Difference between revisions of "Part:BBa K2992001"
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<partinfo>BBa_K2992001 short</partinfo>
 
<partinfo>BBa_K2992001 short</partinfo>
  
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Regulator region comprising promoter 5'-UTR and RBS for the neurotoxin gene cluster in <i>Clostridum botulinum</i>, we will be using this promoter for the production of our reporter.
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Promoter region for the non-toxic non-haemagglutinin component of the neurotoxin gene cluster in <i>Clostridum botulinum</i>.  
  
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===Usage and Biology===
 
===Usage and Biology===
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P<i>ntnh</i> is found upstream of the 5'-UTR and RBS of the non-toxic non-haemagglutinin component (ntnh) of the neurotoxin gene cluster in <i>C. botulinum</i>. This promoter was used to drive expression of our chosen reporter genes of interest. Doing so, allowed us to link BotR expression with the transcriptional activation of our reporter genes.
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===Characterisation===
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This basic part was used for the assembly of our composite parts and characterised using 3 difference reporters: [https://parts.igem.org/Part:BBa_K2992039 GusA (BBa_K2992039)], [https://parts.igem.org/Part:BBa_K2992044 FAST (BBa_K2992044)] and [https://parts.igem.org/Part:BBa_K2992036 acetone (BBa_K2992036 )]. More information can be found on our [https://2019.igem.org/Team:Nottingham/Results Results Page].<br> <br> <br>
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https://2019.igem.org/wiki/images/5/5e/T--Nottingham--Basic4.png
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https://2019.igem.org/wiki/images/6/6b/T--Nottingham--Basic3.png
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Characterisation of this promoter (comprising parts [https://parts.igem.org/Part:BBa_K2992001 BBa_K2992001] and [https://parts.igem.org/Part:BBa_K2992015 BBa_K2992015]) against P<i>fdx</i>, P<i>thl</i> and P<i>botR</i> using FAST fluorescent assay, showed that P<i>ntnh</i> is a medium-strength promoter in E.<i>coli</i>, while being very weak in C.<i>sporogenes</i> (only slightly stronger than the promoterless control), as expected in the absence of BotR. <br>
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<partinfo>BBa_K2992001 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2992001 SequenceAndFeatures</partinfo>
  
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===References===
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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.
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Latest revision as of 02:55, 22 October 2019


PntnH

Promoter region for the non-toxic non-haemagglutinin component of the neurotoxin gene cluster in Clostridum botulinum.

Usage and Biology

Pntnh is found upstream of the 5'-UTR and RBS of the non-toxic non-haemagglutinin component (ntnh) of the neurotoxin gene cluster in C. botulinum. This promoter was used to drive expression of our chosen reporter genes of interest. Doing so, allowed us to link BotR expression with the transcriptional activation of our reporter genes.

Characterisation

This basic part was used for the assembly of our composite parts and characterised using 3 difference reporters: GusA (BBa_K2992039), FAST (BBa_K2992044) and acetone (BBa_K2992036 ). More information can be found on our Results Page.



T--Nottingham--Basic4.png
T--Nottingham--Basic3.png
Characterisation of this promoter (comprising parts BBa_K2992001 and BBa_K2992015) against Pfdx, Pthl and PbotR using FAST fluorescent assay, showed that Pntnh is a medium-strength promoter in E.coli, while being very weak in C.sporogenes (only slightly stronger than the promoterless control), as expected in the absence of BotR.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

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.