Difference between revisions of "Team:UMaryland/Design"

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{{Team:UMarylandClasses}}
 
{{Team:UMarylandClasses}}
 
{{Team:UMarylandMenu}}
 
{{Team:UMarylandMenu}}
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<html>
 
<html>
  
 
<!--Attention! If you are not part of the website team, you are NOT allowed to touch anything above this line without the express permission of Best Kohai.-->
 
<!--Attention! If you are not part of the website team, you are NOT allowed to touch anything above this line without the express permission of Best Kohai.-->
  
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<head>
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<style type="text/css">
  
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<style>
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h1,h2,h3,h4,p,a {
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text-decoration: none;
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}
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h1 {
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font-size: 250%;
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}
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h2 {
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font-size: 200%;
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}
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h3 {
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font-size: 150%;
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}
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h4 {
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font-size: 125%;
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}
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p {
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font-size: 100%;
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}
  
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</style>
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#layer1 {
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overflow:hidden;
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position:relative;
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width:100%;
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margin:0px;
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padding:20px;
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background-image: -webkit-linear-gradient(top, #FFFFFF 0%, #41CC6B 50%, #FFFFFF 100%);
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min-height:42px;
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}
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#layer2 {
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overflow:hidden;
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position:relative;
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width:100%;
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margin:0px;
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padding:20px;
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background-image: -webkit-linear-gradient(top, #FFFFFF 0%, #46DBF2 50%, #FFFFFF 100%);
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min-height:42px;
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}
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#layer3 {
 +
overflow:hidden;
 +
position:relative;
 +
width:100%;
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margin:0px;
 +
padding:20px;
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background-image: -webkit-linear-gradient(top, #FFFFFF 0%, #E6AC39 50%, #FFFFFF 100%);
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min-height:42px;
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}
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#layer4 {
 +
overflow:hidden;
 +
position:relative;
 +
width:100%;
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margin:0px;
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padding:20px;
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background-image: -webkit-linear-gradient(top, #FFFFFF 0%, #FB7AFF 50%, #FFFFFF 100%);
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min-height:42px;
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}
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#contentbox{
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width: 80%;
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max-width: 1900px;
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min-width: 1000px;
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margin:auto;
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}
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#cover {
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overflow:hidden;
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position:relative;
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width:100%;
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margin:auto;
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padding:0px;
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background-image: url("http://i.imgur.com/YFUpQkT.jpg");
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background-size: 100% ;
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background-repeat: no-repeat;
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height: 100%;
 +
width:100%;
 +
min-height:1109;
 +
min-width:675;
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color: #0DFF00;
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font-family: Tahoma, Geneva, sans-serif;
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font-size:xx-large;
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text-align: center;
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bottom:0px;
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}
  
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#button {
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overflow:hidden;
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display: inline-block;
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height:120px;
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width: 240px;
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background-image: -webkit-linear-gradient(top, #0DFF00 0%, #FFFFFF 50%, #FFFF3B 100%);
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border: 2px solid black;
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border-radius: 10px;
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margin: 25px;
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padding:10px;
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font-size:medium;
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color:black;
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text-align: center;
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}
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</style>
  
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<link rel="stylesheet" type="text/css" href="https://2015.igem.org/Team:UMaryland/bootstrap.css?action=raw&ctype=text/css">
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<div id='cover'>
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<link rel="stylesheet" type="text/css" href="https://2015.igem.org/Team:UMaryland/UMD_description.css?action=raw&ctype=text/css">
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<br><br><br><br>
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</head>
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<br>
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<p style="font-size:72px"><u><b>Project Design: Hok/Sok</b></u></style>
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<body>
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<br>
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<br>
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<!-- HTML -->  
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<div id='swagcontent'>
 
  
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<a href = "#Hok">
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</div>
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<div class = "btn btn-primary btn-lg col-xs-10 col-xs-offset-1 col-sm-3 col-sm-offset-2">
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  <div class = "wrapper">Hok/Sok</div>
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</div>
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</a>  
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<div style = "background-color: #FFE7CE; padding:8px; border-radius: 7px ">
 
  
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<a href = "#Lutein">
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<div id='layer1'>
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<div class = "btn btn-primary btn-lg col-xs-10 col-xs-offset-1 col-sm-3 col-sm-offset-2">
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<div id='contentbox'>
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  <div class = "wrapper">Lutein</div>
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<h1><b>Hok/Sok Construct</b></h1>
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</div>
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</a>
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<div style="clear:both">
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−
 
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<a name = "Hok">  
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<h1> Design </h1>
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</a>
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<h2>Hok-Sok</h2>
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<h3>What is Hok-Sok?</h3>
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<h3>Hok-Sok Construct</h3>
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<ul>
 
<ul>
 
<li> 1. Full Hok-Sok sequence taken from Gerdes et al. Sequence is originally from the R-plasmid of <i>E. coli K-12</i>.</li>
 
<li> 1. Full Hok-Sok sequence taken from Gerdes et al. Sequence is originally from the R-plasmid of <i>E. coli K-12</i>.</li>
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</ul>
 
</ul>
  
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<h3>Fluorescence Studies</h3>
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</div>
 +
</div>
 +
 
 +
<div id='layer2'>
 +
<div id='contentbox'>
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<a name="HS"><h1><b>Fluorescence Studies</b></h1></a>  
 +
 
 
<ul>
 
<ul>
 
<li>1. Hok-Sok was coupled to a constitutive generator of unstable RFP to form a larger composite part.</li>
 
<li>1. Hok-Sok was coupled to a constitutive generator of unstable RFP to form a larger composite part.</li>
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</ul>
 
</ul>
  
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<h3>Plating Studies</h3>
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</div>
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</div>
 +
 
 +
<div id='layer3'>
 +
<div id='contentbox'>
 +
<a name="PCR"><h1>Plating Studies</h1></a>  
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<p>Along with measuring culture fluorescence, we also tested the ability of hok-sok to maintain a plasmid using daily chloramphenicol challenges. The protocol is as follows:</p>
 
<p>Along with measuring culture fluorescence, we also tested the ability of hok-sok to maintain a plasmid using daily chloramphenicol challenges. The protocol is as follows:</p>
 
<ol>
 
<ol>
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<p>For continuing generations of BL21 strain E. coli, we observed that on the plates for groups A and B, there was growth but no redness. If the bacteria were retaining the plasmids with the chloramphenicol resistance, the RFP gene should have been expressed and the colonies should fluoresce. We hypothesized that the chloramphenicol resistance gene was being recombined into the bacterial genome so the bacteria could therefore freely eject our inserted plasmids. As BL21 carries the gene for recombinase, it is possible. However, DH5α, as a common cloning strain, does not have recombinase. We created a new generation with every group (A, B, C, D, and E) to test whether the same plate would have similar results or once the bacteria stopped fluorescing, there would be no growth on the plates.</p>
 
<p>For continuing generations of BL21 strain E. coli, we observed that on the plates for groups A and B, there was growth but no redness. If the bacteria were retaining the plasmids with the chloramphenicol resistance, the RFP gene should have been expressed and the colonies should fluoresce. We hypothesized that the chloramphenicol resistance gene was being recombined into the bacterial genome so the bacteria could therefore freely eject our inserted plasmids. As BL21 carries the gene for recombinase, it is possible. However, DH5α, as a common cloning strain, does not have recombinase. We created a new generation with every group (A, B, C, D, and E) to test whether the same plate would have similar results or once the bacteria stopped fluorescing, there would be no growth on the plates.</p>
  
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<h3>Growth Curve</h3>
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</div>
 +
</div>
 +
 
 +
<div id='layer4'>
 +
<div id='contentbox'>
 +
<a name="PCR"><h1>Growth Curve</h1></a>  
 +
 
 
<p>We created a growth curve of Hok/Sok in comparison to controls to test the effectiveness of the Hok/Sok system in keeping the bacteria alive. We had four groups:
 
<p>We created a growth curve of Hok/Sok in comparison to controls to test the effectiveness of the Hok/Sok system in keeping the bacteria alive. We had four groups:
 
<ul>
 
<ul>
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<p>We started growing 250 mL cultures and monitored the OD at 600nm using a spectrophotometer over the span of 7.5 hours. </p>
 
<p>We started growing 250 mL cultures and monitored the OD at 600nm using a spectrophotometer over the span of 7.5 hours. </p>
  
 +
</div>
 +
</div>
 +
 +
<div id='layer1'>
 +
<div id='contentbox'>
 +
<a name="PCR"><h1>Parts Referenced</h1></a>
  
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<p><b>Parts Referenced</b></p>
 
 
<ul>
 
<ul>
 
<li><a href = http://parts.igem.org/Part:BBa_K1783001>Hok-Sok Type I Toxin-Antitoxin Cassette</a></li>
 
<li><a href = http://parts.igem.org/Part:BBa_K1783001>Hok-Sok Type I Toxin-Antitoxin Cassette</a></li>
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<br>
 
<br>
  
 +
</div>
  
 +
</div>
 
</div>  
 
</div>  
  

Revision as of 17:02, 11 September 2015