Team:UNIPV-Pavia

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<div style="text-align:center"><div class="thumbinner" style="width: 800px;"><img src="https://static.igem.org/mediawiki/2011/b/b1/UNIPV_Pavia_Work_in_progress_scribblings.jpg" class="thumbimage" width="89%"></a></div></div>
<div style="text-align:center"><div class="thumbinner" style="width: 800px;"><img src="https://static.igem.org/mediawiki/2011/b/b1/UNIPV_Pavia_Work_in_progress_scribblings.jpg" class="thumbimage" width="89%"></a></div></div>
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<p align='justify'>Our work aims at implementing the engineering concept of closed-loop control in E. coli, exploiting quorum sensing. As a proof of concept, we designed a simple genetic controller that regulates the concentration of 3OC6-HSL signalling molecule around  a user-defined set-point. The controlled variable (3OC6-HSL) increases as a function of the exogenous anhydroTetracycline input, that triggers LuxI expression. The controller senses the 3OC6-HSL concentration and activates the production of AiiA, that degrades it.
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<p align='justify'>Our work aims at implementing the engineering concept of closed-loop control in <em>E. coli</em>, exploiting quorum sensing. As a proof of concept, we designed a simple genetic controller that regulates the concentration of 3OC6-HSL signalling molecule around  a user-defined set-point. The controlled variable (3OC6-HSL) increases as a function of the exogenous anhydroTetracycline input, that triggers LuxI expression. The controller senses the 3OC6-HSL concentration and activates the production of AiiA, that degrades it.
To observe the desired behaviour, a fine tuning of the system was necessary.  The transcriptional/translational strength of the regulatory elements (promoter+RBS in several combinations) and the enzyme activities were measured and exploited to identify a mathematical model able to predict the behaviour of the controlled system. These predictions made possible an in silico rational fine tuning of the circuit: the most promising modules were selected and assembled into the final circuit, avoiding a cost and time expensive combinatorial approach.
To observe the desired behaviour, a fine tuning of the system was necessary.  The transcriptional/translational strength of the regulatory elements (promoter+RBS in several combinations) and the enzyme activities were measured and exploited to identify a mathematical model able to predict the behaviour of the controlled system. These predictions made possible an in silico rational fine tuning of the circuit: the most promising modules were selected and assembled into the final circuit, avoiding a cost and time expensive combinatorial approach.
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Revision as of 11:08, 17 September 2011

UNIPV TEAM 2011

Abstract

Our work aims at implementing the engineering concept of closed-loop control in E. coli, exploiting quorum sensing. As a proof of concept, we designed a simple genetic controller that regulates the concentration of 3OC6-HSL signalling molecule around a user-defined set-point. The controlled variable (3OC6-HSL) increases as a function of the exogenous anhydroTetracycline input, that triggers LuxI expression. The controller senses the 3OC6-HSL concentration and activates the production of AiiA, that degrades it. To observe the desired behaviour, a fine tuning of the system was necessary. The transcriptional/translational strength of the regulatory elements (promoter+RBS in several combinations) and the enzyme activities were measured and exploited to identify a mathematical model able to predict the behaviour of the controlled system. These predictions made possible an in silico rational fine tuning of the circuit: the most promising modules were selected and assembled into the final circuit, avoiding a cost and time expensive combinatorial approach.

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