Difference between revisions of "Part:BBa K823042"

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[[Image:SuicideSwitchgrafik.jpg|600px]]
 
[[Image:SuicideSwitchgrafik.jpg|600px]]
  
Our team used this part in the combination with [https://parts.igem.org/wiki/index.php?title=Part:BBa_K823048 P<sub>''spoIVB''</sub>]/[https://parts.igem.org/wiki/index.php?title=Part:BBa_K823042 P<sub>''sspK''</sub>],[https://parts.igem.org/Part:BBa_K823043 ecf41<sub>Bli aa  1-204</sub>] and [https://parts.igem.org/wiki/index.php?title=Part:BBa_K823044 MazF] in the [http://2012.igem.org/Team:LMU-Munich/Germination_Stop#Suicide_switch <b>Suicide</b> switch].
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Our team used this part in the combination with [https://parts.igem.org/Part:BBa_K823041 P<sub>''ydfG''</sub>],[https://parts.igem.org/Part:BBa_K823043 ecf41<sub>Bli aa  1-204</sub>] and [https://parts.igem.org/wiki/index.php?title=Part:BBa_K823044 MazF] in the [http://2012.igem.org/Team:LMU-Munich/Germination_Stop#Suicide_switch <b>Suicide</b> 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.
 
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.
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[[Image:PlatereaderPspoIVBecf41PydfG.jpg|600px]]
 
[[Image:PlatereaderPspoIVBecf41PydfG.jpg|600px]]
  
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>).
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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/Part:BBa_K823041 P<sub>''ydfG''</sub>].
  
  

Revision as of 13:57, 26 September 2012

PsspK promoter regulated by sigma G

A B. subtilis promoter regulated by sigma G late in Sporulation in the forespore

SuicideSwitchgrafik.jpg

Our team used this part in the combination with PydfG,ecf41Bli aa 1-204 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.


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]