Team:Chiba/Project

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<font color="black" size="5.5">Bacterial Double Click</font>
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----
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<img src="https://static.igem.org/mediawiki/2010/f/f9/Chiba_menu_2.jpg">
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If there are two-time input for limited time, bacteria shine.
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== '''Overall project''' ==
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|We’re inspired by double-click of computer’s mouse.<br>
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It doesn’t react by first input but react by second input. So we designed bacterial DNA sequence which works like it. Furthermore, we built time-limit in genetic sequence between the first and the second input like a computer mouse. Bacteria which have the DNA sequence can distinguish between the first input and the second one. Once in a while, bacteria might receive input which wasn’t intended. But it was the first input so bacteria don’t react. Even if there was input not intended, bacteria don’t react unless the second input is entered. Moreover, when the limit-time which controlled by the genetic system is passed, the system is going to return to the initial state. So we expect this DNA sequence will work as a safety device.
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|align="center"| [[Image:click.jpg|200px|left|frame]]
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== Project Details==
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<li id="current"><a href="https://2010.igem.org/Team:Chiba/Safety"><span>Safety</span></a></li>
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===Requirement of Double Click ===
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1. If there is a click only one-time, nothing happens.<br>
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2. Time of clicking button doesn't matter. It just depends on the number of click. One or two.<br>
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3. After a lot of time pass from the 1st click, it will return to the initial state.<br>
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4. Click two-time in specific limited time(We call this double click.), something occur.
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=== Explanation of DNA sequence ===
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[[Image:Chiba_model_1.jpg‎]]
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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First, we use AHL for input and regard injection of AHL as clicking. <br>
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LuxR is generated constitutive.<br>
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Injecting AHL to Double Click Bacteria brings dimer of AHL and luxR.&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<br>
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We take lux promoter repressed by the dimer.<br>
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As the dimer binds Plux, transcription of proteins under the<br>
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Plux is repressed.<br><br>
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So in our project, <br>
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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<b><font color="red">CLICK controls transcription of overall DNA sequence.</font></b>
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|align="center"| [[Image:Chiba_LuxR.jpg‎|400px|left|frame]]
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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Second, there is a cI operator above gfp DNA sequence.<br>
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If cI
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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cI works as repressor.<br>
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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You can see cI DNA sequence is under the lux promoter.<br>
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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As AHL come into Double Click Bacteria, generation of cI is stopped.&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<br>
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Therefore, repressor of cI operator disappears.<br>
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(Because the cause of repress, cI, which is kind of protein will<br>
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be decomposed.)<br>
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If there isn't cI anymore, bacteria will shine with gfp.<br><br>
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We can say<br>
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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<b><font color="red">cI recognizes the existence of input in this DNA sequence.</font></b>
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|align="center"| [[Image:Chiba_cI.jpg|400px|left|frame]]
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Third, there is an AND Gate with T7ptag and supD.<br>
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As you know,.<br>
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T7ptag becomes T7 polymerase in case of existing supD.<br>
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T7 polymerase is translated to T7 protein and T7 works as activator<br>
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of T7 promoter.<br>
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There is a T7 promoter above gfp DNA sequence.<br>
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If T7 have been generated once, the T7 promoter starts translating.<br>
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And then we can see bacteria shine with gfp.<br>
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On initial state(there isn't any click), bacteria don't have supD.<br>
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So there isn't any T7 polymerase and gfp cannot be genernated,<br>
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in spite of being T7ptag in bacteria.<br>
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T7ptag which is a kind of mRNA is decomposed so fast.<br>
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However supD under lacI promoter is generated late.<br>
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Because, transcription of supD starts working after most of lacI are<br>
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decomposed.<br>
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After 1st click, supD exist.(supD is a kind of tRNA which is stable.)<br>
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There isn't input anymore,<br>
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promoter above T7ptag DNA sequence starts activating.<br>
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As a result of reacting T7ptag and supD,<br>
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the AND Gate is going to be available.<br>
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In this case, the AND Gate remembers that  <b><font color="red">there was a click.</font></b><br>
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Moreover,
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|align="center"| [[Image:Chiba_third.jpg‎|400px|left|frame]]
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                <ul>
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                                <!-- CSS Tabs -->
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<li><a href="https://2010.igem.org/Team:Chiba/Project"><span>Double Click System</span></a></li>
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<li><a href="https://2010.igem.org/Team:Chiba/System_1"><span>Version 1</span></a></li>
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<li><a href="https://2010.igem.org/Team:Chiba/System_2"><span>Version 2</span></a></li>                      </ul>
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</html>
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<!--- 2nd tab End--->
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<!--- Background Contents End --->
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<!-- -------------------------------------------------------------------------------
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-------------------------------------------------------------------------------
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ここから下を編集
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-------------------------------------------------------------------------------
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----------------------------------------------------------------------------------- -->
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<br><br><br>
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===<font size="5">Double Click Bacteria</font>===
 +
-----
 +
We’re inspired by double-click of computer’s mouse. It doesn’t react to the first click but does react when it is accompanied by the second one. This is one of the most accepted, familiarized, and proven mechanism to diminish the erroneous operation. This fail-safe technology should find various uses also in biotechnology.
 +
Here is what we envision: once in a while, bacteria might receive input which wasn’t intended. But they are careful enough (they know Homo sapiens always make mistake!) so that they ignore the input for the first time.  However, when they receive the two inputs in succession, they finally judge the input(s) are real, and start taking the action as programmed.  Also, they are smart enough to distinguish double-clicking from the two separate (or erroneous) inputs.
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<br><br><br>
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First, the circuit must distinguish the second input (stimulus) from the first one, even though they are chemically physically the same . To realize this, we need the counting device. Second, the circuit must distinguishes W-click from two separate clicks; the circuit reacts only when the first and the second input are given in short period of time. Thus, the genetic counter should be coupled with the genetic timers. After a certain time after the first input (timeout), the circuit gets back to the initial state so that the  next stimulous is recognized as the first input.
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<html>
<center>
<center>
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[[Image:Chiba_model_4.jpg‎]]
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Actually, there is a hybrid promoter(cI/T7) above gfp DNA sequence.<br>
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T7 works as activator and cI works as repressor to the promoter.<br>
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=== Part 3 ===
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===<font size="5">The Bottom Line is...</font>===
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#If there is only an input, nothing happens.
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#Duration of the input is not the matter. The circuit cares only the number of input.
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#However, a certain time after the 1st input, it returns to the initial state.
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#Giving two inputs in the limited time the circuit get activated (gives output).
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== Results ==
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===<font size="5">Systems</font>===
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<a href="https://2010.igem.org/Team:Chiba/System_1"><img src="https://static.igem.org/mediawiki/2010/d/db/Chiba_plan2_1.jpg" border="0" width="300px" /><br>version 1</a>
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<a href="https://2010.igem.org/Team:Chiba/System_2"><img src="https://static.igem.org/mediawiki/2010/1/1a/Chiba_Sys2.jpg" border="0" width="300px" /><br>version 2</a>
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Latest revision as of 03:10, 28 October 2010




 

 




Double Click Bacteria


We’re inspired by double-click of computer’s mouse. It doesn’t react to the first click but does react when it is accompanied by the second one. This is one of the most accepted, familiarized, and proven mechanism to diminish the erroneous operation. This fail-safe technology should find various uses also in biotechnology.

Here is what we envision: once in a while, bacteria might receive input which wasn’t intended. But they are careful enough (they know Homo sapiens always make mistake!) so that they ignore the input for the first time. However, when they receive the two inputs in succession, they finally judge the input(s) are real, and start taking the action as programmed. Also, they are smart enough to distinguish double-clicking from the two separate (or erroneous) inputs.

First, the circuit must distinguish the second input (stimulus) from the first one, even though they are chemically physically the same . To realize this, we need the counting device. Second, the circuit must distinguishes W-click from two separate clicks; the circuit reacts only when the first and the second input are given in short period of time. Thus, the genetic counter should be coupled with the genetic timers. After a certain time after the first input (timeout), the circuit gets back to the initial state so that the next stimulous is recognized as the first input.




The Bottom Line is...

  1. If there is only an input, nothing happens.
  2. Duration of the input is not the matter. The circuit cares only the number of input.
  3. However, a certain time after the 1st input, it returns to the initial state.
  4. Giving two inputs in the limited time the circuit get activated (gives output).



Systems


version 1

version 2