Difference between revisions of "Part:BBa K823043"
 
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This Platereader assay was done with the ''Bacillus subtilis'' strain W168 containing ''thrC''::P<sub>''spoIVB''</sub>-''ecf41<sub>Bli aa  1-204</sub>'' (through transformation with pSBBs4S-P<sub>''spoIVB''</sub>-''ecf41<sub>Bli aa  1-204</sub>'') and ''sacA''::P<sub>''ydfG''</sub>-''luxABCDE'' (through transformation with pSBBs3C-''luxABCDE''-P<sub>''ydfG''</sub>). That we get a signal shows that this is active together with [https://parts.igem.org/wiki/index.php?title=Part:BBa_K823041 P<sub>''ydfG''</sub>].
 
This Platereader assay was done with the ''Bacillus subtilis'' strain W168 containing ''thrC''::P<sub>''spoIVB''</sub>-''ecf41<sub>Bli aa  1-204</sub>'' (through transformation with pSBBs4S-P<sub>''spoIVB''</sub>-''ecf41<sub>Bli aa  1-204</sub>'') and ''sacA''::P<sub>''ydfG''</sub>-''luxABCDE'' (through transformation with pSBBs3C-''luxABCDE''-P<sub>''ydfG''</sub>). That we get a signal shows that this is active together with [https://parts.igem.org/wiki/index.php?title=Part:BBa_K823041 P<sub>''ydfG''</sub>].
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For the use of our BaKillus suicide switch the team 2014 of the LMU used the alternative sigma factor 2cf41
 
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===Usage and Biology===
 
  
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[[File:delay prezi.png|thumb|600px|center|Fig. 1. Platereader measurement of ECF41 delay]]
  
  

Latest revision as of 05:52, 1 November 2014

alternative sigma factor ecf41

This is an alternative sigma factor from B. lincheniformes regulating e.g. PydfG. It is the short version lacking the regulation domain and therefore constitutively on.

SuicideSwitchgrafik.jpg

Our team used this part in the combination with PspoIVB/PsspK,PydfG and MazF in the [http://2012.igem.org/Team:LMU-Munich/Germination_Stop#Suicide_switch Suicide switch].

σ G, the last sigma factor in the forespore, activates PspoIVB, producing ECF41 during Sporulation. This then activates the PydfG promoter producing MazF, theoretically killing cells after germination.

This Platereader measurment is the first and not reproduced to this point! PlatereaderPspoIVBecf41PydfG.jpg

This Platereader assay was done with the Bacillus subtilis strain W168 containing thrC::PspoIVB-ecf41Bli aa 1-204 (through transformation with pSBBs4S-PspoIVB-ecf41Bli aa 1-204) and sacA::PydfG-luxABCDE (through transformation with pSBBs3C-luxABCDE-PydfG). That we get a signal shows that this is active together with PydfG.


Fig. 1. Platereader measurement of ECF41 delay


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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 443
  • 1000
    COMPATIBLE WITH RFC[1000]