Difference between revisions of "Team:OUC-China/Improve"

 
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{{OUC-China}}
 
{{OUC-China}}
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{{OUC-China/CSS}}
 
<html>
 
<html>
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<head>
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<meta charset="utf-8">
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<title>Improve</title>
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<style type="text/css">
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.ouc-right{position: relative;}
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.ouc-rightnav{position:absolute;right: -200%;top: 0; display: none;background-color: white;width: 200%; z-index: 2001}
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.ouc-right:hover .ouc-rightnav{display: inherit;}
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.ouc-rightnav a{background-color: white; color: #008EA1; width: 100%;height: 40px; line-height: 40px;}
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.ouc-rightnav a:visited {color: #008EA1;text-decoration: none; }
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.ouc-rightnav a:hover {background-color:#D2D2D2; color: white;text-decoration: none; }
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.ouc-vicedown{position: absolute; top:50px; }
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.ouc-vicedown a{display: block;height: 40px; width: 100%;text-decoration: none;color: #008EA1; line-height: 40px;text-align: center;}
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.ouc-vicedown a:visited{color:#008EA1;text-decoration: none }
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.ouc-vicedown a:hover{color:  #EDEC8Cl}
  
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.ouc-nav{color:white; background-color: #008EA1;display:block; width: 100%; color: white;padding: 0}
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.ouc-nav:visited{text-decoration: none; color: white}
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.ouc-nav:hover{text-decoration: none; background-color: white; color:#008EA1; }
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.ouc-navbar{position: relative;padding: 0;margin: 0;}
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.ouc-navbar:hover .ouc-down{display: inherit;}
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.ouc-down{position: absolute; top:50px;display: none;background-color: white; z-index: 1001}
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.ouc-navdown{color: #008EA1;display: block; width: 100%;height: 40px;}
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.ouc-navdown:visited{text-decoration: none;color:#008EA1; }
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.ouc-navdown:hover{background-color:#D2D2D2;color: white;text-decoration: none; z-index: 1501}
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.ouc-guide{position: fixed; width: 100%;top: 15px;z-index:1000}
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.ouc-page-header{border-bottom-color: #EDEC8C;border-bottom-width: thick;color: #008F75;}
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.top{position: fixed;bottom:0;right: 0;}
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.top:hover{animation: upp 0.6s ;}
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@keyframes upp {
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0% {margin-bottom: 0;}
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50% {margin-bottom: 80px;}
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100% {margin-bottom: 0;}
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}
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.ouc-reserve{background-color: #008EA1;color: white;}
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    .ouc-heading {margin: 0;color: #008EA1;}
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</style>
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</head>
  
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<body>
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<!--此处是导航-->
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<div class="ouc-guide" style="height: 50px;background-color:#008EA1 ;">
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<div class="row">
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<div class="col-md-2 navbar-header" style="text-align: center; padding: 0">
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<button class="navbar-toggle" type="button" data-toggle="collapse" data-target=".navbar-responsive-collapse">
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        <span class="sr-only">Toggle Navigation</span>
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        <span class="caret" style="color: white"></span>
  
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<a href="https://2017.igem.org/Team:OUC-China"><img src="https://static.igem.org/mediawiki/2017/8/89/T--OUC-China--logo.jpg" height="50px"/></a></div>
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<div class="collapse navbar-collapse navbar-responsive-collapse" style="padding: 0">
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<div class="col-md-1" style="padding: 0"></div>
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<div class="col-md-1 ouc-navbar" style="padding: 0">
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    <a href="##" style="line-height: 50px;text-align: center" class="ouc-nav">Team<span class="caret"></span></a>
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    <div class="ouc-down" style=" width: 100%; text-align: center">
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    <div><a href="https://2017.igem.org/Team:OUC-China/Team" style="line-height: 40px; " class="ouc-navdown">Members</a></div>
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    <div><a href="https://2017.igem.org/Team:OUC-China/Collaborations" style="line-height: 40px; " class="ouc-navdown">Collaborations</a></div>
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    <div><a href="https://2017.igem.org/Team:OUC-China/Attributions" style="line-height: 40px; " class="ouc-navdown">Attributions</a></div>
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    </div>
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</div>
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<div class="col-md-1 ouc-navbar" style="padding: 0">
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    <a href="##" style="line-height: 50px;text-align: center" class="ouc-nav">Project<span class="caret"></span></a>
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    <div class="ouc-down" style=" width: 100%; text-align: center">
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    <div><a href="https://2017.igem.org/Team:OUC-China/Description" style="line-height: 40px; " class="ouc-navdown">Description</a></div>
 +
    <div><a href="https://2017.igem.org/Team:OUC-China/Design" style="line-height: 40px; " class="ouc-navdown">Design</a></div>
 +
                <div class="ouc-right">
 +
        <a style="line-height: 40px; font-size: 15px;" class="ouc-navdown" href="https://2017.igem.org/Team:OUC-China/proof1">Proof of concept</a>
 +
        <div class="ouc-rightnav">
 +
        <a href="https://2017.igem.org/Team:OUC-China/proof1" style="display: block">Basic fermentation</a>
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        <a href="https://2017.igem.org/Team:OUC-China/proof2" style="display: block">Adhesion platform</a>
 +
        <a href="https://2017.igem.org/Team:OUC-China/proof3" style="display: block">MINI-GRE</a>
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        </div>
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    </div>
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    <div><a href="https://2017.igem.org/Team:OUC-China/Demonstrate" style="line-height: 40px; " class="ouc-navdown">Demonstration</a></div>
 +
    <div><a href="https://2017.igem.org/Team:OUC-China/InterLab" style="line-height: 40px; " class="ouc-navdown">InterLab</a></div>
 +
    <div><a href="https://2017.igem.org/Team:OUC-China/Improve" style="line-height: 40px; " class="ouc-navdown">Improvement</a></div>
 +
    <div><a href="https://2017.igem.org/Team:OUC-China/Notebook" style="line-height: 40px; " class="ouc-navdown">Notebook</a></div>
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    </div>
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</div>
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<div class="col-md-1" style="padding: 0"><a href="https://2017.igem.org/Team:OUC-China/Model" style="line-height: 50px;text-align: center" class="ouc-nav">Model</a></div>
 +
<div class="col-md-1" style="padding: 0"><a href="https://2017.igem.org/Team:OUC-China/Parts" style="line-height: 50px;text-align: center" class="ouc-nav">Parts</a></div>
 +
<div class="col-md-1" style="padding: 0"><a href="https://2017.igem.org/Team:OUC-China/Safety" style="line-height: 50px;text-align: center" class="ouc-nav">Safety</a></div>
 +
<div class="col-md-2 ouc-navbar" style="padding: 0">
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    <a href="https://2017.igem.org/Team:OUC-China/Engagement" style="line-height: 50px;text-align: center" class="ouc-nav">Human Practice<span class="caret"></span></a>
 +
    <div class="ouc-down" style=" width: 100%; text-align: center">
 +
    <div><a href="https://2017.igem.org/Team:OUC-China/HumanPractice" style="line-height: 40px; " class="ouc-navdown">Overview</a></div>
 +
    <div><a href="https://2017.igem.org/Team:OUC-China/HP/Silver" style="line-height: 40px; " class="ouc-navdown">Silver</a></div>
 +
    <div><a href="https://2017.igem.org/Team:OUC-China/HP/Gold_Integrated" style="line-height: 40px; " class="ouc-navdown">Gold and integrated</a></div>
 +
    <div><a href="https://2017.igem.org/Team:OUC-China/Engagement" style="line-height: 40px; " class="ouc-navdown">Engagement</a></div>
 +
    </div>
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</div>
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<div class="col-md-2" style="padding: 0"></div>
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</div>
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</div>
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</div>
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<!--此处是导航-->
  
−
<div class="column full_size">
 
−
<h1>Improve</h1>
 
−
<p>For teams seeking to improve upon a previous part or project, you should document all of your work on this page. Please remember to include all part measurement and characterization data on the part page on the Regisrty. Please include a link to your improved part on this page.</p>
 
  
−
<h3>Gold Medal Criterion #2</h3>
+
<div class="container">
−
<p><b>Standard Tracks:</b> Improve the function of an existing BioBrick Part. The original part must NOT be from your 2017 part number range. If you change the original part sequence, you must submit a new part. In addition, both the new and original part pages must reference each other. This working part must be different from the part documented in bronze #4 and silver #1.
+
    <div class="page-header ouc-page-header">
 +
        <h1><strong>Improvement</strong></h1>
 +
    </div>
  
−
<br><br>
+
<p style="font-size: 20px">
−
<b>Special Tracks:</b> Improve the function of an existing iGEM project (that your current team did not originally create) and display your achievement on your wiki.</p>
+
      This year, our team improved the promoter <a href = "http://parts.igem.org/Part:BBa_J23108" style = “color:blue;">BBa_J23018</a> in a novel way. The newly constructed promoter is <a href = "http://parts.igem.org/Part:BBa_K2314212" style = “color:blue;">BBa_K2314212</a>, in which we add a UTRrpsT sequence and enhanced its strength. Considering relative researches and the result of our experiments, we have the reason to believe that UTRrpsT has the potential to enhance the promoter as a universal enhancement module.
  
 +
    <h2 class="ouc-heading"><strong>Inspiration</strong></h2>
 +
    <div class="row">
 +
    <div class="col-md-4" style="text-align: center;">
 +
    <img src="https://static.igem.org/mediawiki/2017/f/fb/T--OUC-China--improve.5%E2%80%98UTR.png" width="400px"/>
 +
   
 +
    </div>
 +
    <div class="col-md-8">
 +
    <p style="font-size: 20px">
 +
    <br/><br/><br/><br/>
 +
    As known to all, 5'UTR will be transcribed and can be used as a regulatory element to adjust
 +
the translation process. Therefore, it is believed that a delicate
 +
structure of the 5'UTR has the potential as a universal enhancement
 +
module. We see that Zhou et al. screened some 5'UTR sequences from <i>E.
 +
coli</i> and helped improve the expression of target protein. Thinking about our need to express target gene in our project, we decided to edit an existing promoter in Part in this way. Hopefully, the strength will be enhanced. Since the promoter we picked was not operated this way before, we were not sure about its result at first, so we consulted the auther of our basic paper. Luckily,the idea also received the
 +
author's support.[1]
 +
    </p>
 +
    </div>
 +
    </div>
 +
   
 +
    <h2 class="ouc-heading"><strong>Design</strong></h2>
 +
    <p style="text-align: center">
 +
    <img src="https://static.igem.org/mediawiki/2017/thumb/a/aa/T--OUC-China--improve.illustration.png/800px-T--OUC-China--improve.illustration.png" width="700px"/>
 +
    </p>
 +
    <p style="font-size: 20px">
 +
    We selected BBa_J23108 from constitutive
 +
promoter family  and added a
 +
5'UTR sequence that we choose from the paper mentioned above thereafter. Fluorescent
 +
protein is used to characterize whether or not the
 +
5'UTR has an enhanced effect on the
 +
promoter.
 +
    </p>
 +
 
 +
 
  
 +
 +
    <h2 class="ouc-heading"><strong>Proof of concept</strong></h2>
 +
    <p style="text-align: center">
 +
    <img src="https://static.igem.org/mediawiki/2017/thumb/7/77/T--OUC-China--improve.FI_Abs600.png/800px-T--OUC-China--improve.FI_Abs600.png" width="700px"/>
 +
    </p>
 +
    <p style="font-size: 20px">
 +
    We constructed a circuit devices with a commonly used red fluorescent protein and  randomly
 +
selected the promoter and the terminator. The difference is that UTRrpsT is added to a loop and
 +
the fluorescence intensity is measured using a microplate reader and plotted. We found that the
 +
fluorescence intensity at 6h increased to 1.5 fold. This indicates that UTRrpsT has the capability
 +
to function as a generic enhancement module.（Error bars represent standard deviation of three biological replicates.）
 +
<h2 class="ouc-heading"><strong>Future work</strong></h2>
 +
        <p style="font-size: 20px">
 +
        According to the analysis of data, we may draw a conclusion that the part can impove the expression of our target protein (mRFP). In the future, we can change different reporters to  detect whether different  reporters may have the same result---that their expression will all be improve. If so, we can make sure that the part is universal for improving all proteins and we can utilize this provement to increase the expression of mSA, which improve the probability of binding to yeast successfully and strengthen the binding. Further, we might offer help to other teams.
 +
    </p>
 +
    <h2 class="ouc-heading"><strong>Reference</strong></h2>
 +
[1]Zhou, Shenghu, et al. "Obtaining a Panel of Cascade Promoter-5′-UTR Complexesin Escherichia coli." Acs Synthetic Biology 6.6(2017)<br/>
 
</div>
 
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<a href="#top"><img class="top" src="https://static.igem.org/mediawiki/2017/b/b7/T--OUC-China--top.jpeg" width="101" height="101" /></a>
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 +
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 +
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Latest revision as of 13:58, 18 November 2017

Improve
Team
Project
Proof of concept
Basic fermentation Adhesion platform MINI-GRE
Model
Parts
Safety
Human Practice

Improvement

This year, our team improved the promoter BBa_J23018 in a novel way. The newly constructed promoter is BBa_K2314212, in which we add a UTRrpsT sequence and enhanced its strength. Considering relative researches and the result of our experiments, we have the reason to believe that UTRrpsT has the potential to enhance the promoter as a universal enhancement module.

Inspiration





As known to all, 5'UTR will be transcribed and can be used as a regulatory element to adjust the translation process. Therefore, it is believed that a delicate structure of the 5'UTR has the potential as a universal enhancement module. We see that Zhou et al. screened some 5'UTR sequences from E. coli and helped improve the expression of target protein. Thinking about our need to express target gene in our project, we decided to edit an existing promoter in Part in this way. Hopefully, the strength will be enhanced. Since the promoter we picked was not operated this way before, we were not sure about its result at first, so we consulted the auther of our basic paper. Luckily,the idea also received the author's support.[1]

Design

We selected BBa_J23108 from constitutive promoter family and added a 5'UTR sequence that we choose from the paper mentioned above thereafter. Fluorescent protein is used to characterize whether or not the 5'UTR has an enhanced effect on the promoter.

Proof of concept

We constructed a circuit devices with a commonly used red fluorescent protein and randomly selected the promoter and the terminator. The difference is that UTRrpsT is added to a loop and the fluorescence intensity is measured using a microplate reader and plotted. We found that the fluorescence intensity at 6h increased to 1.5 fold. This indicates that UTRrpsT has the capability to function as a generic enhancement module.（Error bars represent standard deviation of three biological replicates.）

Future work

According to the analysis of data, we may draw a conclusion that the part can impove the expression of our target protein (mRFP). In the future, we can change different reporters to detect whether different reporters may have the same result---that their expression will all be improve. If so, we can make sure that the part is universal for improving all proteins and we can utilize this provement to increase the expression of mSA, which improve the probability of binding to yeast successfully and strengthen the binding. Further, we might offer help to other teams.

Reference

[1]Zhou, Shenghu, et al. "Obtaining a Panel of Cascade Promoter-5′-UTR Complexesin Escherichia coli." Acs Synthetic Biology 6.6(2017)



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