Difference between revisions of "Part:BBa K823048"

 
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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/Part:BBa_K823043 ECF41],[https://parts.igem.org/wiki/index.php?title=Part:BBa_K823041 PydfG] and [https://parts.igem.org/wiki/index.php?title=Part:BBa_K823044 MazF] in the <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].
  
Sigma 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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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.
  
 
<b>This Platereader measurment is the first and not reproduced to this point!</b>
 
<b>This Platereader measurment is the first and not reproduced to this point!</b>
 
[[Image:PlatereaderPspoIVBecf41PydfG.jpg|600px]]
 
[[Image:PlatereaderPspoIVBecf41PydfG.jpg|600px]]
  
This Platereader assay was done with the ''Bacillus subtilis'' strain W168 containing thrC::PspoIVB-ECF41 (through transformation with pSBBs4S-PspoIVB-ECF41) and sacA::pydfG-luxABCDE (through transformation with pSBBs3C-luxABCDE-PydfG). It shows that PspoIVB is activated first at six hours and then again starting at 18 hours. This is not as expected, as the peak at six hours is too early and probably not Sporulation specific. But it is an active promoter and seems to be activated during Sporulation.
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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>). It shows that P<sub>''spoIVB''</sub> is activated first at six hours and then again starting at 18 hours. This is not as expected, as the peak at six hours is too early and probably not Sporulation specific. But it is an active promoter and seems to be activated during Sporulation.
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 14:06, 26 September 2012

PspoIVB promoter regulated by sigma G

PspoIVB promoter is from B.subtilis and is a sporulation promoter activated late in sporulation by sigma G 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). It shows that PspoIVB is activated first at six hours and then again starting at 18 hours. This is not as expected, as the peak at six hours is too early and probably not Sporulation specific. But it is an active promoter and seems to be activated during Sporulation.

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]