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− | {{IGEM_TopBar}}
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− | {{Shanghai_Metropolis}}
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| <html> | | <html> |
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| + | <link rel="stylesheet" type="text/css" |
| + | href="https://2021.igem.org/wiki/index.php?title=Template:Shanghai_Metropolis/StyleCss&action=raw&ctype=text/css"/> |
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− | <div class="column full_size judges-will-not-evaluate"> | + | <div class="ev71-content"> |
− | <h3>★ ALERT! </h3> | + | <!--navbar--> |
− | <p>This page is used by the judges to evaluate your team for the <a href="https://2021.igem.org/Judging/Medals">medal criterion</a> or <a href="https://2021.igem.org/Judging/Awards"> award listed below</a>. </p> | + | <div class="navbar"> |
− | <p> Delete this box in order to be evaluated for this medal criterion and/or award. See more information at <a href="https://2021.igem.org/Judging/Pages_for_Awards"> Instructions for Pages for awards</a>.</p>
| + | <a class="logo" href="https://2021.igem.org/Team:Shanghai_Metropolis"> |
− | </div>
| + | <img src="https://static.igem.org/mediawiki/2021/7/72/T--Shanghai_Metropolis--logo.png" alt=""> |
| + | </a> |
| | | |
| + | <!--nav--> |
| + | <ul class="list"> |
| + | <!--Team--> |
| + | <li class="item"> |
| + | <a href="https://2021.igem.org/Team:Shanghai_Metropolis/Team">Team</a> |
| + | <div class="child-nav"> |
| + | <ul class="child-list"> |
| + | <li class="child-item"> |
| + | <a href="https://2021.igem.org/Team:Shanghai_Metropolis/Team">Team Members</a> |
| + | </li> |
| + | <li class="child-item"> |
| + | <a href="https://2021.igem.org/Team:Shanghai_Metropolis/Attributions">Attributions</a> |
| + | </li> |
| + | <li class="child-item"> |
| + | <a href="https://2021.igem.org/Team:Shanghai_Metropolis/Collaborations">Collaboration</a> |
| + | </li> |
| + | <li class="child-item"> |
| + | <a href="https://2021.igem.org/Team:Shanghai_Metropolis/Partnership">Partnership</a> |
| + | </li> |
| + | </ul> |
| + | </div> |
| + | </li> |
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− | <div class="clear"></div> | + | <!--Modeling--> |
| + | <li class="item"> |
| + | <a href="https://2021.igem.org/Team:Shanghai_Metropolis/Model">Modeling</a> |
| + | </li> |
| | | |
| + | <!--Entrepreneurship--> |
| + | <li class="item"> |
| + | <a href="https://2021.igem.org/Team:Shanghai_Metropolis/Entrepreneurship">Entrepreneurship</a> |
| + | </li> |
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− | <div class="column full_size"> | + | <!--Implementation--> |
− | <h1>Engineering Success</h1> | + | <li class="item"> |
− | <h3> Silver Medal Criterion #1</h3>
| + | <a href="https://2021.igem.org/Team:Shanghai_Metropolis/Implementation">Implementation</a> |
| + | </li> |
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− | <p>Demonstrate engineering success in a part of your project by going through at least one iteration of the engineering design cycle. This achievement should be distinct from your Contribution for Bronze. | + | <!--Human Practice--> |
− | <br><br> | + | <li class="item"> |
− | Please see the <a href="https://2021.igem.org/Judging/Medals">2021 Medals Page</a> for more information.
| + | <a href="https://2021.igem.org/Team:Shanghai_Metropolis/Human_Practices">Human Practice</a> |
− | </p> | + | <div class="child-nav"> |
| + | <ul class="child-list"> |
| + | <li class="child-item"> |
| + | <a href="https://2021.igem.org/Team:Shanghai_Metropolis/Human_Practices">Integrated Human |
| + | Practice</a> |
| + | </li> |
| + | <li class="child-item"> |
| + | <a href="https://2021.igem.org/Team:Shanghai_Metropolis/Communication">Communication</a> |
| + | </li> |
| + | <li class="child-item"> |
| + | <a href="https://2021.igem.org/Team:Shanghai_Metropolis/Fundraising">Fundraising</a> |
| + | </li> |
| + | </ul> |
| + | </div> |
| + | </li> |
| | | |
| + | <!--Parts--> |
| + | <li class="item active"> |
| + | <a href="https://2021.igem.org/Team:Shanghai_Metropolis/PartsCollection">Parts</a> |
| + | <div class="child-nav"> |
| + | <ul class="child-list"> |
| + | <li class="child-item"> |
| + | <a href="https://2021.igem.org/Team:Shanghai_Metropolis/Parts_Collection">Parts |
| + | Collection</a> |
| + | </li> |
| + | <li class="child-item active"> |
| + | <a href="https://2021.igem.org/Team:Shanghai_Metropolis/Engineering">Engineering</a> |
| + | </li> |
| + | </ul> |
| + | </div> |
| + | </li> |
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− | <p>If you plan to show engineering success by creating a new Part that has been shown to work as expected, you must document your contribution on the Part's Main Page on the <a href="http://parts.igem.org/Main_Page">Registry</a> for your team to be eligible for this criteria.</p> | + | <!--Project--> |
| + | <li class="item"> |
| + | <a href="https://2021.igem.org/Team:Shanghai_Metropolis/Description">Project</a> |
| + | <div class="child-nav"> |
| + | <ul class="child-list"> |
| + | <li class="child-item"> |
| + | <a href="https://2021.igem.org/Team:Shanghai_Metropolis/Description">Description</a> |
| + | </li> |
| + | <li class="child-item"> |
| + | <a href="https://2021.igem.org/Team:Shanghai_Metropolis/Experiments">Experiments</a> |
| + | </li> |
| + | <li class="child-item"> |
| + | <a href="https://2021.igem.org/Team:Shanghai_Metropolis/Results">Result</a> |
| + | </li> |
| + | <li class="child-item"> |
| + | <a href="https://2021.igem.org/Team:Shanghai_Metropolis/Proof_Of_Concept">Proof of Concept</a> |
| + | </li> |
| + | <li class="child-item"> |
| + | <a href="https://2021.igem.org/Team:Shanghai_Metropolis/Notebook">Notebook</a> |
| + | </li> |
| + | <li class="child-item"> |
| + | <a href="https://2021.igem.org/Team:Shanghai_Metropolis/Safety">Safety</a> |
| + | </li> |
| + | </ul> |
| + | </div> |
| + | </li> |
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− | </div> | + | <!--Home--> |
| + | <li class="item"> |
| + | <a href="https://2021.igem.org/Team:Shanghai_Metropolis">Home</a> |
| + | </li> |
| + | </ul> |
| + | </div> |
| | | |
| + | <!--图片--> |
| + | <div class="page-header"> |
| + | <img src="https://static.igem.org/mediawiki/2021/d/d2/T--Shanghai_Metropolis--bg-1.png" alt=""> |
| + | <h1>Engineering</h1> |
| + | </div> |
| | | |
| + | <!--内容--> |
| + | <div class="content-small"> |
| + | <section class="article p-t-54 p-b-54"> |
| + | <section> |
| + | <h1 class="title">Introduction </h1> |
| + | |
| + | <p>Hand-foot-mouth disease (HFMD) is an infectious disease caused by enterovirus 71 (EV71). The virus is an |
| + | important pathogenic factor of hand, foot and mouth disease. Vp1 protein is the viral capsid protein and |
| + | promotes the infection of host cells by virus particles. Vp1 is also the main antigen gene of the EV71 |
| + | virus. Generally, the vaccinated population, especially infants and young children, are more compliant |
| + | with oral vaccines, so we are trying to develop oral HFMD vaccines. Probiotics bacteria <i>Bifidobacteria</i>, |
| + | as the natural host of the intestinal tract, can adhere to intestinal epithelial cells and are ideal |
| + | oral live vaccine expression vectors, and related studies have found that their preventive effects on |
| + | gastrointestinal pathogens are more significant. Therefore, we can use the <i>bifidobacterium</i> in |
| + | lactic acid bacteria as an expression system to express EV71 vp1.</p> |
| + | </section> |
| + | |
| + | <section> |
| + | <h1 class="title">Design </h1> |
| + | |
| + | <p>The B subunit LTB in the heat-labile enterotoxin (LT) of Escherichia coli heat-labile enterotoxin (LT) |
| + | has strong immunogenicity and adjuvant activity, and will not cause harm to the human body. LTB and a |
| + | variety of non-related proteins and their non-protein antigens can increase the mucosal IgA and humoral |
| + | immune IgG response levels of the antigen through different immunization pathways. Fusion recombinant |
| + | protein constituting EV71 vp1 and LTB is produced by bifidobacterium expression system. Vp1-LTB is |
| + | tagged by glutathione S-transferase (GST) in its N-terminal. The tag is used for recombinant protein |
| + | purification. </p> |
| + | </section> |
| + | |
| + | <section> |
| + | <h1 class="title">Build </h1> |
| + | |
| + | <p>Plasmids pGEX-vp1 and pGEX-vp1-LTB were constructed to produce Vp1 and Vp1-LTB fusion proteins in |
| + | probiotics bacteria Bifidobacterium, respectively (Figure 1). The production is controlled by tac |
| + | promotor. The construct is confirmed by DNA sequencing.</p> |
| + | |
| + | <div class="img-container"> |
| + | <img src="https://static.igem.org/mediawiki/2021/f/fd/T--Shanghai_Metropolis--img_engineering_1.jpg" alt="" style="width: 80%"> |
| + | <span class="figure">Figure 1. Schematic map of plasmids pGEX-vp and pGEX-vp1-LTB.</span> |
| + | </div> |
| + | </section> |
| + | |
| + | <section> |
| + | <h1 class="title">Test </h1> |
| + | |
| + | <p>Due to the relatively low rate of growth and efficiency of electroporation of <i>L. casei</i>, our team |
| + | first transformed <i>E. coli</i> BL21, which is commonly used in plasmids transformation, to verify the |
| + | expression and antigencity of VP1 and VP1-LTB proteins (Figure 2).</p> |
| + | |
| + | <div class="img-container"> |
| + | <img src="https://static.igem.org/mediawiki/2021/a/a2/T--Shanghai_Metropolis--img_engineering_2.jpg" alt="" style="width: 70%"> |
| + | <span class="figure">Figure 2. SDS-PAGE and Western Blot for expression of VP1 and VP1-LTB proteins.</span> |
| + | </div> |
| + | </section> |
| + | |
| + | <section> |
| + | <h1 class="title">Learn</h1> |
| + | |
| + | <p>GST, GST-VP1 and GST-VP1-LTB had all been successfully expressed by E.coli BL21, and they mainly existed |
| + | in the precipitation in the form of inclusion body. The expression of VP1 and VP1-LTB was not so |
| + | effective in <i>E.coli</i> BL21. There was no obvious expression of GST-VP1 and GST-VP1-LTB in <i>L. |
| + | casei</i>.</p> |
| + | </section> |
| + | </section> |
| + | </div> |
| + | |
| + | <!--Footer--> |
| + | <div class="footer"> |
| + | <div class="content"> |
| + | <hr> |
| + | <span>Email us at: zhixuansong1022@qq.com; 1956461994@qq.com</span> |
| + | <span>Search‘EV71terminator’ in Wechat to find more about us</span> |
| + | </div> |
| + | </div> |
| + | </div> |
| </html> | | </html> |