Difference between revisions of "Part:BBa K2992012"

Revision as of 02:02, 22 October 2019

PbotR from C. botulinum

Promoter region for botR in C. botulinum

Usage and Biology

This promoter region is found naturally upstream of the botR 5’-UTR in C. botulinum BBa_K2992014. BotR is an alternative sigma factor involved in the positive regulation of botulinum neurotoxin and associated genes. In our project, we use PbotR to drive the expression of botR (BBa_K2992002) which in turn, regulates the production of our reporter genes which we have placed under the control of a BotR-activated promoter (BBa_K2992028, BBa_K2992029, BBa_K2992030, BBa_K2992034, BBa_K2992035, BBa_K2992036). Doing so allowed us to link BotR expression with the trasncriptional activation of our reporter genes.

Characterisation

This basic part was used for the assembly of our composite parts and was characterised using FAST, GusA and acetone assays. More information can be found on our Results Page.





Characterisation of this promoter against Pfdx, Pthl and Pntnh using FAST fluorescent assay, showed PbotR to be a relatively weak promoter in both E.coli and C.sporogenes.

References

Raffestin, S., Dupuy, B., Marvaud, J. and Popoff, M. (2004). BotR/A and TetR are alternative RNA polymerase sigma factors controlling the expression of the neurotoxin and associated protein genes in Clostridium botulinum type A and Clostridium tetani. Molecular Microbiology, 55(1), pp.235-249.

Zhang, Z., Korkeala, H., Dahlsten, E., Sahala, E., Heap, J., Minton, N. and Lindström, M. (2013). Two-Component Signal Transduction System CBO0787/CBO0786 Represses Transcription from Botulinum Neurotoxin Promoters in Clostridium botulinum ATCC 3502. PLoS Pathogens, 9(3), p.e1003252.