Team:Penn State/Bidirectional Promoters Results

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     <h2>Bidirectional Promoters</h2>
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     <h3>Bidirectional Promoters</h3></br>
     <p>Scientists are frequently confounded by wayward promoters; that is, promoters which do not produce  
     <p>Scientists are frequently confounded by wayward promoters; that is, promoters which do not produce  
the expected proteins. Some bidirectional promoters are known to exist, but which way they promote  
the expected proteins. Some bidirectional promoters are known to exist, but which way they promote  
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<h3>Data Collection</h3>
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<h3>Palindromic Promoter Circuits</h3>
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    <p>Several BioBrick promoters have been tested in the bidirectional promoter construct. All successful ligations were cultured and the fluorescence of each individual promoter was tested in a TECAN machine.</P>
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<p><img src="https://static.igem.org/mediawiki/2012/f/fc/BiDirresults.png"></p>
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<p>The following promoters have been successfully tested for quantitative directionality using the TECAN. For a <a href="https://2012.igem.org/Team:Penn_State/Parts">complete list</a> of promoters we intend to test, please visit our <a href="https://2012.igem.org/Team:Penn_State/Parts">Parts</a> webpage.</p>
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<p>The graph above represents the relative fluorescence measured in a TECANM1000 instrument of several constructs containing various promoter inserts. The reporters indicate the directionality of each promoter sequence measured in the construct. The single biobrick ligated into the construct shows almost complete preference for the forward direction, while three palindromic sequences designed and ligated in the construct demonstrate varying amounts of bidirectionality within the DH10B E.coli cell. Bidirectional promoters are possible within the cell, and those designing promoters should be beware of palindromic sequences in their design and their potential effects on the initiation of protein within the cell. </p>
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<p>Data collection and results are demonstrated graphically below for J23102, and three custom palindrome promoters. (Click on the graphs to see a larger version of the graph.)</p>
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<h3>Further Application</h3>
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GFP Expression of all samples: <br/>
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<p>Further characterization of both biobrick promoters and palindromic sequences will yield more information of the prevalence of bidirectionality in promoters. To view the sequences we plan to test in this construct, please visit our <a href="https://2012.igem.org/Team:Penn_State/Parts">parts</a> page.</p>
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<a href="https://static.igem.org/mediawiki/2012/3/3c/PennState_BiDir_GFP_All_Graph.jpg">
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Average GFP Expression of all samples: <br/>
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<a href="https://static.igem.org/mediawiki/2012/a/a9/PennState_BiDir_GFP_Combined_Graph.jpg">
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<img src="https://static.igem.org/mediawiki/2012/a/a9/PennState_BiDir_GFP_Combined_Graph.jpg" width="300px" height="180px">
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RFP Expression of all samples: <br/>
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<a href="https://static.igem.org/mediawiki/2012/b/b6/PennState_BiDir_RFP_All_Graph.jpg">
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Average RFP Expression of all samples: <br/>
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<a href="https://static.igem.org/mediawiki/2012/9/9f/PennState_BiDir_RFP_Combined_Graph.jpg">
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     <p><strong>&copy; Hannah Jepsen-Burger<a href="#"></a></strong> | Follow us on Twitter<a href="#"></a></p>
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     <p><strong>&copy; Hannah Jepsen-Burger</strong> | Follow us on Twitter<a href="#"></a></p>
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Latest revision as of 03:53, 27 October 2012

Bidirectional Promoters Overview

Bidirectional Promoters


Scientists are frequently confounded by wayward promoters; that is, promoters which do not produce the expected proteins. Some bidirectional promoters are known to exist, but which way they promote and the degree of expression has not been quantified. This project will test the directionality of several BioBrick promoters to answer these questions.

Bidirectional Promoters

Sample navigation menu:

Overview | Design | Results

Palindromic Promoter Circuits

The graph above represents the relative fluorescence measured in a TECANM1000 instrument of several constructs containing various promoter inserts. The reporters indicate the directionality of each promoter sequence measured in the construct. The single biobrick ligated into the construct shows almost complete preference for the forward direction, while three palindromic sequences designed and ligated in the construct demonstrate varying amounts of bidirectionality within the DH10B E.coli cell. Bidirectional promoters are possible within the cell, and those designing promoters should be beware of palindromic sequences in their design and their potential effects on the initiation of protein within the cell.

Further Application

Further characterization of both biobrick promoters and palindromic sequences will yield more information of the prevalence of bidirectionality in promoters. To view the sequences we plan to test in this construct, please visit our parts page.