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Team:LMU-Munich/Data/Suicideswitch
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[[File:PlatereaderSuicideSwitch1.jpg|600px]] | [[File:PlatereaderSuicideSwitch1.jpg|600px]] | ||
| - | This module is the ''Bacillus subtilis'' strain containing thrC::PspoIVB-ECF41 (through transformation with pSB<sub>Bs</sub>4S-PspoIVB-ECF41) and sacA::pydfG-luxABCDE (through transformation with pSB<sub>Bs</sub>3C-''luxABCDE''-PydfG). <br> | + | This module is the ''Bacillus subtilis'' strain W168 containing thrC::PspoIVB-ECF41 (through transformation with pSB<sub>Bs</sub>4S-PspoIVB-ECF41) and sacA::pydfG-luxABCDE (through transformation with pSB<sub>Bs</sub>3C-''luxABCDE''-PydfG). <br> |
| - | This places the ''luxABCDE'' | + | This places the ''luxABCDE'' cassette in the place of the MazF later. So this output is the theoretical output of the MazF in our <b>Suicide</b> switch. <br> |
| + | Sigma G, the last sigma factor in the forespore, activates PspoIVB, producing ECF41. This then activates the PydfG promoter producing Lux ABCDE.<br> | ||
<b>This Platereader measurment is the first and not reproduced to this point!</b><br> | <b>This Platereader measurment is the first and not reproduced to this point!</b><br> | ||
In this platereader measurement we get a much too early peak of luminescence at about six hours, maybe due to unspecific small acitvation due to stress. As the ECF41 used is the short version which is constitutively on, this could lead to a high activation of the PydfG promoter. But there is also a significant activation at the point were sporulation should be in a late stage, though not finished.<br> | In this platereader measurement we get a much too early peak of luminescence at about six hours, maybe due to unspecific small acitvation due to stress. As the ECF41 used is the short version which is constitutively on, this could lead to a high activation of the PydfG promoter. But there is also a significant activation at the point were sporulation should be in a late stage, though not finished.<br> | ||
The early peak, if it is activated in all cells and not a subset of cells, will probably be sufficient to kill the vegetative cells too early.<br> | The early peak, if it is activated in all cells and not a subset of cells, will probably be sufficient to kill the vegetative cells too early.<br> | ||
| - | Though we will have to make this measurement reproducible, we are hoping that the other weaker promoter PsspK, which was not finished cloning, might be a better candidate. | + | Though we will have to make this measurement reproducible, we are hoping that the other, weaker promoter PsspK, which was not finished cloning, might be a better candidate. |
Revision as of 15:14, 25 September 2012
The LMU-Munich team is exuberantly happy about the great success at the World Championship Jamboree in Boston. Our project Beadzillus finished 4th and won the prize for the "Best Wiki" (with Slovenia) and "Best New Application Project".
[ more news ]
Platereader measurement of Suicide switch like module
This module is 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).
This places the luxABCDE cassette in the place of the MazF later. So this output is the theoretical output of the MazF in our Suicide switch.
Sigma G, the last sigma factor in the forespore, activates PspoIVB, producing ECF41. This then activates the PydfG promoter producing Lux ABCDE.
This Platereader measurment is the first and not reproduced to this point!
In this platereader measurement we get a much too early peak of luminescence at about six hours, maybe due to unspecific small acitvation due to stress. As the ECF41 used is the short version which is constitutively on, this could lead to a high activation of the PydfG promoter. But there is also a significant activation at the point were sporulation should be in a late stage, though not finished.
The early peak, if it is activated in all cells and not a subset of cells, will probably be sufficient to kill the vegetative cells too early.
Though we will have to make this measurement reproducible, we are hoping that the other, weaker promoter PsspK, which was not finished cloning, might be a better candidate.
