Difference between revisions of "Team:XMU-China/Proof Of Concept"

Line 1: Line 1:
−
{{IGEM_TopBar}}
+
<html lang="en">
−
{{XMU-China}}
+
<head>
−
<html>
+
    <meta charset="UTF-8" name="viewport" content="width=device-width, initial-scale=1.0, maximum-scale=1.0, user-scalable=0">
 +
    <meta http-equiv="X-UA-Compatible" content="IE=edge,chrome=1">
 +
    <meta name="viewport" content="width=500" />
 +
    <meta name="keywords" content="ScrollMagic, example, scrolling, attaching, scrollbar, tween, tweenmax" />
 +
    <meta name="author" content="Jan Paepke (www.janpaepke.de)" />
 +
    <meta name="viewport" content="width=device-width,initial-scale=1.0,maximum-scale=1.0,minimum-scale=1.0,user-scalable=no">
 +
    <style type='text/css'>
 +
        #sideMenu, #top_title, .patrollink, #firstHeading, #home_logo, #sideMenu { display:none; }
 +
        #content { padding:0px; width:100%; margin-top:-7px; margin-left:0px; border:none;}
 +
        body, html {background-color:white; width: 100%; height: 100%;}
 +
        #bodyContent h1, #bodyContent h2, #bodyContent h3, #bodyContent h4, #bodyContent h5 { margin-bottom: 0px; }
 +
        #bodyContent a[href ^="https://"], .link-https { padding-right:0px;}
 +
        #home_logo, #sideMenu { display:none; }
 +
    </style>
 +
    <link rel="stylesheet" href="https://2020.igem.org/Team:XMU-China/css/initialize/css?action=raw&amp;ctype=text/css">
 +
    <link rel="stylesheet" href="https://2020.igem.org/Team:XMU-China/css/part1listpoc?action=raw&amp;ctype=text/css">
 +
    <link rel="stylesheet" href="https://2020.igem.org/Team:XMU-China/css/universial?action=raw&amp;ctype=text/css">
 +
    <link rel="stylesheet" href="https://2020.igem.org/Team:XMU-China/css/5menu?action=raw&amp;ctype=text/css">
 +
    <link rel="stylesheet" href="https://2020.igem.org/Team:XMU-China/css/rightnav?action=raw&amp;ctype=text/css">
 +
    <link href="https://cdn.bootcss.com/font-awesome/4.7.0/css/font-awesome.min.css" rel="stylesheet">
 +
    <script src="https://2020.igem.org/Team:XMU-China/js/jquery?action=raw&amp;ctype=text/javascript"></script>
 +
    <script src="https://2020.igem.org/Team:XMU-China/js/bootstrap?action=raw&amp;ctype=text/javascript"></script>
 +
</head>
 +
<nav class="Quick-navigation">
 +
    <div class="Quick-navigation_word">
 +
        <img src="https://static.igem.org/mediawiki/2020/e/eb/T--XMU-China--right0-2.png">
 +
        <a href="#P0" class="Quick-navigation-item">
 +
            <img id="turn_img" src="https://static.igem.org/mediawiki/2020/6/62/T--XMU-China--right1-2.png">
 +
            <a href="#P0"id="Quick_A">Kill switch</a></a>
 +
        <a href="#P1" class="Quick-navigation-item" >
 +
            <img id="turn_img" src="https://static.igem.org/mediawiki/2020/d/dd/T--XMU-China--right2-2.png">
 +
            <a href="#P1" id="Quick_B">The kill effect</a></a>
 +
        <a href="#Safe_Shipment" class="Quick-navigation-item">
 +
            <img id="turn_img" src="https://static.igem.org/mediawiki/2020/d/dd/T--XMU-China--right2-2.png">
 +
            <a href="#Safe_Shipment" id="Quick_C"></a></a>
 +
    </div>
 +
</nav>
 +
<body>
 +
<div class="containor">
 +
    <div class="part1">
 +
        <div class="wrap">
 +
            <div class="team">
 +
                <a href="https://2020.igem.org/Team:XMU-China">
 +
                    <img src="https://static.igem.org/mediawiki/2020/c/cb/T--XMU-China--logo.png" alt="">
 +
                </a>
 +
            </div>
 +
            <div class="aaa">
 +
                <ul class="level">
 +
                    <li>
 +
                        <div class="ball"></div><a href="https://2020.igem.org/Team:XMU-China" class="a1">Home</a>
 +
                    </li>
 +
                    <li>
 +
                        <div class="ball"></div>
 +
                        <a href="" class="a1">Team</a>
 +
                        <ul class="two">
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Team"><li>Members</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Collaborations"><li>Collaborations</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Partnership"><li>Partnership</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Attributions"><li>Attributions</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Notebook"><li>Notebook</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Experiments"><li>Experiments</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Safety"><li>Safety</li></a>
 +
                        </ul>
 +
                    </li>
 +
                    <li>
 +
                        <div class="ball"></div>
 +
                        <a href="" class="a1">Project</a>
 +
                        <ul class="two">
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Description"><li>Description</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Design"><li>Design</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Model"><li>Models</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Results"><li>Result</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Proof_Of_Concept"><li>Proof of Concept</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Hardware"><li>Hardware</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Implementation"><li>Proposed Implementation</li></a>
 +
                        </ul>
 +
                    </li>
 +
                    <li>
 +
                        <div class="ball"></div>
 +
                        <a href="" class="a1">Parts</a>
 +
                        <ul class="two">
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Parts"><li>Parts</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Contribution"><li>Contributions</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Engineering"><li>Engeering Success</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Improve"><li>Improve</li></a>
 +
                        </ul>
 +
 
 +
                    </li>
 +
                    <li>
 +
                        <div class="ball"></div>
 +
                        <a href="" class="a1">Human Practices</a>
 +
                        <ul class="two">
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Human_Practices"><li>Overview</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Human_Practices#Silver"><li>Silver</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Human_Practices#Gold"><li>Gold</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Education"><li>Science Communication</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Entrepreneurship"><li>Supporting Entrepreneurship</li></a>
 +
                        </ul>
 +
                    </li>
 +
                    <li>
 +
                        <div class="ball"></div>
 +
                        <a href="" class="a1">Awards</a>
 +
                        <ul class="two">
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Judging"><li>Judging</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Human_Practices"><li>Best Integrated Human Practices</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Education"><li>Best Education</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Entrepreneurship"><li>Best Supporting Entrepreneurship</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Model"><li>Best Model</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Hardware"><li>Best Hardware</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Entrepreneurship#Basic"><li>Best Basic Parts</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Entrepreneurship#Composite"><li>Best Composite Parts</li></a>
 +
                            <a href="https://2020.igem.org/Team:XMU-China/Entrepreneurship#Collection"><li>Best Part Collection</li></a>
 +
                        </ul>
 +
                    </li>
 +
                </ul>
 +
            </div>
 +
        </div>
 +
    </div>
 +
</div>
 +
<div class="main">
 +
    <section id="P0" class="js-scroll-step">
 +
        <div class="headline">
 +
            Kill Switch
 +
        </div>
 +
        <p style="font-weight: bold">1、The toxicity of MazF</p>
 +
        <p>The premise of our kill switch is to verify the toxicity of MazF. The toxicity of MazF cannot be reflected by OD600, viability count of colony-forming unit (CFU) was employed to investigate if MazF can kill <i>E. coli</i>.</p>
 +
        <p>pBAD/araC is chosen to express MazF. the plasmid carrying pBAD/araC-RBS-MazF-Terminator (BBa_K3332083) were transformed into E. coli BL21 (DE3) and 0.2% arabinose was added into the culture medium as an inducer in the induction group. Compared with that in the non-induction group (Figure 1 A1-4), MazF inhibited the growth of E. coli stronger from 6 hours to 12 hours in the induction group (Figure 1 B1-4). However, the phenomenon described above was not obvious 12 hours later.  </p>
 +
        <p class="F3"><img src="https://static.igem.org/mediawiki/2020/d/db/T--XMU-China--XMU-China_2020-CFU.png"></p>
 +
        <p class="Figure_word">Figure 1 The picture of CFU of pBAD/araC-RBS-MazF-Terminator (A is the non-induction group and B is the induction group)</p>
 +
        <p class="F3"><img src="https://static.igem.org/mediawiki/2020/b/b0/T--XMU-China--XMU-China_2020-RESULT.png"></p>
 +
        <p class="Figure_word">Figure 2 The results of CFU of pBAD/araC-RBS-MazF-Terminator</p>
 +
    </section>
 +
    <section id="P1" class="js-scroll-step">
 +
        <div class="headline">
 +
            2、The kill effect of pBAD/araC-Inverter-MazF
 +
        </div>
 +
        <p>In order to make sure that the engineered E. coli was killed in the environment without arabinose, Inverter was inserted into the circuit. Similarly, the viability count of CFU was chosen to verify the performance of our suicide switch. The plasmid carrying pBAD/araC-Inverter-RBS-MazF-Terminator (BBa_K3332081) were transformed into E. coli BL21 (DE3) and 0.2% arabinose was added into the culture medium as an inducer in the induction group. As shown in Figure 3, the growth of E. coli in the non-induction group (Figure 3 C1-4) was inhibited significantly after 6 hours. What’s more, there were no colonies left on the petri plate after 15 hours. That is to say, our kill switch has a good performance in killing E. coli when there is no arabinose. Also, we should pay attention to the reduction of the number of E. coli in the induction group (Figure 3 D1-4), which indicated the leakage of MazF. Although the leakage at a low level.</p>
 +
        <p class="F3"><img src="https://static.igem.org/mediawiki/2020/e/e8/T--XMU-China--XMU-China_2020-PICTURE.png"></p>
 +
        <p class="Figure_word">Figure 3 The picture of CFU of pBAD/araC-Inverter-RBS-MazF-Terminator (C is the non-induction group and D is the induction group)</p>
 +
        <p class="F3"><img src="https://static.igem.org/mediawiki/2020/4/48/T--XMU-China--XMU-China_2020-RESULT2.png"></p>
 +
        <p class="Figure_word">Figure 4 The result of CFU of pBAD/araC-Inverter-RBS-MazF-Terminator</p>
 +
    </section>
 +
</div>
 +
</body>
 +
<!-- <script type="text/javascript" src="js/right.js"></script> -->
 +
<script src="https://2020.igem.org/Team:XMU-China/js/nav1?action=raw&amp;ctype=text/javascript"></script>
 +
<script src="https://2020.igem.org/Team:XMU-China/js/nav2?action=raw&amp;ctype=text/javascript"></script>
 +
<div class="footer">
 +
    <div class="footer_bottom">
 +
        <p>Xiamen University, Fujian, China</p>
 +
        <p>No.422, Siming South Road, Fujian, P.R.China 361005</p>
 +
        <a href="#top"><img src="https://static.igem.org/mediawiki/2020/1/1a/T--XMU-China--returntop.png" class="fuix"></a>
 +
    </div>
 +
</div>
 +
</html>

Revision as of 23:43, 27 October 2020

Kill Switch

1、The toxicity of MazF

The premise of our kill switch is to verify the toxicity of MazF. The toxicity of MazF cannot be reflected by OD600, viability count of colony-forming unit (CFU) was employed to investigate if MazF can kill E. coli.

pBAD/araC is chosen to express MazF. the plasmid carrying pBAD/araC-RBS-MazF-Terminator (BBa_K3332083) were transformed into E. coli BL21 (DE3) and 0.2% arabinose was added into the culture medium as an inducer in the induction group. Compared with that in the non-induction group (Figure 1 A1-4), MazF inhibited the growth of E. coli stronger from 6 hours to 12 hours in the induction group (Figure 1 B1-4). However, the phenomenon described above was not obvious 12 hours later.

Figure 1 The picture of CFU of pBAD/araC-RBS-MazF-Terminator (A is the non-induction group and B is the induction group)

Figure 2 The results of CFU of pBAD/araC-RBS-MazF-Terminator

2、The kill effect of pBAD/araC-Inverter-MazF

In order to make sure that the engineered E. coli was killed in the environment without arabinose, Inverter was inserted into the circuit. Similarly, the viability count of CFU was chosen to verify the performance of our suicide switch. The plasmid carrying pBAD/araC-Inverter-RBS-MazF-Terminator (BBa_K3332081) were transformed into E. coli BL21 (DE3) and 0.2% arabinose was added into the culture medium as an inducer in the induction group. As shown in Figure 3, the growth of E. coli in the non-induction group (Figure 3 C1-4) was inhibited significantly after 6 hours. What’s more, there were no colonies left on the petri plate after 15 hours. That is to say, our kill switch has a good performance in killing E. coli when there is no arabinose. Also, we should pay attention to the reduction of the number of E. coli in the induction group (Figure 3 D1-4), which indicated the leakage of MazF. Although the leakage at a low level.

Figure 3 The picture of CFU of pBAD/araC-Inverter-RBS-MazF-Terminator (C is the non-induction group and D is the induction group)

Figure 4 The result of CFU of pBAD/araC-Inverter-RBS-MazF-Terminator

Xiamen University, Fujian, China

No.422, Siming South Road, Fujian, P.R.China 361005