Difference between revisions of "Team:HUST-China/HP/Silver"

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{{HUST-China}}
 
 
<html>
 
<html>
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<div class="column full_size judges-will-not-evaluate">
 
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<h3>★  ALERT! </h3>
 
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<p>This page is used by the judges to evaluate your team for the <a href="https://2017.igem.org/Judging/Medals">medal criterion</a> or <a href="https://2017.igem.org/Judging/Awards"> award listed above</a>. </p>
 
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<p> Delete this box in order to be evaluated for this medal criterion and/or award. See more information at <a href="https://2017.igem.org/Judging/Pages_for_Awards"> Instructions for Pages for awards</a>.</p>
 
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</div>
 
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<div class="clear"></div>
 
  
 +
<head>
 +
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 +
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 +
    <meta name="renderer" content="webkit">
 +
    <title>Experiment</title>
 +
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 +
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    <script type="text/javascript" src="https://2017.igem.org/Team:HUST-China/js/jquery?action=raw&amp;ctype=text/javascript"></script>
 +
    <link href="https://2017.igem.org/Team:HUST-China/css/return_top?action=raw&amp;ctype=text/css" rel="stylesheet" type="text/css" />
 +
    <script type="text/javascript" src="https://2017.igem.org/Team:HUST-China/js/return_top_jquery?action=raw&amp;ctype=text/javascript"></script>
 +
    <script src="https://2017.igem.org/Team:HUST-China/js/return_top_script?action=raw&amp;ctype=text/javascript" type="text/javascript"></script>
  
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<div class="column full_size">
+
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    @media (min-width: 1200px) {
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            /* margin-left: 300px; */
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 +
            width: 1170px;
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    /* article */
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        margin: 10px 0;
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    .text-content article img {
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        padding-left: 20%;
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        /* margin-left: 20%; */
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        margin-left: -30%;
 +
        text-align: center;
 +
        list-style: none;
 +
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<h1>Silver Medal Human Practices</h1>
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<p>iGEM teams are leading in the area of Human Practices because they conduct their projects within a social/environmental context, to better understand issues that might influence the design and use of their technologies.</p>
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        @charset "utf-8";
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<p>Teams work with students and advisors from the humanities and social sciences to explore topics concerning ethical, legal, social, economic, safety or security issues related to their work. Consideration of these Human Practices is crucial for building safe and sustainable projects that serve the public interest. </p>
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    body {
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<p>For more information, please see the <a href="https://2017.igem.org/Competition/Human_Practices">Human Practices page</a>.</p>
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        font-family: "Helvetica Neue", Helvetica, Arial, "Microsoft Yahei UI", "Microsoft YaHei", SimHei, "\5B8B\4F53", simsun, sans-serif;
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</div>
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 +
        width:100%;
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    }
 +
    .logo {
 +
        padding:0;
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    }
  
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<div class="clear"></div>
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    #myCarousel {
 +
        margin: 50px 0 0 0;
 +
    }
 +
    #navbar-collapse ul {
 +
        margin-top:0;
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    .carousel-inner img {
 +
        margin: 0 auto;
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 +
        font-size: 100px;
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 +
        color: #ffffff;
 +
        background-color:#97477c;
 +
        padding: 30px;
 +
        margin: 0;
 +
    }
  
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<div class="column half_size">
+
    .tab-p {
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<h3>Silver Medal Criterion #3</h3>
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        font-size: 20px;
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<p>Convince the judges you have thought carefully and creatively about whether your work is safe, responsible and good for the world. You could accomplish this through engaging with your local, national and/or international communities or other approaches. Please note that standard surveys will not fulfill this criteria.</p>
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        text-align: center;
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</div>
+
        letter-spacing: 1px;
 +
        color: #999;
 +
        margin: 20px 0 40px 0;
 +
    }
  
−
<div class="column half_size">
+
    .container-fluid{
−
<h5>Some Human Practices topic areas </h5>
+
        width:100%;
−
<ul>
+
    }
−
<li>Philosophy</li>
+
−
<li>Public Engagement / Dialogue</li>
+
−
<li>Education</li>
+
−
<li>Product Design</li>
+
−
<li>Scale-Up and Deployment Issues</li>
+
−
<li>Environmental Impact</li>
+
−
<li>Ethics</li>
+
−
<li>Safety</li>
+
−
<li>Security</li>
+
−
<li>Public Policy</li>
+
−
<li>Law and Regulation</li>
+
−
<li>Risk Assessment</li>
+
−
</ul>
+
−
</div>
+
  
 +
    .tab1 {
 +
        width:100%;
 +
        margin: 0 0;
 +
        color: #666;
 +
        background:#333;
 +
    }
  
−
<div class="column half_size">
+
    .tab1 .text-muted {
−
<h5>What should we write about on this page?</h5>
+
        color: #999;
−
<p>On this page, you should write about the Human Practices topics you considered in your project, and document any special activities you did (such as visiting experts, talking to lawmakers, or doing public engagement). This should include all of the work done for the Silver Medal Criterion #3. Details for your Gold medal work and/or work for the two Human Practices special prizes should be put on those specified pages.</p>
+
        text-decoration: line-through;
−
</div>
+
    }
 +
    .tab1 .media-heading {
 +
        margin: 5px 0 20px 0;
 +
    }
 +
    .tab1 .col {
 +
        padding: 20px;
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 +
    .text span{
 +
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 +
    .text p {
 +
        font-size: 14px;
 +
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 +
    #footer {
 +
        background-color: #eee;
 +
        border-top: 1px solid #ccc;
 +
        padding: 20px;
 +
        text-align: center;
 +
    }
 +
    .text h4{
 +
        padding: 5px 10px;
 +
    }
 +
    .text p{
 +
        padding: 2px 10px;
 +
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 +
    /*没有小于768px，是因为Bootstrap3以移动端优先设计*/
  
−
<div class="column half_size">
+
    /* 小屏幕（平板，大于等于768px） */
−
<h5>Inspiration</h5>
+
    @media (min-width: 768px) {
−
<p>Read what other teams have done:</p>
+
        .tab-h1 {
−
<ul>
+
            font-size: 26px;
−
<li><a href="https://2014.igem.org/Team:Dundee/policypractice/experts">2014 Dundee </a></li>
+
        }
−
<li><a href="https://2014.igem.org/Team:UC_Davis/Policy_Practices_Overview">2014 UC Davis </a></li>
+
        .tab-p {
−
<li><a href="https://2013.igem.org/Team:Manchester/HumanPractices">2013 Manchester </a></li>
+
            font-size: 16px;
−
<li><a href="https://2013.igem.org/Team:Cornell/outreach">2013 Cornell </a></li>
+
        }
−
</ul>
+
        .text h3 {
−
</div>
+
            font-size: 22px;
 +
        }
 +
        .text p {
 +
            font-size: 10px;
 +
            text-indent: 20px;
 +
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 +
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 +
            float: left;
 +
        }
 +
        .tab2-img {
 +
            float: right;
 +
        }
 +
        .experiment_circuit img{
 +
        width:80%;
 +
        height:auto;
 +
        }
 +
    }
  
 +
    /* 中等屏幕（桌面显示器，大于等于992px） */
 +
    @media (min-width: 992px) {
 +
        .tab-h1 {
 +
            font-size: 28px;
 +
        }
 +
        .tab-p {
 +
            font-size: 17px;
 +
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 +
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 +
            font-size: 24px;
 +
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 +
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 +
          font-size: 12px;
 +
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 +
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 +
        width:60%;
 +
        height:auto;
 +
        }
 +
    }
  
 +
    /* 大屏幕（大桌面显示器，大于等于1200px） */
 +
    @media (min-width: 1200px) {
 +
        .tab-h1 {
 +
            font-size: 30px;
 +
        }
 +
        .tab-p {
 +
            font-size: 18px;
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 +
            text-indent: 28px;
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 +
        width:40%;
 +
        height:auto;
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 +
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 +
       
 +
 +
    body {
 +
            position: relative;
 +
        }
 +
        ul.nav-pills {
 +
            top: 200px;
 +
            position: fixed;
 +
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 +
        div.col-sm-10 div {
 +
            min-height: 100px;
 +
            font-size: 14px;
 +
            text-indent: 28px;
 +
            color:#0f0f0f;
 +
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 +
 +
        @media screen and (max-width: 810px) {
 +
            #section1, #section2, #section3, #section4, #section5 {
 +
            background-color: #eee; color:#171717;
 +
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 +
        }
 +
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    </style>
 +
</head>
 +
 +
<body data-spy="scroll" data-target="#myScrollspy" data-offset="20">
 +
      <nav class="navbar navbar-default navbar-fixed-top" style="margin-top:15px">
 +
        <div class="container">
 +
 +
            <div class="navbar-header">
 +
                <a href="https://2017.igem.org/Team:HUST-China" class="navbar-brand logo" style="padding:0 5px;"><img src="https://static.igem.org/mediawiki/2017/6/64/Team_HUST_China_2017_logo.png" alt="HUST_iGEM_2017_logo"></a>
 +
                <a href="https://2017.igem.org" class="navbar-brand logo" style="padding:0 5px;"><img src="https://static.igem.org/mediawiki/2017/f/f6/IGEM_2017_logo.png" alt="HUST_iGEM_2017_logo"></a>
 +
                <button type="button" class="navbar-toggle" data-toggle="collapse" data-target="#navbar-collapse">
 +
                    <span class="glyphicon glyphicon-list"></span>
 +
                </button>
 +
            </div>
 +
 +
 +
            <div class="collapse navbar-collapse" id="navbar-collapse">
 +
                <ul class="nav navbar-nav navbar-right">
 +
 +
 +
 +
                    <li><a href="https://2017.igem.org/Team:HUST-China">HOME</a></li>
 +
                    <li class="dropdown">
 +
                        <a href="#" class="dropdown-toggle" data-toggle="dropdown">
 +
                            PROJECT <b class="caret"></b>
 +
                        </a>
 +
                        <ul class="dropdown-menu">
 +
                            <li><a href="https://2017.igem.org/Team:HUST-China/project/background">Background</a></li>
 +
                            <li><a href="https://2017.igem.org/Team:HUST-China/Description">Description</a></li>                                       
 +
                            <li><a href="https://2017.igem.org/Team:HUST-China/Safety">Safety</a></li>
 +
                        </ul>
 +
                    </li>
 +
                    <li class="dropdown active">
 +
                        <a href="#" class="dropdown-toggle" data-toggle="dropdown">
 +
                            WETLAB <b class="caret"></b>
 +
                        </a>
 +
                        <ul class="dropdown-menu">
 +
                            <li><a href="https://2017.igem.org/Team:HUST-China/Experiments">Experiments</a></li>
 +
                            <li><a href="https://2017.igem.org/Team:HUST-China/Results">Results</a></li>
 +
                            <li><a href="https://2017.igem.org/Team:HUST-China/InterLab">Interlab</a></li>
 +
                            <li><a href="https://2017.igem.org/Team:HUST-China/Notebook">Notebook</a></li>
 +
                            <li><a href="https://2017.igem.org/Team:HUST-China/wetlab/protocols">Protocols</a></li>
 +
                        </ul>
 +
                    </li>
 +
                    <li class="dropdown">
 +
                        <a href="https://2017.igem.org/Team:HUST-China/Modeling" >
 +
                            MODELING
 +
                        </a>
 +
                    </li>
 +
                    <li class="dropdown">
 +
                        <a href="https://2017.igem.org/Team:HUST-China/Parts">
 +
                            PARTS
 +
                        </a>
 +
                    </li>
 +
                    <li class="dropdown">
 +
                        <a href="https://2017.igem.org/Team:HUST-China/Human_Practices">
 +
                            HUMAN PRACTICES
 +
                        </a>
 +
                    </li>
 +
                    <li class="dropdown">
 +
                        <a href="#" class="dropdown-toggle" data-toggle="dropdown">
 +
                            TEAM <b class="caret"></b>
 +
                        </a>
 +
                        <ul class="dropdown-menu">
 +
                            <li><a href="https://2017.igem.org/Team:HUST-China/Team">Team Roster</a></li>
 +
                            <li><a href="https://2017.igem.org/Team:HUST-China/Attributions">Attributions</a></li>
 +
                        </ul>
 +
                    </li>
 +
 +
 +
                </ul> 
 +
            </div>
 +
        </div>
 +
    </nav>
 +
 +
 +
    <div class="tab1" style="width:100%;margin-top: 65px;">
 +
            <h1 class="tab-h1" ><strong>「Experiment」</strong></h1>
 +
    </div>
 +
 +
    <div class="container-fluid text">
 +
        <div class="row">
 +
            <nav class="col-sm-2" id="myScrollspy">
 +
                <div class="container-fluid">
 +
                <div class="collapse navbar-collapse" id="navbar-collapse">
 +
                <ul class="nav nav-pills nav-stacked" style="font-size:12px; text-align:center; background:#ccc;">
 +
                    <li class="active"><a href="#section1">Sensing part</a></li>
 +
                    <li><a href="#section2">Recycling part</a></li>
 +
                    <li><a href="#section3">The whole circuit </a></li>
 +
                </ul>
 +
                </div> 
 +
                </div>     
 +
            </nav>
 +
            <div class="col-sm-8">
 +
                <div id="section1" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;">   
 +
                    <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px;margin-top:0;">Sensing part</h3>
 +
                    <p>
 +
                        The sensing part is to achieve the awareness of the engineering bacteria on the rare earth ions in the environment.
 +
                    </p>
 +
                    <h4><strong>①Amplify</strong> </h4>
 +
                    <p>
 +
                        pmrA-B-Fe、pmrC/GFP  gene through the pcr protocol
 +
                    </p>
 +
                    <h4><strong>②Plasmid Construction</strong> </h4>
 +
                    <p>
 +
                      Coding sequences pmrA/pmrB and pmrC-GFP were cloned into the expression vector pBAD30 by restriction enzyme digestion and DNA ligation.
 +
                    </p>
 +
                    <div class="container experiment_circuit col-md-6 col-xs-6 col-md-offset-3 col-xs-offset-3">
 +
                        <img src="https://static.igem.org/mediawiki/2017/d/d5/2017_HUST_China_Experiment_image1.png" alt="experiment_circuit" class="img-responsive">
 +
                    </div>
 +
                    <div class="row col-xs-12">
 +
                        <p>
 +
                            Then we transformed these vectors into Ecoli BL21 cells.
 +
                        </p>
 +
                        <h4><strong>③Detection</strong> </h4>
 +
                        <p>
 +
                          Arabinose was used to induce expression of pmrABC system.
 +
                        </p>
 +
                        <p>
 +
                          We use the original pmrA-pmrB / Fe-pmrC-GFP circuit to test whether our
 +
                          pmrABC system works in our plasmid, then the expression and efficacy
 +
                          of LBT1-12 were tested .
 +
                        </p>
 +
                    </div>
 +
 +
 +
                    <div class="table-responsive" style="padding: 10px 100px; text-align: center; font-size:14px;">
 +
                        <table class="table">
 +
                          <thead>
 +
                              <tr>
 +
                                <th style="text-align: center;">gene circuit</th>
 +
                                <th style="text-align: center;">vector</th>
 +
                                <th style="text-align: center;">description</th>
 +
                              </tr>
 +
                          </thead>
 +
                          <tbody>
 +
                                  <tr>
 +
                                    <td>pmrA-pmrB/Fe-pmrC-GFP</td>
 +
                                    <td>pBAD30</td>
 +
                                    <td>Test whether our pmrABC system works in our plasmid</td>
 +
                                  </tr>
 +
                                  <tr>
 +
                                    <td>pmrA-pmrB/LBT1-pmrC-GFP </td>
 +
                                    <td>pBAD30</td>
 +
                                    <td> Detecting effectiveness of protein LBT1</td>
 +
                                  </tr>
 +
                                  <tr>
 +
                                    <td>pmrA-pmrB/LBT2-pmrC-GFP</td>
 +
                                    <td>pBAD30 </td>
 +
                                    <td> Detecting effectiveness of protein LBT2</td>
 +
                                  </tr>
 +
                                  <tr>
 +
                                    <td>pmrA-pmrB/LBT3-pmrC-GFP</td>
 +
                                    <td>pBAD30 </td>
 +
                                    <td>Detecting effectiveness of protein LBT3</td>
 +
                                  </tr>
 +
                                  <tr>
 +
                                    <td>pmrA-pmrB/LBT4-pmrC-GFP</td>
 +
                                    <td> pBAD30 </td>
 +
                                    <td>Detecting effectiveness of protein LBT4</td>
 +
                                  </tr>
 +
                                  <tr>
 +
                                    <td>pmrA-pmrB/LBT5-pmrC-GFP</td>
 +
                                    <td>pBAD30</td>
 +
                                    <td>Detecting effectiveness of protein LBT5</td>
 +
                                  </tr>
 +
                                  <tr>
 +
                                    <td>pmrA-pmrB/LBT6-pmrC-GFP </td>
 +
                                    <td>pBAD30</td>
 +
                                    <td> Detecting effectiveness of protein LBT6</td>
 +
                                  </tr>
 +
                                  <tr>
 +
                                    <td>pmrA-pmrB/LBT7-pmrC-GFP</td>
 +
                                    <td>pBAD30 </td>
 +
                                    <td> Detecting effectiveness of protein LBT7</td>
 +
                                  </tr>
 +
                                  <tr>
 +
                                    <td>pmrA-pmrB/LBT8-pmrC-GFP</td>
 +
                                    <td>pBAD30 </td>
 +
                                    <td>Detecting effectiveness of protein LBT8</td>
 +
                                  </tr>
 +
                                  <tr>
 +
                                    <td>pmrA-pmrB/LBT9-pmrC-GFP</td>
 +
                                    <td> pBAD30 </td>
 +
                                    <td>Detecting effectiveness of protein LBT9</td>
 +
                                  </tr>
 +
                                  <tr>
 +
                                    <td>pmrA-pmrB/LBT10-pmrC-GFP</td>
 +
                                    <td>pBAD30 </td>
 +
                                    <td> Detecting effectiveness of protein LBT10</td>
 +
                                  </tr>
 +
                                  <tr>
 +
                                    <td>pmrA-pmrB/LBT11-pmrC-GFP</td>
 +
                                    <td>pBAD30 </td>
 +
                                    <td>Detecting effectiveness of protein LBT11</td>
 +
                                  </tr>
 +
                                  <tr>
 +
                                    <td>pmrA-pmrB/LBT12-pmrC-GFP </td>
 +
                                    <td> pBAD30 </td>
 +
                                    <td>Detecting effectiveness of protein LBT12</td>
 +
                                  </tr>
 +
                               
 +
                          </tbody>
 +
                        </table>
 +
                    </div>
 +
                </div>
 +
 +
 +
                <div id="section2" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;">
 +
                    <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">Recycling part </h3>
 +
                    <p>
 +
                            The recycling part is to capture the rare earth ions in the environment and to anchor our cells over the silicon net.
 +
                    </p>
 +
                    <h4><strong>①With appropriate primers, PCR was carried out to amplify target gene oprf-LBT1-12 and oprf-Si tag.</strong> </h4>
 +
                    <div class="container experiment_circuit col-md-6 col-xs-6 col-md-offset-3 col-xs-offset-3">
 +
                        <img src="https://static.igem.org/mediawiki/2017/7/73/2017_HUST_China_Experiment_image2.png" alt="experiment_circuit" class="img-responsive">
 +
                    </div>
 +
                    <h4 class="col-xs-12"><strong>②Then we constructed two vectors to achieve expression of LBT and Si-tag.</strong> </h4>
 +
                    <div class="container experiment_circuit col-md-6 col-xs-6 col-md-offset-3 col-xs-offset-3">
 +
                        <img src="https://static.igem.org/mediawiki/2017/7/78/2017_HUST_China_Experiment_image3.png" alt="experiment_circuit" class="img-responsive">
 +
                    </div>
 +
                    <p class="col-xs-12">
 +
                        We cloned the sequence of oprf-LBT and Si-tag into the vector pET28α in order to make the sequence be tagged N-terminally with a 6×His tag. 6×His tag is an affinity tag which allows the tagged recombinant protein to be purified in the process of affinity purification.
 +
                    </p>
 +
                    <h4><strong>③Detection</strong> </h4>
 +
                    <p>
 +
                      In the meanwhile,we added the FLAG tag to achieve that whether the LBTs and the Si tag was expressed on the cell membrane.
 +
                    </p>
 +
                </div>
 +
 +
                <div id="section3" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;">       
 +
                    <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">Construction of the whole circuit</h3>
 +
                    <p>
 +
                      All of our part should be built in one plasmid. Therefore, we use endonuclease and ligase to  replace the GFP by Oprf-LBT and Oprf-Si tag.
 +
                    </p>
 +
                    <div class="container experiment_circuit col-md-6 col-xs-6 col-md-offset-3 col-xs-offset-3" style="padding: 5px;">
 +
                        <img src="https://static.igem.org/mediawiki/2017/5/58/2017_HUST_China_Experiment_image4.png" alt="experiment_circuit" class="img-responsive">
 +
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Revision as of 13:38, 27 October 2017

Experiment

「Experiment」

Sensing part

The sensing part is to achieve the awareness of the engineering bacteria on the rare earth ions in the environment.

①Amplify

pmrA-B-Fe、pmrC/GFP gene through the pcr protocol

②Plasmid Construction

Coding sequences pmrA/pmrB and pmrC-GFP were cloned into the expression vector pBAD30 by restriction enzyme digestion and DNA ligation.

experiment_circuit

Then we transformed these vectors into Ecoli BL21 cells.

③Detection

Arabinose was used to induce expression of pmrABC system.

We use the original pmrA-pmrB / Fe-pmrC-GFP circuit to test whether our pmrABC system works in our plasmid, then the expression and efficacy of LBT1-12 were tested .

gene circuit vector description
pmrA-pmrB/Fe-pmrC-GFP pBAD30 Test whether our pmrABC system works in our plasmid
pmrA-pmrB/LBT1-pmrC-GFP pBAD30 Detecting effectiveness of protein LBT1
pmrA-pmrB/LBT2-pmrC-GFP pBAD30 Detecting effectiveness of protein LBT2
pmrA-pmrB/LBT3-pmrC-GFP pBAD30 Detecting effectiveness of protein LBT3
pmrA-pmrB/LBT4-pmrC-GFP pBAD30 Detecting effectiveness of protein LBT4
pmrA-pmrB/LBT5-pmrC-GFP pBAD30 Detecting effectiveness of protein LBT5
pmrA-pmrB/LBT6-pmrC-GFP pBAD30 Detecting effectiveness of protein LBT6
pmrA-pmrB/LBT7-pmrC-GFP pBAD30 Detecting effectiveness of protein LBT7
pmrA-pmrB/LBT8-pmrC-GFP pBAD30 Detecting effectiveness of protein LBT8
pmrA-pmrB/LBT9-pmrC-GFP pBAD30 Detecting effectiveness of protein LBT9
pmrA-pmrB/LBT10-pmrC-GFP pBAD30 Detecting effectiveness of protein LBT10
pmrA-pmrB/LBT11-pmrC-GFP pBAD30 Detecting effectiveness of protein LBT11
pmrA-pmrB/LBT12-pmrC-GFP pBAD30 Detecting effectiveness of protein LBT12

Recycling part

The recycling part is to capture the rare earth ions in the environment and to anchor our cells over the silicon net.

①With appropriate primers, PCR was carried out to amplify target gene oprf-LBT1-12 and oprf-Si tag.

experiment_circuit

②Then we constructed two vectors to achieve expression of LBT and Si-tag.

experiment_circuit

We cloned the sequence of oprf-LBT and Si-tag into the vector pET28α in order to make the sequence be tagged N-terminally with a 6×His tag. 6×His tag is an affinity tag which allows the tagged recombinant protein to be purified in the process of affinity purification.

③Detection

In the meanwhile,we added the FLAG tag to achieve that whether the LBTs and the Si tag was expressed on the cell membrane.

Construction of the whole circuit

All of our part should be built in one plasmid. Therefore, we use endonuclease and ligase to replace the GFP by Oprf-LBT and Oprf-Si tag.

experiment_circuit