Difference between revisions of "Team:Fudan/Basic Part"

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        <a href="https://2017.igem.org/Team:Fudan/Demonstrate"><img class="align" id="align-1" src='https://static.igem.org/mediawiki/2017/9/93/T--Fudan--synNotch.png' /></a>
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        <a href="https://2017.igem.org/Team:Fudan/Basic_Part"><img class="align" id="align-4" src='https://static.igem.org/mediawiki/2017/2/24/T--Fudan--hccOrange.png' /></a>
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  <h class="sup-title dark-blue"></br>Basic Part – surEGFP</br></h>
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<div class="block main-text main-text">
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  <h class="highlight-title dark-blue"></br></br>For this year’s iGEM competition, our favorite basic part is surEGFP </br>(BBa_K2446051) </h>
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  <p></br>This part is a surface-expressed EGFP which can be used to create heterogeneous antigen density expressing cell model. Our team is very excited to be the first to bring the idea of antigen density heterogeneity to the iGEM competition, and propose a tri-response immunotherapy, <a href="https://2017.igem.org/Team:Fudan/Applied_Design">SwordS</a>.
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<div class="block main-text main-text">
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  <h class="highlight-title dark-blue" id="Antigen"></br></br>How surEGFP works</h>
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  <p>Surface-expressed EGFP (surEGFP) is built by fusing CD8α signal peptid and EGFP to the transmembrane region of CD86 (NM_175862) from N terminal to C terminal.
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CD8α peptide guides EGFP to be translocated via <a target="_blank" href="https://en.wikipedia.org/wiki/Sec61">Sec61 into the <a href="https://en.wikipedia.org/wiki/Endoplasmic_reticulum" target="_blank">endoplasmic reticulum (ER)</a> lumen, while transmembrane region of CD86 embeds EGFP on the plasma membrane. Below, two confocal optical slices with 0.5 micron apart are shown, and the yellow arrowheads point to ER lumen/membrane with surEGFP. EGFP bleach really fast in the cell culture media and we could obtain a strong signal after surEGFP export out of the cell and expose to the external environment. We confirmed the cell surface exposed surEGFP by immunostaining with antibody against GFP, without permeabilizing the cells.
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<img width="60%" src="https://static.igem.org/mediawiki/2017/7/74/T--Fudan--BPFigure.png">
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<div class="block main-text main-text">
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  <h class="highlight-title dark-blue" id="Antigen"></br></br>See More</h>
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<p>
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Integrated information about surEGFP posted on <a href="https://2017.igem.org/Team:Fudan/Demonstrate">Demonstrate</a> page: Antigen density heterogeneity cell model. Or you can alternatively check the <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2446051">(BBa_K2446051)</a> on the Registry of Standard Biological Parts website.
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</p>
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<p>
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We listed other basic part we constructed this year below.
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</p>
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    <td>2_ZF42-10/43-8 binding site</td>
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    <td ><a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K2446001>(BBa_K2446001)</a></td>
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  </tr>
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  <tr>
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    <td>2_ZF54-8/43-8 binding site</td>
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    <td><a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K2446002>(BBa_K2446002)</a></td>
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  </tr>
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  <tr>
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    <td>2_ZFHD1 binding site</td>
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    <td><a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K2446003>(BBa_K2446003)</a></td>
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  </tr>
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  <tr>
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    <td>2_PIP binding site</td>
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    <td><a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K2446004>(BBa_K2446004)</a></td>
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  </tr>
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    <td>4_ZF_42-10 binding site</td>
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    <td><a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K2446005>(BBa_K2446005)</a></td>
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  </tr>
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    <td>4_ZF_43-8 binding site</td>
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    <td><a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K2446006>(BBa_K2446006)</a></td>
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  </tr>
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    <td>4_ZF_54-8 binding site</td>
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    <td><a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K2446007>(BBa_K2446007)</a></td>
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  </tr>
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    <td>4_ZF_21-16 binding site</td>
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    <td><a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K2446008>(BBa_K2446008)</a></td>
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  </tr>
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    <td>4_ZF54-8/43-8 binding site</td>
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    <td><a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K2446009>(BBa_K2446009)</a></td>
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  </tr>
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  <tr>
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    <td>4_ZFHD1 binding site</td>
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    <td><a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K2446010>(BBa_K2446010)</a></td>
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  </tr>
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  <tr>
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    <td>4_PIP binding site</td>
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    <td><a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K2446011>(BBa_K2446011)</a></td>
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  </tr>
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  <tr>
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    <td>8_ZF_42-10 binding site</td>
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    <td><a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K2446012>(BBa_K2446012)</a></td>
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  </tr>
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  <tr>
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    <td>8_ZF_43-8 binding site</td>
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    <td><a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K2446013>(BBa_K2446013)</a></td>
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  </tr>
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  <tr>
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    <td>8_ZF_54-8 binding site</td>
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    <td><a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K2446014>(BBa_K2446014)</a></td>
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  </tr>
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  <tr>
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    <td>8_ZF_21-16 binding site</td>
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    <td><a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K2446015>(BBa_K2446015)</a></td>
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  </tr>
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  <tr>
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    <td>8_ZF_54-8/43-8 binding site</td>
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    <td><a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K2446016>(BBa_K2446016)</a></td>
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  </tr>
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  <tr>
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    <td>8_PIP binding site</td>
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    <td><a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K2446017>(BBa_K2446017)</a></td>
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  </tr>
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  <tr>
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    <td>ZF_GAl4_KRAB</td>
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    <td><a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K2446037>(BBa_K2446037)</a></td>
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  </tr>
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  <tr>
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    <td>ZF_PIP_KRAB</td>
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    <td><a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K2446038>(BBa_K2446038)</a></td>
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  </tr>
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  <tr>
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    <td>ZF_21-16KRAB</td>
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    <td><a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K2446039>(BBa_K2446039)</a></td>
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  </tr>
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  <tr>
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    <td>ZF_42-10_KRAB</td>
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    <td><a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K2446040>(BBa_K2446040)</a></td>
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  </tr>
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  <tr>
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    <td>ZF_43-8_KRAB</td>
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    <td><a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K2446041>(BBa_K2446041)</a></td>
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  </tr>
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  <tr>
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    <td>ZF_54-8_KRAB</td>
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    <td><a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K2446042>(BBa_K2446042)</a></td>
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  </tr>
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  <tr>
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    <td>ZFHD1_KRAB</td>
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    <td><a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K2446043>(BBa_K2446043)</a></td>
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  </tr>
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<p></br></br></p>
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{{Fudan/PageFooter}}
 
{{Fudan/PageFooter}}

Latest revision as of 01:22, 2 November 2017

Basic Part – surEGFP


For this year’s iGEM competition, our favorite basic part is surEGFP
(BBa_K2446051)


This part is a surface-expressed EGFP which can be used to create heterogeneous antigen density expressing cell model. Our team is very excited to be the first to bring the idea of antigen density heterogeneity to the iGEM competition, and propose a tri-response immunotherapy, SwordS.



How surEGFP works

Surface-expressed EGFP (surEGFP) is built by fusing CD8α signal peptid and EGFP to the transmembrane region of CD86 (NM_175862) from N terminal to C terminal. CD8α peptide guides EGFP to be translocated via Sec61 into the endoplasmic reticulum (ER) lumen, while transmembrane region of CD86 embeds EGFP on the plasma membrane. Below, two confocal optical slices with 0.5 micron apart are shown, and the yellow arrowheads point to ER lumen/membrane with surEGFP. EGFP bleach really fast in the cell culture media and we could obtain a strong signal after surEGFP export out of the cell and expose to the external environment. We confirmed the cell surface exposed surEGFP by immunostaining with antibody against GFP, without permeabilizing the cells.



See More

Integrated information about surEGFP posted on Demonstrate page: Antigen density heterogeneity cell model. Or you can alternatively check the (BBa_K2446051) on the Registry of Standard Biological Parts website.

We listed other basic part we constructed this year below.

2_ZF42-10/43-8 binding site (BBa_K2446001)
2_ZF54-8/43-8 binding site (BBa_K2446002)
2_ZFHD1 binding site (BBa_K2446003)
2_PIP binding site (BBa_K2446004)
4_ZF_42-10 binding site (BBa_K2446005)
4_ZF_43-8 binding site (BBa_K2446006)
4_ZF_54-8 binding site (BBa_K2446007)
4_ZF_21-16 binding site (BBa_K2446008)
4_ZF54-8/43-8 binding site (BBa_K2446009)
4_ZFHD1 binding site (BBa_K2446010)
4_PIP binding site (BBa_K2446011)
8_ZF_42-10 binding site (BBa_K2446012)
8_ZF_43-8 binding site (BBa_K2446013)
8_ZF_54-8 binding site (BBa_K2446014)
8_ZF_21-16 binding site (BBa_K2446015)
8_ZF_54-8/43-8 binding site (BBa_K2446016)
8_PIP binding site (BBa_K2446017)
ZF_GAl4_KRAB (BBa_K2446037)
ZF_PIP_KRAB (BBa_K2446038)
ZF_21-16KRAB (BBa_K2446039)
ZF_42-10_KRAB (BBa_K2446040)
ZF_43-8_KRAB (BBa_K2446041)
ZF_54-8_KRAB (BBa_K2446042)
ZFHD1_KRAB (BBa_K2446043)