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Team:Bielefeld-CeBiTec/Project/Biosafety
From 2014.igem.org
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| - | + | We work on two different biosafety systems.<br> | |
| + | First of all we aim to implement an selection system without antibiotics. The target of this system is to perform two deletions in the KRX genome for two genes encoding the D-alanin racemase. The D-alanin racemase catalyses the reaction from L-alanin to D-alanin. D-alanin is an important factor for the bacterial cell wall. Every plasmid which is used for the transformation should carry the D-alanin racemase to complement the deletion. Without the uptake of the plasmid the bacterial cells will die because of their lack of cell wall production.<br> | ||
| + | For second system rhamnose is essential for the media. In the presence of rhamnose a rhamnose dependent promotor is activated which lead to the expression of araC and the D-alanin racemase. The protein of araC inhibits the expression of the pBAD promotor. In the absence of rhamnose two kill switches are activated. If the rhamnose dependent promotor is not activated D-alanin could not be produced which lead to the lysis of the cell wall. In addition araC is not expressed anymore. This leads to the expression of the RNase Ba (Barnase) which leads to the lysis of intracellular RNA.<br><br> | ||
| + | <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Results/Biosafety"> Here</a> you will find the results of our biosafety. | ||
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| - | Instruction manual | + | Instruction manual GenTSV (S1) <a href = "https://static.igem.org/mediawiki/2014/e/ec/Bielefeld-CeBiTec_2014-07-20_2014-GenTSV-S1-CeBiTec.pdf" target="_blank"> <img src="https://static.igem.org/mediawiki/2014/b/bd/Bielefeld-CeBiTec_2014-08-12_pdf_Icon.png" height="30px"> </a> |
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| - | + | Instruction manual GefStoffV <a href = "https://static.igem.org/mediawiki/2014/d/d6/Bielefeld-CeBiTec_2014-07-20_2014-GefStoffV-CeBiTec.pdf" target="_blank"> <img src="https://static.igem.org/mediawiki/2014/b/bd/Bielefeld-CeBiTec_2014-08-12_pdf_Icon.png" height="30px"> </a> | |
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Revision as of 19:52, 19 August 2014
Biosafety
We work on two different biosafety systems.
First of all we aim to implement an selection system without antibiotics. The target of this system is to perform two deletions in the KRX genome for two genes encoding the D-alanin racemase. The D-alanin racemase catalyses the reaction from L-alanin to D-alanin. D-alanin is an important factor for the bacterial cell wall. Every plasmid which is used for the transformation should carry the D-alanin racemase to complement the deletion. Without the uptake of the plasmid the bacterial cells will die because of their lack of cell wall production.
For second system rhamnose is essential for the media. In the presence of rhamnose a rhamnose dependent promotor is activated which lead to the expression of araC and the D-alanin racemase. The protein of araC inhibits the expression of the pBAD promotor. In the absence of rhamnose two kill switches are activated. If the rhamnose dependent promotor is not activated D-alanin could not be produced which lead to the lysis of the cell wall. In addition araC is not expressed anymore. This leads to the expression of the RNase Ba (Barnase) which leads to the lysis of intracellular RNA.
Here you will find the results of our biosafety.
First of all we aim to implement an selection system without antibiotics. The target of this system is to perform two deletions in the KRX genome for two genes encoding the D-alanin racemase. The D-alanin racemase catalyses the reaction from L-alanin to D-alanin. D-alanin is an important factor for the bacterial cell wall. Every plasmid which is used for the transformation should carry the D-alanin racemase to complement the deletion. Without the uptake of the plasmid the bacterial cells will die because of their lack of cell wall production.
For second system rhamnose is essential for the media. In the presence of rhamnose a rhamnose dependent promotor is activated which lead to the expression of araC and the D-alanin racemase. The protein of araC inhibits the expression of the pBAD promotor. In the absence of rhamnose two kill switches are activated. If the rhamnose dependent promotor is not activated D-alanin could not be produced which lead to the lysis of the cell wall. In addition araC is not expressed anymore. This leads to the expression of the RNase Ba (Barnase) which leads to the lysis of intracellular RNA.
Here you will find the results of our biosafety.
Instruction manual GenTSV (S1) 
Instruction manual GefStoffV
