Team:INSA Toulouse/contenu/lab practice/results

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Revision as of 17:00, 2 October 2013

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Results

Characterizations

XOR1

In vitro

Description des conditions d'expérimentation



In vitro recombinase characterization protocol


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In vivo

Description des conditions d'expérimentation



In vitro recombinase characterization protocol


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AND1

In vitro

Description des conditions d'expérimentation



In vitro recombinase characterization protocol


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In vivo

Description des conditions d'expérimentation



In vitro recombinase characterization protocol


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Red Light Sensor

In vitro

Description des conditions d'expérimentation



In vitro recombinase characterization protocol


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In vivo

Description des conditions d'expérimentation



In vitro recombinase characterization protocol


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Blue Light Sensor

In vitro

Description des conditions d'expérimentation



In vitro recombinase characterization protocol


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In vivo

Description des conditions d'expérimentation



In vitro recombinase characterization protocol


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Pol T7

In vitro

Description des conditions d'expérimentation



In vitro recombinase characterization protocol


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In vivo

Description des conditions d'expérimentation



In vitro recombinase characterization protocol


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General inductor

In vivo

The couple TetR-pTet system construction is a first step to characterize the entire system. A constitutive promoter (BBa_J23116) was assemble to tetR and then assemble to pTet-rbs-RFP-term (Bba_I13521). pTet is a regulator constitutively ON that expresses the mRFP1 protein. TetR is an inducer that binds to pTet promoter and thus, represses expression the downstream system. Supply of tetracycline or its analog aTc (anhydrotetracycline) is known to bind to tetR and invert the operation (inhibits expression of mRFP in this case).

After 18 hours, clones containing this assembly (BBa_J23116-TetR-pTet-RFP) show a leaky basal expression of mRFP. We suppose that the promoter was too weak to express TetR in large quantity. Assembly of a stronger promoter could improve the system and lock the response to an ON/OFF response.

Besides, an experience was done to analyze the effect of the aTc inducer. Result show a visible induction of the red fluorescent protein expression by addition of aTC (60 ng/mL).



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