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| | <style> | | <style> |
| − | #top_title {display: none;} | + | #top_title {display: none;} |
| − | #sideMenu {display: none;} | + | #sideMenu {display: none;} |
| − | #content {margin: auto; padding: 0; width: 60%;} | + | #content {margin: auto; padding: 0; width: 100%; background:#3aafa9;} |
| | </style> | | </style> |
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| − | <center>
| + | </html> |
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| − | <head>
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| − | <title>East Chapel Hill Highschool iGem</title>
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| − | <meta charset="utf-8">
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| − | <meta name="viewport" content="width=device-width, initial-scale=1">
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| − | <link rel="stylesheet" href="https://maxcdn.bootstrapcdn.com/bootstrap/3.3.7/css/bootstrap.min.css">
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| − | <script src="https://ajax.googleapis.com/ajax/libs/jquery/3.2.1/jquery.min.js"></script>
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| − | <script src="https://maxcdn.bootstrapcdn.com/bootstrap/3.3.7/js/bootstrap.min.js"></script>
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| − | <style>
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| − | /* Remove the navbar's default margin-bottom and rounded borders */
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| − |
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| − | .navbar {
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| − | margin-bottom: 0;
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| − | margin-right: 0;
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| − | border-radius: 0;
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| − | width: 100%;
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| − | /*background-color: #ffcccc;*/
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| − | }
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| − | body {
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| − | background-color: black;
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| − | text-align: left;
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| − | font size: "12";
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| − | }
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| − | */
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| − | /* Set height of the grid so .sidenav can be 100% (adjust as needed) */
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| − | .row.content {height: 450px}
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| − | /* Set gray background color and 100% height */
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| − | .sidenav {
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| − | padding-top: 20px;
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| − | background-color: black;
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| − | height: 100%;
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| − | }
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| − | ul.yourclass
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| − | { display: inline-block;
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| − | padding-rigth: 15px;
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| − | }
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| − | /* Set black background color, white text and some padding */
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| − | footer {
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| − | background-color: #555;
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| − | color: white;
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| − | padding: 15px;
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| − | }
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| − | /* On small screens, set height to 'auto' for sidenav and grid */
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| − | @media screen and (max-width: 767px) {
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| − | .sidenav {
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| − | height: auto;
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| − | padding: 15px;
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| − | }
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| − | .row.content {height:auto;}
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| − | }
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| − | </style>
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| − | </head>
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| − | <body>
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| − | <nav class="navbar navbar-inverse">
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| − | <div class="container-fluid">
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| − | <div class="navbar-header">
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| − | <button type="button" class="navbar-toggle" data-toggle="collapse" data-target="#myNavbar">
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| − | <span class="icon-bar"></span>
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| − | <span class="icon-bar"></span>
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| − | <span class="icon-bar"></span>
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| − | </button>
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| − | </div>
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| − | <div class="collapse navbar-collapse" id="myNavbar">
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| − | <ul class="nav navbar-nav">
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| − | <li><img align="middle" style="width:100%" height="50px" src="https://static.igem.org/mediawiki/2017/8/89/T--East_Chapel_Hill--Icon0.png"></img> </li>
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| − | </ul>
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| − | <ul class="nav navbar-nav navbar-right">
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| − | <li class="active"><a href="https://2017.igem.org/Team:East_Chapel_Hill">Home</a></li>
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| − | <li class="dropdown">
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| − | <a class="dropdown-toggle" data-toggle="dropdown" href="Team.html">Team </a>
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| − | <span class="caret"></span></a>
| + | |
| − | <ul class="dropdown-menu">
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| − | <li><a href="#">Team-part-1</a></li>
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| − | <li><a href="#">Team-part-2</a></li>
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| − | <li><a href="#">Team-part-3</a></li>
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| − | </ul>
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| − | </li>
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| − | <li><a href="Project.html">Project</a></li>
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| − | <li><a href="Results.html">Results</a></li>
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| − | <li><a href="Notebook.html">Notebook</a></li>
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| − | <li><a <href="https://2017.igem.org/Team:East_Chapel_Hill/testHumanPractices">Human Practices </a></li>
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| − | <li><a <href="https://2017.igem.org/Team:East_Chapel_Hill/Attributions">Sponsors</a></li>
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| − | </ul>
| + | |
| − | </div>
| + | |
| − | </nav>
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| − | <img style="width:100%" height="350px" src= "https://static.igem.org/mediawiki/2017/7/71/EastChapelHillWildType.png"> </img>
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| − | <p>Wild type and ∆crcB were grown directly next to each other at varying levels of fluoride. As described in literature, ∆crcB should not grow when exposed to fluoride levels over 500µM. Unexpectedly, ∆crcB did grow on high levels of fluoride when placed on the same plates as wild type E. coli. Without wild type E. coli, no growth of ∆crcB was seen on 1mM and 2mM fluoride. </p>
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| − | <p>In order to figure out the cause of the unexpected growth, we reached out to Dr. Randy Stockbridge at University of Michigan for possible explanations.
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| − | Dr. Stockbridge informed us of Fluoride’s dependability of pH. A low pH environment is necessary for Fluoride to enter cells. Therefore, we hypothesized that the wild type altered the pH of the agar to a more basic level, facilitating the growth of ∆crcB.</p>
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| − | | + | |
| − | <p>To confirm our hypothesis, we prepared similar set of plates, but with phenol red, a pH indicator, for visual confirmation. </p>
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| − | <img style="width:100%" height="350px" src= "https://static.igem.org/mediawiki/2017/d/d5/EastChapelHillRed.png"> </img>
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| − | <p>Phenol red appears red at basic pH and turns yellow at acidic pH. In the absence of a buffer, wild type has a visibly basic pH. Therefore, once the entire plate became basic, ∆crcB showed fair amount of growth. On previous days, ∆crcB grew only at red areas (low pH).
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| − | </p>
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| − | | + | |
| − | <p>To further confirm our results, we prepared the same plates with a MES buffer, which kept the agar at a 6.5 pH, which is slightly acidic.
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| − | </p>
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| − | <img style="width:100%" height="350px" src= "https://static.igem.org/mediawiki/2017/8/80/EastChapelHillMES.png"> </img>
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| − | <p>Upon the addition of the buffer, the alteration of pH was significantly delayed, so F was able to enter the membrane of ∆crcB, allowing the strain to grow.
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| − | </p>
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| − | | + | |
| − | <p>
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| − | CHOP (Chloramphenicol Operon) and ∆crcB (E. coli without Fluoride Channel) were added to 50µ chloramphenicol plates and observed over the course of 4 days.
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| − | | + | |
| − | <ul>
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| − | <li>The plates had increasing concentrations of F with CHOP and ∆crcB to determine the level of fluoride that leads to the greatest growth of CHOP. </li>
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| − | <li>∆crcB was part of the control and did not grow because it does not have the fluoride channel to pump out the F.</li>
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| − | <li>As observed, there is most growth of CHOP on 100μM of F, suggesting maximum growth of CHOP at 100μM of F.</li>
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| − | <li>Another set of plates with 10-5 and 10-4 dilutions of Chloramphenicol were prepared. </li>
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| − | <li>The plates were divided into quadrants with CHOP on the top left and bottom left quadrants and crcB on the top right and bottom right. </li>
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| − | <li>CHOP had the most growth at 100μM F at 10-4 dilution. </li>
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| − | </ul>
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| − | | + | |
| − | </p>
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| − | <h1> Future Directions
| + | |
| − | </h1>
| + | |
| − | <p>
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| − | The CHOP System can be used to improve the affinity and characterization of the fluoride riboswitch.
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| − | </p>
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| − | <p>Green Fluorescent Protein (GFP) system may be used for detecting presence of fluoride; at a certain level of fluoride, it can trigger gene expression of the GFP as the fluoride riboswitch releases terminator. This may provide a better visual mechanism for people to determine if their water can be safe to drink or not. </p>
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| − | <p>A fluorinase gene can be utilized to bio-remediate the excess fluoride in water. </p>
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| − |
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| − | <map name="homepage">
| + | |
| − | <area shape="circle" coords=450,450,150" href="Project.html" >
| + | |
| − | <area shape="circle" coords="750,150,150" href="Results.html">
| + | |
| − | </map>
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| − | | + | |
| − | | + | |
| − | </div>
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