Difference between revisions of "Part:BBa K2992012"
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===Usage and Biology=== | ===Usage and Biology=== | ||
− | This promoter region is found naturally upstream of the <i>botR</i> 5’-UTR in <i>C. botulinum</i>. BotR is an alternative sigma factor involved in the positive regulation of botulinum neurotoxin and associated genes. In our project, we use P<i>botR</i> to drive the expression of <i>botR</i> ( | + | This promoter region is found naturally upstream of the <i>botR</i> 5’-UTR in <i>C. botulinum</i> [https://parts.igem.org/Part:BBa_K2992014 BBa_K2992014]. BotR is an alternative sigma factor involved in the positive regulation of botulinum neurotoxin and associated genes. In our project, we use P<i>botR</i> to drive the expression of <i>botR</i> ([https://parts.igem.org/Part:BBa_K2992002 BBa_K2992002]) which in turn, regulates the production of our reporter genes which we have placed under the control of a BotR-activated promoter ([https://parts.igem.org/Part:BBa_K2992028 BBa_K2992028], [https://parts.igem.org/Part:BBa_K2992029 BBa_K2992029], [https://parts.igem.org/Part:BBa_K2992030 BBa_K2992030], [https://parts.igem.org/Part:BBa_K2992034 BBa_K2992034], [https://parts.igem.org/Part:BBa_K2992035 BBa_K2992035], [https://parts.igem.org/Part:BBa_K2992036 BBa_K2992036]). Doing so allowed us to link BotR expression with the trasncriptional activation of our reporter genes. <br><br> |
===Characterisation=== | ===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 [https://2019.igem.org/Team:Nottingham/Results Results Page].<br> <br> <br> |
− | < | + | <br> |
− | <partinfo> | + | https://2019.igem.org/wiki/images/5/5e/T--Nottingham--Basic4.png |
+ | <br> | ||
+ | https://2019.igem.org/wiki/images/6/6b/T--Nottingham--Basic3.png | ||
+ | <br> | ||
+ | Characterisation of P<i>botR</i> (comprising parts [https://parts.igem.org/Part:BBa_K2992012 BBa_K2992012] and [https://parts.igem.org/Part:BBa_K2992014 BBa_K2992014]) against Pfdx, Pthl and Pntnh using the FAST fluorescent assay, showed PbotR to be a relatively weak promoter in both <i>E.coli</i> and <i>C.sporogenes</i>. | ||
+ | |||
+ | <partinfo>BBa_K2992000 SequenceAndFeatures</partinfo> | ||
===References=== | ===References=== | ||
− | Raffestin | + | 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. | ||
<!-- Uncomment this to enable Functional Parameter display | <!-- Uncomment this to enable Functional Parameter display |
Latest revision as of 02:48, 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 PbotR (comprising parts BBa_K2992012 and BBa_K2992014) against Pfdx, Pthl and Pntnh using the FAST fluorescent assay, showed PbotR to be a relatively weak promoter in both E.coli and C.sporogenes.
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 155
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.