Team:Chiba/Modeling

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{| style="color:#dd0000;background-color:#eeeeee;" cellpadding="3" cellspacing="1" border="1" bordercolor="#cc0303" width="62%" align="center"
 
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!align="center"|[[Team:Chiba|Home]]
 
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!align="center"|[[Team:Chiba/Team|Team]]
 
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!align="center"|[[Team:Chiba/Project|Project]]
 
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!align="center"|[[Team:Chiba/Parts|Parts]]
 
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!align="center"|[[Team:Chiba/Modeling|Modeling]]
 
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!align="center"|[[Team:Chiba/Notebook|Notebook]]
 
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!align="center"|[[Team:Chiba/Safety|Safety]]
 
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Our second design of genetic double-click system consists of a pulse-generator and two inverters which can be seemed as a slow pulse.<br>
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In this system, we also use AHL input and GFP output.<br>
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This time, we use AHL as an activate signal, so when there is AHL added, Lux promoter will be activated.<br>
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The transcription factors of output are T7 RNA Polymerase and cI repressor. <br>
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At initial state, LuxR and cI protein are constitutively generated.<br>
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So cI binds to the operator site of PT7/cI.<br>
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When 1st input is injected, the LuxR-AHL dimmer binds to the Lux-box of the lux promoters so that T7 RNA Polymerase,<br>
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cI434 and tetR protein are generated at the same time.<br>
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In a while, cI434 protein will be repress the transcription of T7 RNA Polymerase so that the expression of T7 RNA Polymerase can be shown as a pulse.<br>
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At the same time. tetR begin to repress cI expression, but the time of cI gets lost will be take longer then T7 RNA Polymerase pulse so that there will be no<br>
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GFP output.<br><br>
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C2 gradually accumulates, it binds the operator sites of the Plux/c2 promoter and stops transcription of T7 RNAP.<br>
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So, T7 RNAP is generated as a pulse.<br>
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At the same time, tetR binds the tetR operator sites of the tetR promoter (Ptet) as a dimer.<br>
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Transcription of CI is stopped by tetR, CI decomposes and the PT7/CI promoter is unbound.<br>
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This derepression occurs after the pulse of T7 RNAP has passed.<br>
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In other words, the operator sites of PT7/CI are bound by CI when there is pulse of T7 RNAP.<br>
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So, it cannot transcribe GFP. TetR creates a time delay from input to derepression.<br>
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Thanks to this time delay and one-time pulse, bacteria can never shine by an only first input. <br><br>
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Before injecting 2nd input, we must create none-input-environment.<br>
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So we choose to wash the 1st input. By washing , tetR protein and cI434 protein will degrade.<br>
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cI434 protein should disappear so that when there is 2nd input, the T7 RNA Polymerase will be shown as a pulse same with the 1st time.<br>
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cI will begin to generate if tetR protein gets lost.<br>
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We recognize it as time-limit, when there is enough cI generated (this means cI repress is stronger than T7 RNA Polymerase),<br>
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there will be no GFP output.<br>
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On the contrary, when there is less cI protein or no cI protein at the moment, T7 RNA Polymerase pulse can activate GFP output. <br><br>
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By the second injecting AHL before the inhibition by CI, T7 RNAP binds the PT7/CI promoter and transcribes the downstream GFP.<br><br>
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<li><a href="https://2010.igem.org/Team:Chiba"><span>Home</span></a></li>
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<li><a href="https://2010.igem.org/Team:Chiba/Team"><span>Team</span></a>
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<ul style="z-index:1"> <br>
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<li><a class="tablinks" href="https://2010.igem.org/Team:Chiba/Team">Team</a></li> <li><a class="tablinks" href="https://2010.igem.org/Team:Chiba/Member">Member</a></li>
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<li><a class="tablinks" href="https://2010.igem.org/Team:Chiba/School">School</a></li>
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</ul>
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</li>
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<li><a href="https://2010.igem.org/Team:Chiba/Project"><span>Project</span></a>
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<ul style="z-index:1"><br>
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                        <li><a class="tablinks" href="https://2010.igem.org/Team:Chiba/Project">Overall Project</a></li>
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<li><a class="tablinks" href="https://2010.igem.org/Team:Chiba/Circuit_1">Circuit 1</a></li>
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<li><a class="tablinks" href="https://2010.igem.org/Team:Chiba/Circuit_2">Circuit 2</a></li>
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</ul>
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</li>
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<li><a href="https://2010.igem.org/Team:Chiba/Parts"><span>Parts</span></a></li>
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<li><a href="https://2010.igem.org/Team:Chiba/Modeling"><span>Modeling</span></a></li>
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<li><a href="https://2010.igem.org/Team:Chiba/Result"><span>Result</span></a>
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<ul style="z-index:1"><br>
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                        <li><a class="tablinks" href="https://2010.igem.org/Team:Chiba/Plasmid1">Plasmid 1</a></li>
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<li><a class="tablinks" href="https://2010.igem.org/Team:Chiba/Plasmid2">Plasmid 2</a></li>
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<li><a class="tablinks" href="https://2010.igem.org/Team:Chiba/lux_promoter">lux Promoter</a></li><br>
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</ul>
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</li>
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<li><a href="https://2010.igem.org/Team:Chiba/Notebook"><span>Notebook</span></a>
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<ul style="z-index:1"><br>
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                        <li><a class="tablinks" href="https://2010.igem.org/Team:Chiba/Notebook">Notebook 1</a></li>
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<li><a class="tablinks" href="https://2010.igem.org/Team:Chiba/Notebook_2">Notebook 2</a></li>
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<li><a class="tablinks" href="https://2010.igem.org/Team:Chiba/Notebook_3">Notebook 3</a></li><br>
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</ul>
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</li>
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<li><a href="https://2010.igem.org/Team:Chiba/Support"><span>Support</span></a></li>
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<li><a href="https://2010.igem.org/Team:Chiba/Safety"><span>Safety</span></a></li>
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Latest revision as of 04:44, 25 October 2010




 




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