Difference between revisions of "Part:BBa K2992001"
(→‎Characterisation)
Line 11: Line 11:
 
===Characterisation===
 
===Characterisation===
  
−
This basic part was characterised as part of our composite parts 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 )]. This part was used as a promoter which functions in both the Gram-negative E. coli and the Gram-positive Clostridium <i>sporogenens</i>. More information can be found on our [https://2019.igem.org/Team:Nottingham/Results Results Page].<br> <br> <br>
+
This basic part was used for the assembly of our composite parts and characterised using 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 )]. This part was used as a promoter which functions in both the Gram-negative E. coli and the Gram-positive Clostridium <i>sporogenens</i>. More information can be found on our [https://2019.igem.org/Team:Nottingham/Results Results Page].<br> <br> <br>
 
<br>
 
<br>
 
https://2019.igem.org/wiki/images/5/5e/T--Nottingham--Basic4.png
 
https://2019.igem.org/wiki/images/5/5e/T--Nottingham--Basic4.png

Revision as of 18:16, 21 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 of the toxin regulator botR and our chosen reporter genes of interest which have been placed under the control of the neurotoxin and neurotoxin-associated promoters. Doing so allows us to link volatile reporter production with neurotoxin production following food manufacturing processes.

Characterisation

This basic part was used for the assembly of our composite parts and characterised using using 3 difference reporters: GusA (BBa_K2992039), FAST (BBa_K2992044) and acetone (BBa_K2992036 ). This part was used as a promoter which functions in both the Gram-negative E. coli and the Gram-positive Clostridium sporogenens. More information can be found on our Results Page.



T--Nottingham--Basic4.png
T--Nottingham--Basic3.png
Characterisation of this promoter against Pfdx, Pthl and PbotR using FAST fluorescent assay, showed Pntnh to be an effective promoter in E.coli and only slightly stronger than no promoter in C.sporogenes.


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.