
Nucleic Acids Research, doi:10.1093/nar/gkm818
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Methods Online |
A comprehensive set of plasmids for vanillate- and xylose-inducible gene expression in Caulobacter crescentus
1Max Planck Institute for Terrestrial Microbiology, Karl-von-Frisch-Straße, 35043 Marburg, Germany and 2Department of Developmental Biology, Stanford University School of Medicine, Beckman Center B300, 279 Campus Drive, Stanford, CA 94305, USA
*To whom correspondence should be addressed. Tel: +49 6421 178330, Fax: +49 6421 178209; Email: thanbichler{at}mpi-marburg.mpg.de
Received August 24, 2007. Revised September 17, 2007. Accepted September 18, 2007.
Caulobacter crescentus is widely used as a powerful model system for the study of prokaryotic cell biology and development. Analysis of this organism is complicated by a limited selection of tools for genetic manipulation and inducible gene expression. This study reports the identification and functional characterization of a vanillate-regulated promoter (Pvan) which meets all requirements for application as a multi-purpose expression system in Caulobacter, thus complementing the established xylose-inducible system (Pxyl). Furthermore, we introduce a newly constructed set of integrating and replicating shuttle vectors that considerably facilitate cell biological and physiological studies in Caulobacter. Based on different narrow and broad-host range replicons, they offer a wide choice of promoters, resistance genes, and fusion partners for the construction of fluorescently or affinity-tagged proteins. Since many of these constructs are also suitable for use in other bacteria, this work provides a comprehensive collection of tools that will enrich many areas of microbiological research.
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