Part:BBa_K2715043
Alternative sigma factor TcdR regulating toxin expression in C. difficile
Usage and Biology
This biobrick codes for the TcdR regulatory protein which regulates toxin A expression in Clostridium difficile. It is an alternative sigma factor and acts as an activator to positively regulate the activity of the tcdA promoter region. It was important when performing our assay to investigate the effect of CRISPRi methods on the tcdA promoter region to include this regulatory protein, as it is a key regulator.
Characterisation
This part was characterised as part of the composite BBa_K2715044, which was provided in addition to the composite parts BBa_K2715005, BBa_K2715006, BBa_K2715038, BBa_K2715039, BBa_K2715040, BBa_K2715041 inside an E. coli strain, each on a separate plasmid. In conjunction this enabled a GusA assay to give an indication of the level of CRISPRi interference of each of the composite parts, which gave a good representation of the efficiency of each synthetic guide. The full characterisation of each of the guides can be found through their links above.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 259
Illegal XbaI site found at 475 - 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 259
Illegal XbaI site found at 475 - 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 259
Illegal XbaI site found at 475 - 1000COMPATIBLE WITH RFC[1000]
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