Team:Trieste/parts/3

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<h2>Looking forward</h2>
<h2>Looking forward</h2>
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As we test that a single copy of CYM R in the plasmid can repress strictly the T5 cumate operator the next step is two integrate in the genome of the bacteria first a single copy of CYM R then the double copy. If this integration is verified we can have together with the T5 operator and the toxin (both cloned in the same plasmid) a simple and efficient system to control bacteria proliferation and specially to avoid the horizontal transfer.
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The next step will be the integration of two copies of CymR in the bacterial genome in order to regulate the toxin genes carried by the plasmid.
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Revision as of 10:45, 26 September 2012

BBa_K875003

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Description

This composite produces constitutively the CymR regulator protein, that binds the Cumate Operator repressing the transcription from the promoter.


Assembly

Obtained by synthesis.


Results

CymR expression was confirmed by Western Blot analysis. Our CymR has a SV5 tag at the C-terminus in order to be detected by an anti-SV5 Ab. Its functionality was confirmed through the repression of T5 Cumate Operator-GFP.

The validation of the repression of the protein CymR on the T5 cumate operator was verified through the creation of a plasmid containing a single copy of CYM R and the T5 cumate operator upstream the GFP (I13504). We saw that the expression of the GFP was completely repressed in absence of p-cumate but it was active in presence of p-cumate.


Looking forward


The next step will be the integration of two copies of CymR in the bacterial genome in order to regulate the toxin genes carried by the plasmid.


Link to the Registry


Università degli studi di Trieste ICGEB Illy Fondazione Cassa di Risparmio
iGEM 2012 iGEM 2012 iGEM 2012 iGEM 2012 iGEM 2012 iGEM 2012
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