Team:Tokyo Tech
From 2008.igem.org
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<div class=Section1 style='layout-grid:18.0pt'> | <div class=Section1 style='layout-grid:18.0pt'> | ||
- | <p class=MsoNormal><b | + | <p class=MsoNormal><b><u><font size=4>Our project</font></u></b></p> |
- | style='font- | + | <table width="100%" border="0" cellspacing="0" cellpadding="0"> |
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+ | <td class="a" width="0%"> </td> | ||
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+ | <p class=MsoNormal><b><u><font size=4>We want to make a <i style='mso-bidi-font-style:normal'>Coli Touch!!</font></u></b></p> | ||
+ | <td class="a" width="3%"> </td> | ||
+ | </tr> | ||
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- | + | <p class=MsoNormal>We try to construct a bacterial ‘touch panel’ which is colored by the pressure.</p></td> | |
- | + | <td class="a" width="3%"> </td> | |
+ | </tr> | ||
+ | </table> | ||
+ | <table width="100%" border="0" cellspacing="0" cellpadding="0"> | ||
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<p class=MsoNormal style='text-indent:10.5pt;mso-char-indent-count:1.0'><span | <p class=MsoNormal style='text-indent:10.5pt;mso-char-indent-count:1.0'><span | ||
- | lang=EN-US>We name it <i>Coli touch.</i></span></p> | + | lang=EN-US>We name it <i>Coli touch.</i></span></p></td> |
+ | <td class="a" width="3%"> </td> | ||
+ | </tr> | ||
+ | </table> | ||
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molecule input occur concentration gradient, light input depend on location of a | 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 | container. So, we thought input style except these. That is “press”. Press input | ||
- | has an advantage that input signal travel uniformly in a container.</ | + | has an advantage that input signal travel uniformly in a container.</span></p></td> |
<td class="a" width="3%"> </td> | <td class="a" width="3%"> </td> | ||
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- | <p class=MsoNormal><b | + | <p class=MsoNormal><b><u><font size=4>Mechanism</font></u></b></p> |
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+ | <table width="100%" border="0" cellspacing="0" cellpadding="0"> | ||
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<p class=MsoNormal style='text-indent:5.25pt;mso-char-indent-count:.5'><span | <p class=MsoNormal style='text-indent:5.25pt;mso-char-indent-count:.5'><span | ||
- | lang=EN-US>The affinity of | + | lang=EN-US>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> |
- | + | <td class="a" width="3%"> </td> | |
+ | </tr> | ||
+ | </table> | ||
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<IMG src="https://static.igem.org/mediawiki/2008/b/bd/Test.jpg" alt="[サンプルの絵]" width="300"> | <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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+ | <p class=MsoNormal><b><u><font size=4>Strategy</font></u></b></p> | ||
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+ | <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> | ||
<IMG src="https://static.igem.org/mediawiki/2008/c/cf/Test2.jpg" alt="[サンプルの絵]" width="700"> | <IMG src="https://static.igem.org/mediawiki/2008/c/cf/Test2.jpg" alt="[サンプルの絵]" width="700"> | ||
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- | + | <p class=MsoNormal>We used these equipments (show figure) for pressure experiments. </p></td> | |
- | < | + | <td class="a" width="3%"> </td> |
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+ | </table> | ||
<IMG src="https://static.igem.org/mediawiki/2008/a/a3/Tech_kaatu.jpg" align="middle" width="500"> | <IMG src="https://static.igem.org/mediawiki/2008/a/a3/Tech_kaatu.jpg" align="middle" width="500"> | ||
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- | + | <p class=MsoNormal><b><u><font size=4>1.Construction</font></u></b></p> | |
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- | 1.Construction< | + | <table width="100%" border="0" cellspacing="0" cellpadding="0"> |
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+ | <p class=MsoNormal>We construct these plasmids (show figure). </p></td> | ||
+ | <td class="a" width="3%"> </td> | ||
+ | </tr> | ||
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<IMG src="https://static.igem.org/mediawiki/2008/6/63/Tech_constraction.jpg" align="middle"> | <IMG src="https://static.igem.org/mediawiki/2008/6/63/Tech_constraction.jpg" align="middle"> | ||
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- | + | <p class=MsoNormal>Each part is constructed on low-copy plasmid “pSB6”. </p></td> | |
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+ | <p class=MsoNormal><b><u><font size=4>2. Basic device</font></u></b></p> | ||
- | < | + | <table width="100%" border="0" cellspacing="0" cellpadding="0"> |
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- | + | <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> |
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<IMG src="https://static.igem.org/mediawiki/2008/9/9d/Tech_youki.jpg" align="middle" width="500" > | <IMG src="https://static.igem.org/mediawiki/2008/9/9d/Tech_youki.jpg" align="middle" width="500" > | ||
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- | <p class=MsoNormal><b | + | <p class=MsoNormal><b><u><font size=4>3. Development of low pressure inducible promoter </font></u></b></p> |
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- | + | <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> | |
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- | + | <p class=MsoNormal><b><u><font size=4>4.Genetic toggle switch to implement rewritable function </font></u></b></p> | |
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<p class=MsoNormal><span lang=EN-US><o:p> </o:p></span></p> | <p class=MsoNormal><span lang=EN-US><o:p> </o:p></span></p> |
Revision as of 23:37, 21 October 2008
Home | The Team | The Project | Parts Submitted to the Registry | Modeling | Notebook | Japanese |
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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]](https://static.igem.org/mediawiki/2008/b/bd/Test.jpg)
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]](https://static.igem.org/mediawiki/2008/c/cf/Test2.jpg)
We used these equipments (show figure) for pressure experiments. |
![](https://static.igem.org/mediawiki/2008/a/a3/Tech_kaatu.jpg)
1.Construction
We construct these plasmids (show figure). |
![](https://static.igem.org/mediawiki/2008/6/63/Tech_constraction.jpg)
Each part is constructed on low-copy plasmid “pSB6”. |
2. 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. |
![](https://static.igem.org/mediawiki/2008/9/9d/Tech_youki.jpg)
3. 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. |
![](https://static.igem.org/mediawiki/2008/3/3e/Tech_dev1.jpg)
4.Genetic toggle switch to implement rewritable function
・ genetic toggle switch
- Writable function
Plac is induced under 30 MPa pressure
(PL is not induced by 30 MPa pressure)
Express GFP, CI, and CI represses PL
→ bright !!
- Erasable function
PL is induced by heat
Express LacI, Plac represses GFP
→ go out little by little…
Future work
- Identify pressure model parameter from experimental data
- Simulate pressure induction by pressure model and estimate
promoter-RBS strength to implement rewritable function
Mathematical model