Team:Tokyo Tech/Project

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Project
 
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'''Code name
 
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Rising Sun'''
 
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'''''E.coli UKIYO-E'''''
 
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This project aims to make ''UKIYO-E'' by E.coli.
 
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''UKIYO-E'' is a genre of Japanese woodblock prints.
 
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{|align="center"
 
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|You can write a background of your team here.  Give us a background of your team, the members, etc.  Or tell us more about something of your choosing.
 
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|-
 
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|[[Image:tokyotech.jpg|200px|center|frame]]
 
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|-
 
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|''Tell us more about your project.  Give us background.  Use this is the abstract of your project.  Be descriptive but concise (1-2 paragraphs)''
 
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|
 
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|-
 
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|
 
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|align="center"|[[Team:Tokyo_Tech | Team Example 2]]
 
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|-
 
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|}
 
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<!--- The Mission, Experiments --->
 
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{| style="color:#1b2c8a;background-color:#0c6;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"
{| style="color:#1b2c8a;background-color:#0c6;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"
!align="center"|[[Team:Tokyo_Tech|Home]]
!align="center"|[[Team:Tokyo_Tech|Home]]
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!align="center"|[[Team:Tokyo_Tech/Team|The Team]]
 
!align="center"|[[Team:Tokyo_Tech/Project|The Project]]
!align="center"|[[Team:Tokyo_Tech/Project|The Project]]
!align="center"|[[Team:Tokyo_Tech/Parts|Parts Submitted to the Registry]]
!align="center"|[[Team:Tokyo_Tech/Parts|Parts Submitted to the Registry]]
!align="center"|[[Team:Tokyo_Tech/Modeling|Modeling]]
!align="center"|[[Team:Tokyo_Tech/Modeling|Modeling]]
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!align="center"|[[Team:Tokyo_Tech/Japanese|Japanese]]
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!align="center"|[[Team:Tokyo_Tech/Team|The Team]]
!align="center"|[[Team:Tokyo_Tech/Notebook|Notebook]]
!align="center"|[[Team:Tokyo_Tech/Notebook|Notebook]]
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[[Team:Tokyo_Tech/Mechanism|Mechanism]]
|}
|}
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(''Or you can choose different headings.  But you must have a team page, a project page, and a notebook page.'')
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          <td class="a" width="2%">　</td>
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          <td class="a">
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<p class=MsoNormal><b><font size=4>We want to make a <i style='mso-bidi-font-style:normal'>Coli Touch!!</font></b></p>
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<td class="a" width="3%">　</td>
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<!*********************************************************************************************>
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          <td class="a" width="2%">　</td>
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          <td class="a">
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<p class=MsoNormal>We try to construct a bacterial ‘touch panel’ which is colored by the pressure.</p></td>
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<td class="a" width="3%">　</td>
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          <td class="a">
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<p class=MsoNormal>We name it <i>Coli touch.</i></p></td>
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          <td class="a">
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<p class=MsoNormal>Until iGEM2007, input style was only heat, small molecule and light. These input
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style have problems. For example, Heat input occur thermal gradient, small
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molecule input occur concentration gradient, light input depend on location of a
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container. So, we thought input style except these. That is “press”. Press input
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has an advantage that input signal travel uniformly in a container.</span></p></td>
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<td class="a" width="3%">　</td>
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  </div>
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== <font size=5>'''Mechanism'''</font> ==
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<p class=MsoNormal>We construct genetic circuits that can respond to pressure signal. Lac promoter was known as a pressure inducible promoter. Lac promoter is repressed by LacI protein. The repression is released by adding IPTG (Isopropyl β-D-1-thiogalactopyranoside). Besides IPTG induction, pressure induction is possible. Under high pressure, the affinity of lac protein for lac operon decreases by transition of Lac protein from tetramer to dimer. Through this mechanism, Lac promoter is induced by pressure. </span></p></td>
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<td class="a" width="3%">　</td>
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<IMG src="https://static.igem.org/mediawiki/2008/b/bd/Test.jpg" alt="[サンプルの絵]" width="300">
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style='font-size:14.0pt'>Our project<o:p></o:p></span></u></b></p>
 
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  </div>
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lang=EN-US>We want to make a <i style='mso-bidi-font-style:normal'>Coli Touch!!</i></span></p>
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<p class=MsoNormal><span style='font-family:"ＭＳ 明朝","serif";mso-ascii-font-family:
 
-
Century;mso-hansi-font-family:Century'>→</span><span lang=EN-US>We try to
 
-
construct a bacterial ‘touch panel’ which is colored by the pressure. </span></p>
 
-
<p class=MsoNormal style='text-indent:10.5pt;mso-char-indent-count:1.0'><span
+
== <font size=5>'''Strategy'''</font> ==
-
lang=EN-US>We name it <i>Coli touch.</i></span></p>
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
<html>
 +
<body>
 +
  <div class=Section1 style='layout-grid:18.0pt'>
 +
<table width="100%" border="0" cellspacing="0" cellpadding="0">
 +
        <tr>
 +
          <td class="a" width="2%">　</td>
 +
          <td class="a">
 +
<p class=MsoNormal>We thought two projects, basic project and application project. Basic project consist of two categories. One is construction of pressure responsive genetic (1). The other is creating basic device for a touch display (2). Application project also consist of two categories. One is development of low pressure inducible promoter (3). The other is construction of pressure responsive toggle switch (4). In the future, we want to create the touch display made of E. coli.  </p></td>
 +
          <td class="a" width="3%">　</td>
 +
        </tr>
 +
      </table>
-
<! Why pressure?>
+
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<IMG src="https://static.igem.org/mediawiki/2008/c/cf/Test2.jpg" alt="[サンプルの絵]" width="700">
 +
<table width="100%" border="0" cellspacing="0" cellpadding="0">
 +
        <tr>
 +
          <td class="a" width="2%">　</td>
 +
          <td class="a">
 +
<p class=MsoNormal>We used these equipments (show figure) for pressure experiments. </p></td>
 +
          <td class="a" width="3%">　</td>
 +
        </tr>
 +
      </table>
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<IMG src="https://static.igem.org/mediawiki/2008/a/a3/Tech_kaatu.jpg" align="middle" width="500">
-
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><u><span lang=EN-US
 
-
style='font-size:14.0pt'>Why pressure?<o:p></o:p></span></u></b></p>
 
-
<p class=MsoNormal style='text-indent:5.25pt;mso-char-indent-count:.5'><span
+
  </div>
-
lang=EN-US>Example of input style to genetic circuit</span></p>
+
</body>
 +
</html>
-
<p class=MsoNormal style='text-indent:10.5pt;mso-char-indent-count:1.0'><span
+
=== <font size=5>'''Construction'''</font> ===
-
lang=EN-US>- Heat: thermal gradient </span></p>
+
-
<p class=MsoNormal style='text-indent:10.5pt;mso-char-indent-count:1.0'><span
+
<html>
-
lang=EN-US>- Small molecule: concentration gradient</span></p>
+
<body>
 +
  <div class=Section1 style='layout-grid:18.0pt'>
-
<p class=MsoNormal style='text-indent:10.5pt;mso-char-indent-count:1.0'><span
+
<table width="100%" border="0" cellspacing="0" cellpadding="0">
-
lang=EN-US>- Light: strength depend on location of container</span></p>
+
        <tr>
 +
          <td class="a" width="2%">　</td>
 +
          <td class="a">
 +
<p class=MsoNormal>We construct these plasmids (show figure). </p></td>
 +
          <td class="a" width="3%">　</td>
 +
        </tr>
 +
      </table>
-
<p class=MsoNormal><span lang=EN-US><span
+
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<IMG src="https://static.igem.org/mediawiki/2008/6/63/Tech_constraction.jpg" align="middle">
-
style='mso-spacerun:yes'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+
-
</span></span></p>
+
-
<p class=MsoNormal><span lang=EN-US><span
+
<table width="100%" border="0" cellspacing="0" cellpadding="0">
-
style='mso-spacerun:yes'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+
        <tr>
-
<IMG src="https://static.igem.org/mediawiki/2008/b/be/Tech_%E7%9F%A2%E5%8D%B0.jpg" align="middle">&nbsp;&nbsp;</span>Difficulty in uniform introduction</span></p>
+
          <td class="a" width="2%">　</td>
 +
          <td class="a">
 +
<p class=MsoNormal>Each part is constructed on low-copy plasmid “pSB6”. </p></td>
 +
          <td class="a" width="3%">　</td>
 +
        </tr>
 +
      </table>
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
  </div>
 +
</body>
 +
</html>
-
<p class=MsoNormal><span lang=EN-US><span style='mso-spacerun:yes'>&nbsp;
+
=== <font size=5>'''Basic device'''</font> ===
-
</span><span style='color:red'>Press!</span><span
+
-
style='mso-spacerun:yes'>&nbsp; </span>Confirmatory experiment </span></p>
+
-
<p class=MsoNormal><span lang=EN-US>Prospect of technological application!! </span></p>
+
<html>
 +
<body>
 +
  <div class=Section1 style='layout-grid:18.0pt'>
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
<table width="100%" border="0" cellspacing="0" cellpadding="0">
-
<!---------------------------------------------------------------------------------------------------------->
+
        <tr>  
-
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><u><span lang=EN-US>Pressure
+
          <td class="a" width="2%">　</td>
-
inducible promoter - P<sub>lac </sub><o:p></o:p></span></u></b></p>
+
          <td class="a">
 +
<p class=MsoNormal>We created pressurizing device for first step of creating touch display. This device has two holes (show figure). Each hole contains tubes and water. Inside tubes E. coli is cultivated. Pressure can travel to inside tubes. One hole (B) is covered with a block made of an acrylic glass. (show figure) Therefore the hole is not pressurized by water. The other (A) is covered with a plastic tape (show figure). Therefore the hole is pressurized.                        </p></td>
 +
          <td class="a" width="3%">　</td>
 +
        </tr>
 +
      </table>
-
<p class=MsoNormal style='text-indent:5.25pt;mso-char-indent-count:.5'><span
+
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<IMG src="https://static.igem.org/mediawiki/2008/9/9d/Tech_youki.jpg" align="middle" width="500" >
-
lang=EN-US>The affinity of LacI for the lac operator is known to decrease due
+
-
to a tetramer to dimer transition of LacI </span></p>
+
-
<IMG src="https://static.igem.org/mediawiki/2008/b/bd/Test.jpg" alt="[サンプルの絵]" width="300">
+
-
<!---------------------------------------------------------------------------------------------------------->
+
  </div>
-
<! strategy>
+
</body>
 +
</html>
 +
=== <font size=5>'''Development of low pressure inducible promoter'''</font> ===
 +
<html>
 +
<body>
 +
  <div class=Section1 style='layout-grid:18.0pt'>
-
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><u><span lang=EN-US
+
<table width="100%" border="0" cellspacing="0" cellpadding="0">
-
style='font-size:14.0pt'>Strategy<o:p></o:p></span></u></b></p>
+
        <tr>
 +
          <td class="a" width="2%">　</td>
 +
          <td class="a">
 +
<p class=MsoNormal>In the stage of basic project, very high pressure (30MPa) is required for pressure induction of lac promoter. But lower pressure response is needed for creating touch display. Therefore we develop lower pressure inducible promoter. We performed random mutagenesis to lac promoter and tried to screen promoters with FACS.</p></td>
 +
          <td class="a" width="3%">　</td>
 +
        </tr>
 +
      </table>
-
<IMG src="https://static.igem.org/mediawiki/2008/c/cf/Test2.jpg" alt="[サンプルの絵]" width="700">
+
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<IMG src="https://static.igem.org/mediawiki/2008/3/3e/Tech_dev1.jpg" align="middle" width="500" >
-
<!---------------------------------------------------------------------------------------------------------->
+
  </div>
-
<! Equipment. Instrument for pressure experiment>
+
</body>
 +
</html>
 +
=== <font size=5>'''Genetic toggle switch to implement rewritable function'''</font> ===
 +
<html>
 +
<body>
 +
  <div class=Section1 style='layout-grid:18.0pt'>
-
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><u><span lang=EN-US
+
<table width="100%" border="0" cellspacing="0" cellpadding="0">
-
style='font-size:14.0pt'>Equipment. Instrument for pressure experiment<o:p></o:p></span></u></b></p>
+
        <tr>
 +
          <td class="a" width="2%">　</td>
 +
          <td class="a">
 +
<p class=MsoNormal>A touch display are required a function of rewritable. Rewritable function consists of writable and erasable one. We adopted a genetic toggle switch that responds to pressure (show figure)</p></td>
 +
          <td class="a" width="3%">　</td>
 +
        </tr>
 +
      </table>
-
<IMG src="https://static.igem.org/mediawiki/2008/a/a3/Tech_kaatu.jpg" align="middle" width="500">
+
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<IMG src="https://static.igem.org/mediawiki/2008/5/56/Tech_4toggle1.jpg" align="middle" width="500" >
-
 
+
-
<small><p class=MsoNormal style='text-indent:5.25pt;mso-char-indent-count:.5'><span
+
-
lang=EN-US>Fig 4: Fig4-1 is a pressure vessel. Fig4-4 is a pressurizing system.
+
-
Fig4-2 show a inside of the vessel. The inside is filled with water and the
+
-
withstand pressure container is put into the inside. Fig4-3 show a water bath
+
-
placed some vessel.</span></p></small>
+
-
<!---------------------------------------------------------------------------------------------------------->
+
-
<! 1.Construction>
+
-
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><u><span lang=EN-US>Strategy
+
-
1.Construction<o:p></o:p></span></u></b></p>
+
-
 
+
-
<IMG src="https://static.igem.org/mediawiki/2008/6/63/Tech_constraction.jpg" align="middle">
+
-
 
+
-
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><span lang=EN-US>Confirmatory
+
-
experiment<o:p></o:p></span></b></p>
+
-
 
+
-
<IMG src="https://static.igem.org/mediawiki/2008/6/6b/Tech_plac_g1.jpg" align="middle">
+
-
<small><p class=MsoNormal style='text-indent:5.25pt;mso-char-indent-count:.5'><span
+
-
lang=EN-US>Fig1:Pressure-response of Plac-GFP under several pressures measured by flow cytometer</span></p></small>
+
-
 
+
-
<!---------------------------------------------------------------------------------------------------------->
+
-
<IMG src="https://static.igem.org/mediawiki/2008/e/e5/Tech_pic_30.jpg" >&nbsp;<IMG src="https://static.igem.org/mediawiki/2008/a/a2/Tech_pic_rp.jpg" height="241">&nbsp;<IMG src="https://static.igem.org/mediawiki/2008/f/f6/Tech_pic_nega.jpg" height="241">
+
-
 
+
-
<small><p class=MsoNormal style='text-indent:5.25pt;mso-char-indent-count:.5'><span
+
-
lang=EN-US>Fig2-1:Plac-GFP under 30MPa&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Fig2-2:Plac-GFP under 0.1MPa&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Fig2-3:⊿-GFP under 30MPa</span></p></small>
+
-
 
+
-
<!---------------------------------------------------------------------------------------------------------->
+
-
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><u><span lang=EN-US
+
-
style='font-size:14.0pt'>Stratagy 2. Basic device<o:p></o:p></span></u></b></p>
+
-
 
+
-
<!---------------------------------------------------------------------------------------------------------->
+
-
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><u><span lang=EN-US
+
-
style='font-size:14.0pt'>Stratagy 3. Development of low pressure inducible
+
-
promoter <o:p></o:p></span></u></b></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US>We performed random mutagenesis to lac
+
-
promoter and tried to screen promoters with FACS.</span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US>Strategy for Isolation of promoters </span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><span lang=EN-US>Future
+
-
work<o:p></o:p></span></b></p>
+
-
 
+
-
<p class=MsoNormal style='margin-left:36.0pt;text-indent:-18.0pt;mso-list:l0 level1 lfo2;
+
-
tab-stops:list 36.0pt'><![if !supportLists]><span lang=EN-US style='font-family:
+
-
"Arial","sans-serif";mso-fareast-font-family:Arial'><span style='mso-list:Ignore'>&#8226;<span
+
-
style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><![endif]><span
+
-
lang=EN-US>try Step 2 and 3</span></p>
+
-
 
+
-
<p class=MsoNormal style='margin-left:36.0pt;text-indent:-18.0pt;mso-list:l0 level1 lfo2;
+
-
tab-stops:list 36.0pt'><![if !supportLists]><span lang=EN-US style='font-family:
+
-
"Arial","sans-serif";mso-fareast-font-family:Arial'><span style='mso-list:Ignore'>&#8226;<span
+
-
style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><![endif]><span
+
-
lang=EN-US>characterize isolated promoters </span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><b style='mso-bidi-font-weight:normal'><u><span lang=EN-US>4.Genetic
+
-
toggle switch to implement rewritable function <o:p></o:p></span></u></b></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span style='font-family:"ＭＳ 明朝","serif";mso-ascii-font-family:
+
-
Century;mso-hansi-font-family:Century'>・</span><span lang=EN-US> genetic toggle
+
-
switch</span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><span
+
-
style='mso-spacerun:yes'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>- Writable
+
-
function</span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US>P<sub>lac</sub> is induced under 30 MPa
+
-
pressure</span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US>(P<sub>L </sub>is not induced by 30 MPa
+
-
pressure)</span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US>Express GFP, CI, and CI represses P<sub>L </sub></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US>→ bright !! </span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US>- Erasable function</span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US>P<sub>L</sub> is induced by heat</span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US>Express LacI, P<sub>lac </sub>represses GFP
+
-
</span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US>→ go out little by little…</span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US>Future work</span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><span
+
-
style='mso-spacerun:yes'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>- Identify
+
-
pressure model parameter from experimental data</span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><span
+
-
style='mso-spacerun:yes'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>- Simulate
+
-
pressure induction by pressure model and estimate </span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><span
+
-
style='mso-spacerun:yes'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+
-
</span>promoter-RBS strength to implement rewritable function</span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
<p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
+
-
 
+
-
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== Project Details==
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== <font size=5>'''Our Team'''</font> ==
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=== Part 2 ===
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=== The Experiments ===
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== Results ==
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== <font size=5>'''NoteBook'''</font> ==

Latest revision as of 03:40, 22 October 2008

Home The Project Parts Submitted to the Registry Modeling Japanese The Team Notebook

Mechanism


Contents

Our project

　

We want to make a Coli Touch!!

　
　

We try to construct a bacterial ‘touch panel’ which is colored by the pressure.

 　
　

We name it Coli touch.

 　
　

Until iGEM2007, input style was only heat, small molecule and light. These input style have problems. For example, Heat input occur thermal gradient, small molecule input occur concentration gradient, light input depend on location of a container. So, we thought input style except these. That is “press”. Press input has an advantage that input signal travel uniformly in a container.

 　

Mechanism

　

We construct genetic circuits that can respond to pressure signal. Lac promoter was known as a pressure inducible promoter. Lac promoter is repressed by LacI protein. The repression is released by adding IPTG (Isopropyl β-D-1-thiogalactopyranoside). Besides IPTG induction, pressure induction is possible. Under high pressure, the affinity of lac protein for lac operon decreases by transition of Lac protein from tetramer to dimer. Through this mechanism, Lac promoter is induced by pressure.

　
          [サンプルの絵]


Strategy

　

We thought two projects, basic project and application project. Basic project consist of two categories. One is construction of pressure responsive genetic (1). The other is creating basic device for a touch display (2). Application project also consist of two categories. One is development of low pressure inducible promoter (3). The other is construction of pressure responsive toggle switch (4). In the future, we want to create the touch display made of E. coli.

　
          [サンプルの絵]
　

We used these equipments (show figure) for pressure experiments.

　
          

Construction

　

We construct these plasmids (show figure).

　
          
　

Each part is constructed on low-copy plasmid “pSB6”.

　

Basic device

　

We created pressurizing device for first step of creating touch display. This device has two holes (show figure). Each hole contains tubes and water. Inside tubes E. coli is cultivated. Pressure can travel to inside tubes. One hole (B) is covered with a block made of an acrylic glass. (show figure) Therefore the hole is not pressurized by water. The other (A) is covered with a plastic tape (show figure). Therefore the hole is pressurized.

　
          

Development of low pressure inducible promoter

　

In the stage of basic project, very high pressure (30MPa) is required for pressure induction of lac promoter. But lower pressure response is needed for creating touch display. Therefore we develop lower pressure inducible promoter. We performed random mutagenesis to lac promoter and tried to screen promoters with FACS.

 　
          

Genetic toggle switch to implement rewritable function

　

A touch display are required a function of rewritable. Rewritable function consists of writable and erasable one. We adopted a genetic toggle switch that responds to pressure (show figure)

 　
          

Our Team

NoteBook

Retrieved from "http://2008.igem.org/Team:Tokyo_Tech/Project"