PennState/Project/Dosimeter/InitialTest

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'''Promoter Test Constructs'''
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'''Dosimeter Test Construct'''
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To test each of the four xyl promoters, each is placed in a construct followed by a ribosome binding site, the reporter green florescent protein, and a terminator. The strength of each is measured by gfp emission.
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To test the dosimeter, the assembled lambda region is placed in a plasmid preceding a ribosome binding site, the reporter green florescent protein, and a terminator. The promoter's sensitivity to radiation can be readily tested with flow cell cytometry and fluorescence plate reading
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<html><img src="http://openwetware.org/images/5/54/Dosimeter_Test_Construct.jpg" id="logo" name="logo" height="128" width="800" alt="" /></html>This construct uses the xylF and xylA facing promoters that includes the CRP-cAMP binding region.
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<html><img src="http://openwetware.org/images/5/54/Dosimeter_Test_Construct.jpg" id="logo" name="logo" height="128" width="800" alt="" /><br></html>
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This construct uses the lambda operator to express GFP when induced by radiation.
 +
 
 +
However, to establish the "off" state which corresponds to the lysogenic state of the WT lambda phage, it is necessary to use a second plasmid that produces LVA tagged Cl upon IPTG induction. Additionally, the plasmid harboring this sequence is IPTG inducible.
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 +
 
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<html><img src="http://openwetware.org/images/d/d0/PSU_GFL_IGEM_Establishing_Plasmid.jpg" id="logo" name="logo" height="128" width="332" alt="" /><br></html>
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This construct uses the pLAC operator to produce LVA tagged Cl upon IPTG induction.
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<html>
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<!-- START OF LUC'S CODE -->
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<!-- START CSS: the following css needs to be on the page to style the podcast divs properly. Diagonal span background based on www.mezzoblue.com/zengarden/alldesigns/ -->
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<style type="text/css">
 +
.column {
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width:  200px;
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position: relative;
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display: block;
 +
float: left;
 +
clear: none;
 +
margin-right: 10px;
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}
 +
.podcast{
 +
position: relative;
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float: none;
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border: 1px dotted #c7cbce;
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height: 110px;
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background-color: #f1f1f1;
 +
clear: both;
 +
margin-bottom: 42px;
 +
font: 12px/1.4em "Lucida Grande", "Lucida Sans Unicode", Verdana, Tahoma, Arial, sans-serif;
 +
padding: 0;
 +
width: 175px;
 +
}
 +
.podcast h3{
 +
color: #3d3d3d;
 +
margin: .2em 0 .5em;
 +
padding: 0 0 0 4px;
 +
font-family: Verdana, sans-serif;
 +
font-size: 100%;
 +
}
 +
.podcast a:link, .podcast a:visited{
 +
text-decoration: none;
 +
color: #3d3d3d;
 +
}
 +
.podcast a:hover {
 +
color: #aaaaff;
 +
}
 +
a:hover.screenshot {
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}
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.podcast .screenshot {
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float: left;
 +
position: relative;
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clear: none;
 +
display: block;
 +
margin: 0px 0px 0px 16px;
 +
}
 +
.podcast .screenshot img {
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background: #ffffff;
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border: 1px solid #c7cbce;
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padding: 4px;
 +
}
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.podcast .screenshot span {
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position: absolute;
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bottom:0px;
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right: 1px;
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padding: 4px 20px 4px 26px;
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background: url(https://static.igem.org/mediawiki/2007/0/09/Podcast-download-bg.gif) no-repeat;
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font-size: 0.8em;
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line-height: 1;
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}
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.podcast .screenshot span a{
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padding: 0 0 0 0.5em;
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}
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.podcast .description {
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display: block;
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position: relative;
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float: left;
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width: 220px;
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margin-left: 3px;
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padding-left: 1.2em;
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}
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.podcast .description a{
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border-bottom: 1px dotted #aaaaff;
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}
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.podcast .description a:hover{
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border-bottom: 1px dotted #aaaaff;
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}
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.screenshot img:hover {
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border: 1px solid #aaaaff;
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}
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</style></html>
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<html>
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<!-- PODCAST HTML: the following html structures the information for each podcast. Each podcast entry is contained in its own podcast div, which are arranged into two columns by two column divs.  The last column must be cleared.-->
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<div class="column">
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<div class="podcast">
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<!-- this is where you need to post the link to where it will go (the second one)-->
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<h3><a rel="enclosure" href="https://2007.igem.org/PennState/Project/Dosimeter" ="window.popup_player_258600 = window.open('https://2007.igem.org/PennState/Project/Dosimeter','post_258600','toolbar=no,scrollbars=no,directories=no,resizable=yes,width=360,height=305,top=20,left=20,location=no,menubar=no,status=yes,'); return false;">
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Introduction
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</a></h3>
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<div class="screenshot">
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<!-- little white box thing --> <a rel="enclosure" href="https://2007.igem.org/PennState/Project/Dosimeter"
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<img src="http://openwetware.org/images/9/97/Psuigem2007promoterimage.jpg" width="133" height="100" alt="" />
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</a>
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<span class="download">
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<a href="http://openwetware.org/images/9/97/Psuigem2007promoterimage.jpg">Image Link</a>
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</span>
 +
</div>
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</div>
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 +
</div><div class="column">
 +
 
 +
<div class="podcast">
 +
<!-- this is where you need to post the link to where it will go (the second one)-->
 +
<h3><a rel="enclosure" href="https://2007.igem.org/PennState/Project/Dosimeter/InitialTest" ="window.popup_player_258600 = window.open('https://2007.igem.org/PennState/Project/Dosimeter/InitialTest','post_258600','toolbar=no,scrollbars=no,directories=no,resizable=yes,width=360,height=305,top=20,left=20,location=no,menubar=no,status=yes,'); return false;">
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Test Constructs
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</a></h3>
 +
<div class="screenshot">
 +
<!-- little white box thing --> <a rel="enclosure" href="https://2007.igem.org/PennState/Project/Dosimeter/InitialTest"
 +
<img src="http://openwetware.org/images/b/be/Psuigem2007RBSimage.jpg" width="133" height="100" alt="" />
 +
</a>
 +
<span class="download">
 +
<a href="http://openwetware.org/images/b/be/Psuigem2007RBSimage.jpg">Image Link</a>
 +
</span>
 +
</div>
 +
</div>
 +
</div>
 +
<div class="column"><div class="podcast">
 +
<!-- this is where you need to post the link to where it will go (the second one)-->
 +
<h3><a rel="enclosure" href="https://2007.igem.org/PennState/Project/Dosimeter/Dosimetry" ="window.popup_player_258600 = window.open('http://openwetware.org/images/c/c9/XylRdimer1.png','post_258600','toolbar=no,scrollbars=no,directories=no,resizable=yes,width=360,height=305,top=20,left=20,location=no,menubar=no,status=yes,'); return false;">
 +
Dosimetry
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</a></h3>
 +
<div class="screenshot">
 +
<!-- little white box thing --> <a rel="enclosure" href="https://2007.igem.org/PennState/Project/Dosimeter/Dosimetry"
 +
<img src="http://openwetware.org/images/2/21/Psuigem2007geneimage.jpg" width="133" height="100" alt="" />
 +
</a>
 +
<span class="download">
 +
<a href="http://openwetware.org/images/2/21/Psuigem2007geneimage.jpg">Image Link</a>
 +
</span>
 +
</div>
 +
</div>
 +
</div>
 +
<div class="column"><div class="podcast">
 +
<!-- this is where you need to post the link to where it will go (the second one)-->
 +
<h3><a rel="enclosure" href="https://2007.igem.org/PennState/Project/Dosimeter/FinalConstructs" ="window.popup_player_258600 = window.open('https://2007.igem.org/PennState/Project/Dosimeter/FinalConstructs','post_258600','toolbar=no,scrollbars=no,directories=no,resizable=yes,width=360,height=305,top=20,left=20,location=no,menubar=no,status=yes,'); return false;">
 +
Final Constructs
 +
</a></h3>
 +
<div class="screenshot">
 +
<!-- little white box thing --> <a rel="enclosure" href="https://2007.igem.org/PennState/Project/Dosimeter/FinalConstructs"
 +
<img src="http://openwetware.org/images/9/9b/Psuigem2007TTimage.jpg" width="133" height="100" alt="" />
 +
</a>
 +
<span class="download">
 +
<a href="http://openwetware.org/images/9/9b/Psuigem2007TTimage.jpg">Image Link</a>
 +
</span>
 +
</div>
 +
</div>
 +
 
 +
<div style="clear:both"></div>
 +
<!-- END OF LUC'S CODE -->
 +
</html>

Latest revision as of 03:20, 27 October 2007

Dosimeter Test Construct

To test the dosimeter, the assembled lambda region is placed in a plasmid preceding a ribosome binding site, the reporter green florescent protein, and a terminator. The promoter's sensitivity to radiation can be readily tested with flow cell cytometry and fluorescence plate reading



This construct uses the lambda operator to express GFP when induced by radiation.

However, to establish the "off" state which corresponds to the lysogenic state of the WT lambda phage, it is necessary to use a second plasmid that produces LVA tagged Cl upon IPTG induction. Additionally, the plasmid harboring this sequence is IPTG inducible.



This construct uses the pLAC operator to produce LVA tagged Cl upon IPTG induction.

Introduction

Image Link

Test Constructs

Image Link

Dosimetry

Image Link

Final Constructs

Image Link

Retrieved from "http://2007.igem.org/wiki/index.php/PennState/Project/Dosimeter/InitialTest"