Brune, Iris; Werner, Hendrikje; Hüser, Andrea T.; Kalinowski, Jörn; Pühler, Alfred; Tauch, Andreas: The DtxR protein acting as dual transcriptional regulator directs a global regulatory network involved in iron metabolism of Corynebacterium glutamicum. In: BMC Genomics. Jg.7 H. 1. 2006
Inhalt
- Background
- Results
- Phenotypic characterization of the dtxR mutant strain C. glutamicum IB2103
- Genome-wide transcriptional profiling of the dtxR mutant C. glutamicum IB2103 in comparison to the wild-type strain ATCC 13032 by DNA microarray hybridization
- Computational identification of potential DtxR binding sites in the genome sequence of C. glutamicum ATCC 13032
- Verification of differential expression of genes with predicted DtxR binding sites by real-time RT-PCR experiments
- Binding of purified DtxR protein to the identified sequence motifs as analyzed by DNA band shift assays
- Localization of DtxR binding sites in mapped promoter regions of differentially expressed genes
- Bioinformatics characterization and functional dissection of the DtxR regulon of C. glutamicum
- Genome-wide identification of putative DtxR binding sites in corynebacterial genome sequences and comparative content analysis of the deduced DtxR regulons
- Discussion
- Conclusion
- Methods
- Bacterial strains and growth conditions
- DNA isolation, manipulation and transfer
- PCR techniques and dtxR mutant construction
- Total RNA preparation and DNA microarray hybridization
- Bioinformatics tools for DtxR binding site prediction
- Real-time reverse transcription (RT)-PCR
- Construction and purification of a His-tagged DtxR protein
- DNA band shift assays to characterize binding of the purified DtxR protein
- Identification of transcriptional start sites by the RACE method
- Authors' contributions
- Additional material
- Acknowledgements
- References