Team:Northwestern/Project

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<div id= "header">
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</br>
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<p id = "title"> Northwestern iGEM </p>  
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<h1> Project At a Glance </h1>
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<!-- <p id = "warning"> The site is currently under construction. The page will be pretty later. I promise!</p> -->
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      <p class="tab">The pH of the mouth drops several times per day after mealtimes, resulting in severe tooth decay over time if not addressed. One way to combat this problem is to engineer the microbiome of the mouth to respond dynamically to changes in pH. To enable this, here we present the development of a novel dual-state promoter for the detection of and response to pH fluctuations in the mouth.</p>
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<ul>
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<p class="tab"> This novel transcriptional regulation element will be capable of having both constitutive and inducible activity. This is achieved by linking a constitutive promoter downstream a pH-inducible promoter. We plan to characterize our dual-state promoter by first linking single promoters upstream green fluorescent protein (GFP) and assess the transcriptional activity via fluorescence assays. This will be followed by fluorescence assays on the dual-state promoter.</p>
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<li><a href="https://2013.igem.org/Team:Northwestern">Home</a></li>
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<p> From the construction of a dual-state promoter, we hypothesize three outcomes: </p>
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<li><a href="https://2013.igem.org/Team:Northwestern/Team">Team</a>
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<p>
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<ul>
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<ul>
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<li><a href="https://igem.org/Team.cgi?year=2013&team_name=Northwestern">Official Profile</a></li>
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          <li> The pH-inducible promoters will demonstrate minimal activity at a neutral pH, with an intense activity as the pH nears and crossed the cavity formation threshold, at pH 5.5</li>
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<li><a href="#">Meet Us!</a></li>
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        <li>The constitutive promoters will have a constant activity, independent of pH. </li>
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<li><a href="https://2013.igem.org/Team:Northwestern/Attributions">Attribution</a></li>
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        <li>The dual-state promoter construct will exhibit the combined effect of both promoters, performing at a constant basal expression as well as at an elevated level when prompted with a drop in pH. </li>
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<li><a href="https://2013.igem.org/Team:Northwestern/Project">Project</a>
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<p>This technology should have future applications in any system requiring pH detection and response.</p>
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<ul>
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<li><a href="https://2013.igem.org/Team:Northwestern/Parts">Parts Submitted</a></li>
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<li><a href="https://2013.igem.org/Team:Northwestern/Notebook">Notebook</a></li>
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<li><a href="#">Description</a></li>
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<li><a href="https://2013.igem.org/Team:Northwestern/Modeling">Modeling</a></li>
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<li><a href= "https://2013.igem.org/Team:Northwestern/Safety"> Safety </a></li>
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<h1> overall project </h1>
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<h1> Background </h1>
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<p> Oral health is one of the most overlooked aspects of health care in the world. According to the World Health Organization, 60-90% of children worldwide have dental cavities, while they are present in nearly 100% of adults1. Tooth decay and cavity development are a direct result of the plaque that accumulates in the mouth after meals. The plaque is a biofilm composed of a number of different types of bacteria native to the oral biome2. These bacteria, in particular Streptococcus mutans, secrete lactic acid as a result of sucrose metabolism, and the resulting drop in pH causes demineralization of the enamel3. Since the plaque traps the lactic acid on the surface of the teeth this pH drop takes place in direct contact with the enamel at a significant concentration. This pH drop is most prevalent directly following mealtimes. The threshold pH at which demineralization occurs is 5.5. The figure below shows that the surface of the enamel can spend nearly 5 hours a day exposed to pH below the demineralization threshold, thus this is a serious problem that provides motivation for this research. </p>
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    <div class=icons>
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      <a href="https://2013.igem.org/Team:Northwestern/problem"><h2><img src="https://static.igem.org/mediawiki/2013/thumb/2/2c/Summary-of-problem.gif/600px-Summary-of-problem.gif" height="60" width="45"/>Go to Summary of Problem</h2></a>
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    <img src="https://static.igem.org/mediawiki/2013/f/fe/Mouth_pH.png" />
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     <p>The Stephan Curve4, depicting pH fluctuation in the mouth over 24 hours.</p>
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      <a href="https://2013.igem.org/Team:Northwestern/detectpH"><h2><img src="https://static.igem.org/mediawiki/2013/thumb/1/1e/Ph-image.gif/120px-Ph-image.gif" height="60" width="40"/>Go to Detection of pH Levels</h2></a>
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<a href="https://2013.igem.org/Team:Northwestern/dualstate"><h2><img src="https://static.igem.org/mediawiki/2013/thumb/1/14/Dual-state-image.gif/600px-Dual-state-image.gif" height="60" width="40"/>Go to Dual State Promoter</h2></a>
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      <a href="https://2013.igem.org/Team:Northwestern/Results/PreFluorescence"><h2><img src="https://static.igem.org/mediawiki/2013/e/e0/MOdelling-or-results.gif" height="60" width="40"/>Go to Experiments and Results</h2></a>  
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<a href="https://2013.igem.org/Team:Northwestern/methods"><h2><img src="https://static.igem.org/mediawiki/2013/thumb/b/bf/Results-willie.gif/389px-Results-willie.gif" height="60" width="40"/>Go to Methods and Protocol</h2></a></div>
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<a href="https://2013.igem.org/Team:Northwestern/future"><h2><img src="https://static.igem.org/mediawiki/2013/1/15/Future-directions-icon.gif" height="60" width="40"/>Future Directions</h2></a>
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<h1> project details </h1>
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        <h2> experiments </h2>
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        <h2> results </h2>
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Latest revision as of 04:07, 28 September 2013


Project At a Glance

The pH of the mouth drops several times per day after mealtimes, resulting in severe tooth decay over time if not addressed. One way to combat this problem is to engineer the microbiome of the mouth to respond dynamically to changes in pH. To enable this, here we present the development of a novel dual-state promoter for the detection of and response to pH fluctuations in the mouth.

This novel transcriptional regulation element will be capable of having both constitutive and inducible activity. This is achieved by linking a constitutive promoter downstream a pH-inducible promoter. We plan to characterize our dual-state promoter by first linking single promoters upstream green fluorescent protein (GFP) and assess the transcriptional activity via fluorescence assays. This will be followed by fluorescence assays on the dual-state promoter.

From the construction of a dual-state promoter, we hypothesize three outcomes:

  • The pH-inducible promoters will demonstrate minimal activity at a neutral pH, with an intense activity as the pH nears and crossed the cavity formation threshold, at pH 5.5
  • The constitutive promoters will have a constant activity, independent of pH.
  • The dual-state promoter construct will exhibit the combined effect of both promoters, performing at a constant basal expression as well as at an elevated level when prompted with a drop in pH.

This technology should have future applications in any system requiring pH detection and response.