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- | <p><span style="font-style:italic;">Signalling is nothing without control...</span></p> | + | <table align='center' width='100%'> |
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| + | <img alt="" src="https://static.igem.org/mediawiki/2011/b/b1/UNIPV_Pavia_Work_in_progress_scribblings.jpg" class="thumbimage" width="93%"></a></div></div> |
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| + | <h2 class="art-postheader">The project</h2> |
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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. |
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| + | <img src="https://static.igem.org/mediawiki/2011/2/2d/UNIPV_Hometeam1.jpg" title="UNIPV iGEM team" alt="" /> |
| + | <img src="https://static.igem.org/mediawiki/2011/0/0a/UNIPV_Hometeam2.jpg" title="UNIPV iGEM team" alt="" /> |
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- | <p style="text-align:justify;">One of the pivotal objectives of synthetic biology is to build complex gene networks with a predictable behavior by combining well-characterized basic modules. </p> | + | </div> |
- | <p style="text-align:justify;">As a proof of concept of this fundamental, our project aims at designing and implementing in <em>E. coli</em> a quorum sensing-based control system, able to regulate the concentration of a signaling molecule (3OC6-HSL) via a negative feedback loop. </p> | + | </div> |
- | <p style="text-align:justify;">In order to obtain the desired output, fine-tuning of the circuit is necessary; therefore, a mathematical model of the control system will be derived and identified by data coming from <em>ad hoc</em> experiments performed on basic modules. Model simulations will be used to meet the design specifications of the biological controller, demonstrating that full characterization of basic parts is a major goal to predict the behavior of more complex circuits.</p> | + | <div id="nivoslider-125-caption-0" class="nivo-html-caption">You can add captions too…</div> |
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