Team:Bielefeld-Germany/Biosafety/Biosafety System M

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==<span style = "color:#f88101; padding-left:22px">'''Safety system M (medium)'''</span> ==
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<h1>Biosafety System TetOR alive</h1>
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<a href="https://2013.igem.org/Team:Bielefeld-Germany/Project/Abstract">Projects Overview</a></div>
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<a href="https://2013.igem.org/Team:Bielefeld-Germany/Project/Nanowires#Theory">Theory</a></div>
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<a href="https://2013.igem.org/Team:Bielefeld-Germany/Project/Nanowires#Genetic_Approach">Genetic Approach</a></div>
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<a href="https://2013.igem.org/Team:Bielefeld-Germany/Project/Nanowires#Results">Results</a></div>
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==Overview==
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[[Image:Bielefeld-germany-project-overview-nanowires.png|left|thumb|250px|'''Figure 1:''' Principle of electron transfer from bacteria to anode via nanowires.]]
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The tetracyclin repressor (TetR)/ operator (TetO) originally is used by E. coli to work against the antibiotic tetracycline but in many cases it is used for regulated expression for industrial processes. When there is no tetracycline available the TetR binds with high affinity the tetracycline operator. When tetracycline is available the TetR switches his conformation and so it comes to a dissolution of the TetR and the TetO. Because of this the polymerase isn’t enhanced anymore and is able to express the genes which lies behind the TetO. In our system the TetR is under the control of a rhamnose promotor (rha-promotor) which only works in the presence of rhamnose. When the bacteria would break out of the MFC there wouldn’t be enough rhamnose in the environment to activate the promotor in a way that enough TetR would be produced to block the polymerase by binding at the TetO. Therefore the polymerase binds to the promotor of TetO and it comes to the expression of RNase Ba and the degradation of the DNA.  
The tetracyclin repressor (TetR)/ operator (TetO) originally is used by E. coli to work against the antibiotic tetracycline but in many cases it is used for regulated expression for industrial processes. When there is no tetracycline available the TetR binds with high affinity the tetracycline operator. When tetracycline is available the TetR switches his conformation and so it comes to a dissolution of the TetR and the TetO. Because of this the polymerase isn’t enhanced anymore and is able to express the genes which lies behind the TetO. In our system the TetR is under the control of a rhamnose promotor (rha-promotor) which only works in the presence of rhamnose. When the bacteria would break out of the MFC there wouldn’t be enough rhamnose in the environment to activate the promotor in a way that enough TetR would be produced to block the polymerase by binding at the TetO. Therefore the polymerase binds to the promotor of TetO and it comes to the expression of RNase Ba and the degradation of the DNA.  
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==Genetic Approach==
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[[File:IGEM Bielefeld 2013 Biosafety System M.png|600px|thumb|center|System M in the MFC: In this case the mikroorganism is in the MFC with sufficient L-rhamnose. It comes to an expression of TetR which blocks TetO by binding and alr which switches L-alanine to D-alanine. Because of the fact that TetR blocks TetO the RNase Ba can't expressed.]]
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[[File:IGEM Bielefeld 2013 Biosafety System M ohne Rhamnose.png|600px|thumb|center|System M outside of the MFC: In this case the mikroorganism could get out of the MFC by damage or incorrect handling. Outside of the MFC there isn't enough L-rhamnose. So TetR doesn't block TetO anymore so the degradation process is induced by activating the TetO and the ensuing expression of RNase Ba. E.coli dies.]]
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[[File:IGEM Bielefeld 2013 biosafety TetR.png]]
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[[File:IGEM Bielefeld 2013 biosafety alr.png]]
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[[File:IGEM Bielefeld 2013 biosafety Terminator.png]]
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[[File:IGEM Bielefeld 2013 biosafety TetO.png]]
[[File:IGEM Bielefeld 2013 biosafety TetO.png]]
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[[File:IGEM Bielefeld 2013 biosafety RNase Ba.png]]
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==Results==
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[[File:IGEM Bielefeld 2013 Biosafety System M.png|600px|thumb|center|System M in the MFC: In this case the mikroorganism is in the MFC with sufficient L-rhamnose. It comes to an expression of TetR which blocks TetO by binding and alr which switches L-alanine to D-alanine. Because of the fact that TetR blocks TetO the RNase Ba can't expressed.]]
 
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==References==
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[[File:IGEM Bielefeld 2013 Biosafety System M ohne Rhamnose.png|600px|thumb|center|System M outside of the MFC: In this case the mikroorganism could get out of the MFC by damage or incorrect handling. Outside of the MFC there isn't enough L-rhamnose. So TetR doesn't block TetO anymore so the degradation process is induced by activating the TetO and the ensuing expression of RNase Ba. E.coli dies.]]
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Revision as of 22:28, 29 September 2013



Biosafety System TetOR alive


Overview

Figure 1: Principle of electron transfer from bacteria to anode via nanowires.

The tetracyclin repressor (TetR)/ operator (TetO) originally is used by E. coli to work against the antibiotic tetracycline but in many cases it is used for regulated expression for industrial processes. When there is no tetracycline available the TetR binds with high affinity the tetracycline operator. When tetracycline is available the TetR switches his conformation and so it comes to a dissolution of the TetR and the TetO. Because of this the polymerase isn’t enhanced anymore and is able to express the genes which lies behind the TetO. In our system the TetR is under the control of a rhamnose promotor (rha-promotor) which only works in the presence of rhamnose. When the bacteria would break out of the MFC there wouldn’t be enough rhamnose in the environment to activate the promotor in a way that enough TetR would be produced to block the polymerase by binding at the TetO. Therefore the polymerase binds to the promotor of TetO and it comes to the expression of RNase Ba and the degradation of the DNA.







Genetic Approach

System M in the MFC: In this case the mikroorganism is in the MFC with sufficient L-rhamnose. It comes to an expression of TetR which blocks TetO by binding and alr which switches L-alanine to D-alanine. Because of the fact that TetR blocks TetO the RNase Ba can't expressed.



System M outside of the MFC: In this case the mikroorganism could get out of the MFC by damage or incorrect handling. Outside of the MFC there isn't enough L-rhamnose. So TetR doesn't block TetO anymore so the degradation process is induced by activating the TetO and the ensuing expression of RNase Ba. E.coli dies.


IGEM Bielefeld 2013 biosafety Rhamnose-promoter.png



IGEM Bielefeld 2013 biosafety TetR.png


IGEM Bielefeld 2013 biosafety alr.png


IGEM Bielefeld 2013 biosafety Terminator.png


IGEM Bielefeld 2013 biosafety TetO.png


IGEM Bielefeld 2013 biosafety RNase Ba.png


Results

References







Contents