Team:Michigan/Project
From 2009.igem.org
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- | ==<B><font size= | + | ==<B><font size=5>Overview</font></B>== |
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- | ==<B><font size= | + | ==<B><font size=5>Toluene Metabolism</font></B>== |
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- | ==<B><font size= | + | ==<B><font size=5>Suicide Mechanism</font></B>== |
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The Terminator's suicide mechanism, or kill switch, operates through the [http://partsregistry.org/wiki/index.php?title=Part:BBa_K112808<font color=navy><B>Enterobacteria phage T4 Lysis Device</B></font>] created by the Berkley 2008 team. We proposed two mechanisms for cell lysis: arabinose inducible suicide mechanism and suicide mechanism with tunable repression. These two kill switches could both appear, with others, in a final device, considering that severalfold kill switch redundancy will be necessary to prevent "runaway Terminators" (see Safety page). In the both models the Pu promoter is controlling the cell survival and we make the assumption that when the cells are used for bioremediation purposes they are placed in a toluene-contaminated environment. | The Terminator's suicide mechanism, or kill switch, operates through the [http://partsregistry.org/wiki/index.php?title=Part:BBa_K112808<font color=navy><B>Enterobacteria phage T4 Lysis Device</B></font>] created by the Berkley 2008 team. We proposed two mechanisms for cell lysis: arabinose inducible suicide mechanism and suicide mechanism with tunable repression. These two kill switches could both appear, with others, in a final device, considering that severalfold kill switch redundancy will be necessary to prevent "runaway Terminators" (see Safety page). In the both models the Pu promoter is controlling the cell survival and we make the assumption that when the cells are used for bioremediation purposes they are placed in a toluene-contaminated environment. | ||
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- | In this design, a repression mechanism is downstream of the Pu promoter, and this this represses the production of holin and lysozyme proteins. A constitutive promoter is placed in the front of the antiholin gene so antiholin is constantly produced. When toluene is present, it activates the Pu promoter which results in the repression of the promoter expressing holin and lysozyme. As a result, the cell survives. However, in the absence of toluene, the repressor protein is not produced, so holin and lysozyme levels rise to the point that the cells are lysed. | + | In this design, a repression mechanism is downstream of the Pu promoter, and this this represses the production of holin and lysozyme proteins. A constitutive promoter is placed in the front of the antiholin gene so antiholin is constantly produced. When toluene is present, it activates the Pu promoter which results in the repression of the promoter expressing holin and lysozyme. As a result, the cell survives. However, in the absence of toluene, the repressor protein is not produced, so holin and lysozyme levels rise to the point that the cells are lysed. This design is fairly tunable because we can choose which repression system to use, through which we can tweak the holin-antiholin balance. |
Revision as of 22:46, 21 October 2009
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