http://2011.igem.org/wiki/index.php?title=Special:Contributions/Eksch9&feed=atom&limit=50&target=Eksch9&year=&month=2011.igem.org - User contributions [en]2024-03-29T08:23:08ZFrom 2011.igem.orgMediaWiki 1.16.0http://2011.igem.org/Team:Missouri_Miners/TeamTeam:Missouri Miners/Team2011-09-29T04:06:16Z<p>Eksch9: </p>
<hr />
<div>{{OrganizationS&T}}<br />
<br />
<div style="width: 740px; margin: 30px; padding: 10px; background-color: #000000; color: silver; font-size: medium"><br />
<h1>Missouri Miners Team</h1><br />
<br /><br />
<br />
<h3 style="color: silver; font-weight: bold; font-size: large">Instructor:</h3><br />
<div style="margin-left: 10px;"><br />
<h4 style="color: #42C555; font-weight: bold; font-size: large">Dr. David Westenberg</h4><br />
<p style="text-indent: 0px">Associate Professor of Biological Sciences</p><br />
<br><br />
[[File:Djwesten.jpg|link=http://www.reddit.com/r/biology]]<br />
<br />
<p style="text-indent: 0px; font-weight: bold ">Education:</p><br />
<p>University of CA - Los Angeles, Doctor of Philosophy, 1991</p><br />
<br />
<p style="text-indent: 0px; font-weight: bold ">Research Interests:</p><br />
*Regulation of the bradyrhizobium japonicum sdh operon encoding succinate dehydrogenase<br />
*Evidence for AHL autoinducer production by the soybean symbiont bradyrhizobium japonicum<br />
*High efficiency separation of microbial aggregates using capillary electrophoresis<br />
*separating microbes in the manner of molecules. 1. Capillary electrokinetic approaches<br />
*Succinate dehydrogenase (Sdh) from Bradyrhizobium japonicum is closely related to mitochondrial Sdh<br />
</div><br />
<br />
<h3 style="color: silver; font-weight: bold; font-size: large">Advisor:</h3><br />
<div style="margin-left: 10px;"><br />
<h4 style="color: #42C555; font-weight: bold; font-size: large">Dr. Katie Shannon</h4><br />
<p style="text-indent: 0px">Associate Professor of Biological Sciences</p><br />
<br><br />
[[File:shannonk.jpg]]<br />
<br />
<p style="text-indent: 0px; font-weight: bold ">Education:</p><br />
<p>Harvard Medical School, Doctor of Philosophy, 2000</p><br />
<br />
<p style="text-indent: 0px; font-weight: bold ">Research Interests:</p><br />
*Cytokinesis<br />
*Cell Cycle <br />
*Cytoskeleton <br />
*Nanoparticle uptake and transport<br />
</div><br />
<br />
<h3 style="color: silver; font-weight: bold; font-size: large">Students:</h3><br />
{|<br />
|-<br />
! style="width:240px" | <br />
! style="width:240px" | <br />
! style="width:240px" | <br />
|-<br />
| style="color:#42C555; font-weight: bold; font-size: large" | Erica Shannon<br />
| style="color:#42C555; font-weight: bold; font-size: large" | Amanda Foster<br />
| style="color:#42C555; font-weight: bold; font-size: large" | April Pummill<br />
|- <br />
| [[File:MST_Erica.png|180x240px]]<br />
| [[File:Amanda.jpg|180x240px]]<br />
| [[File:April.jpg|180x240px]]<br />
|-<br />
| style="color: silver" | President <br />
| style="color: silver" | Vice-President<br />
| style="color: silver" | Public Relations<br />
|-<br />
| style="color: silver" | Major: Biological Sciences<br />
| style="color: silver" | Major: Biology & BiochemE<br />
| style="color: silver" | Major: BiochemE<br />
|-<br />
| style="color:#42C555; font-weight: bold; font-size: large" | Blythe Ferriere<br />
| style="color:#42C555; font-weight: bold; font-size: large" | Brice Curtin<br />
| style="color:#42C555; font-weight: bold; font-size: large" | Lou Harmon<br />
|- <br />
| [[File:Blythe.jpg|180x240px]]<br />
| [[File:BriceCurtin.jpg|180x240px]]<br />
| [[File:LouHarmon.jpg|180x240px]]<br />
|-<br />
| style="color: silver" | Treasurer<br />
| style="color: silver" | Secretary<br />
| style="color: silver" | Webmaster<br />
|-<br />
| style="color: silver" | Major: Chemical Engineering<br />
| style="color: silver" | Major: Chemistry<br />
| style="color: silver" | Major: Computer Science<br />
|-<br />
| style="color:#42C555; font-weight: bold; font-size: large" | Alie Abele <br />
| style="color:#42C555; font-weight: bold; font-size: large" | David Pohlman<br />
| style="color:#42C555; font-weight: bold; font-size: large" | Erica McFarland<br />
|- <br />
| [[File:Alie.jpg|180x240px]]<br />
| [[File:DavidPohlman.jpg|180x240px]]<br />
| [[File:Erica.jpg|180x240px]]<br />
|-<br />
| style="color: silver" | Major: <br />
| style="color: silver" | Lab Manager<br />
| style="color: silver" | Major:<br />
|-<br />
| style="color: silver" | <br />
| style="color: silver" | Major: BiochemE<br />
| style="color: silver" | <br />
|-<br />
| style="color:#42C555; font-weight: bold; font-size: large" | Hannah Frye<br />
| style="color:#42C555; font-weight: bold; font-size: large" | Beth Wilkins<br />
| style="color:#42C555; font-weight: bold; font-size: large" | Emily Puleo<br />
|- <br />
| [[File:HannahFrye.jpg|180x240px]]<br />
| [[File:ElizabethWilkins.jpg|180x240px]]<br />
| [[File:EmilyPuleo.png|180x240px]]<br />
|-<br />
| style="color: silver" | Major: Biochemistry<br />
| style="color: silver" | Major: Biology & Chemistry<br />
| style="color: silver" | Major: Biochemistry<br />
|- <br />
| style="color:#42C555; font-weight: bold; font-size: large" | Scott Hack <br />
| style="color:#42C555; font-weight: bold; font-size: large" | Logan Sauerbrei<br />
| style="color:#42C555; font-weight: bold; font-size: large" | Chester Gregg<br />
|- <br />
| [[File:ScottHack.jpg|180x240px]]<br />
| [[File:Logan.jpg|180x240px]]<br />
| [[File:ChesterGregg.jpg|180x240px]]<br />
|-<br />
| style="color: silver" | Major: Biology & EnvE <br />
| style="color: silver" | Major: Biological Sciences<br />
| style="color: silver" | Major: CompSci & Physics<br />
|- <br />
| style="color:#42C555; font-weight: bold; font-size: large" | Amber Kreps<br />
| style="color:#42C555; font-weight: bold; font-size: large" | Christy Kwon <br />
| style="color:#42C555; font-weight: bold; font-size: large" | Gavin Pringle<br />
|- <br />
| [[File:AmberKreps.jpg|180x240px]]<br />
| [[File:ChristyKwon.jpg|180x240px]]<br />
| [[File:GavinPringle.jpg|180x240px]]<br />
|-<br />
| style="color: silver" | Major: Biological Sciences<br />
| style="color: silver" | Major: <br />
| style="color: silver" | Major: <br />
|- <br />
| style="color:#42C555; font-weight: bold; font-size: large" | Greg Niles<br />
| style="color:#42C555; font-weight: bold; font-size: large" | Tyler Robinson<br />
| style="color:#42C555; font-weight: bold; font-size: large" | Thomas Congdon<br />
|- <br />
| [[File:GregNiles.jpg|180x240px]]<br />
| [[File:TylerRobinson.jpg|180x240px]]<br />
| [[File:ThomasCongdon.jpg|180x240px]]<br />
| <br />
|-<br />
| style="color: silver" | Major: Mechanical Eng.<br />
| style="color: silver" | Major: <br />
| style="color: silver" | Major: BiomedE<br />
|}<br />
</div></div>Eksch9http://2011.igem.org/Team:Missouri_Miners/DataTeam:Missouri Miners/Data2011-09-29T03:37:32Z<p>Eksch9: </p>
<hr />
<div>{{OrganizationS&T}}<br />
<br />
<html><br />
<div style="width: 740px; margin: 30px; padding: 10px; background-color: #000000; color: silver; font-size: medium"><br />
<h1>Data</h1><br />
<br />
<br />
<br />
<p /> Our part consists of an OmpR binding region that regulates our reporter gene, eyfp. The OmpR binding region is the ompC operator, which has three OmpR binding sites. Our part is regulated by the EnvZ-OmpR two-component regulatory system. EnvZ is an osmolarity sensor on the inner membrane and phosphorylates OmpR in the presence of high osmolarity. When phosphorylated, OmpR binds to the ompC operator and recruits RNAP to transcribe the downstream reporter gene. Fluorescence intensity due to the production of eYfp is a measure of the system's activity. When all three binding sites are occupied by phosphorylated OmpR proteins, RNAP cannot bind to the DNA and transcribe downstream. This part can be used as an osmolarity indicator. Our team used this part as a glucose sensor because the presence of glucose changes osmolarity. Different concentrations of glucose produce different, quantifiable fluorescence intensities.<br/><br/><br />
<br />
<br />
<br />
<h3>Sequence Verification</h3><br />
This part has been sequence verified and works as expected. <br/><br/><br />
<br />
<br />
<h3>Fluorescence Wavelengths</h3><br />
*'''Excitation max''' - 485nm<br/><br />
*'''Emission max''' - 520nm<br/><br/><br />
<br />
<br />
<br />
<h3>Growth Conditions</h3><br />
*'''Growth Temperature and Rate''' - E. colistrain DH5alphaused as Chassis, standard growth conditions apply <br/><br />
*'''Growth Media''' - Nutrient Broth or diluted LB media, since OmpR is an osmolarity receptor the cells must be grown in a low osmolarity media to control for compounding factors. To express Fluorescence it is recommended that cultures are grown in liquid media. Colonies growing on plates cannot detect osmolarity and will not activate EnvZ. <br/><br/><br />
<br />
If cells are grown in a high osmolarity media, such as LB broth, the background fluorescence will be higher than an induced glucose response. This is because the system is already primed from the high osmolarity of the media. When additional osmolites are added, the system will down regulate eyfp due to the presence of too many phosphorylated OmpR proteins. Below is a dose response in the presence of LB media. Note how glucose, in these conditions, has an inhibitory effect on the system. <br/><br/><br />
</html><br />
[[Image:Grown in LB.jpg]]<br />
<html><br />
<br/><br/><br />
<br />
<h3>Dose Response</h3><br />
To control for compounding variables, the cells assayed were grown in 0.5X LB media. This reduced the background osmolarity and allowed our team to measure a valid response. In the graph you can see that the system fluoresces over background at 1% Glucose and then drops back to background at 5% Glucose. The increase at 1% is due to positive regulation by the EnvZ-OmpR system. 5% glucose is a high enough concentration to induce downregulation of the system. Too many phosphorylated OmpR proteins bind the ompC promoter. When all three binding sites are filled RNAP cannot transcribe downstream and fluorescence decreases. <br/><br/><br />
<br />
<br />
<img src="https://static.igem.org/mediawiki/2011/4/48/Dose_response_1.jpg" alt="Dose Response"> <br/><br/><br />
<br />
<h3>Time Trial</h3><br />
The graph below shows time trials of the system. These time trials were run in the presence 0%, 1%, and 8% glucose. We chose to do this so we could gather more comprehensive data. The first conclusion we can draw from this graph is that the response is fairly consistent over time (there is no peak in fluorescence associated with a specific time point). Also, for each time point between 0.5-4.5 hrs, the curve of the dose response remains consistent. This graph also shows that the assay is not accurate after 4.5 hours. The data beyond 4.5 hours becomes random and inconsistent.<br/><br/><br />
<br />
<div style="text-align:center;"><br />
<img src="https://static.igem.org/mediawiki/2011/f/fe/Time_Trials_2.jpg" alt="Time Trials" align="top" style="width:720px"><br/><br/><br />
</div><br />
</div><br />
</html></div>Eksch9http://2011.igem.org/Team:Missouri_Miners/ProjectTeam:Missouri Miners/Project2011-09-29T03:26:36Z<p>Eksch9: </p>
<hr />
<div>{{OrganizationS&T}}<br />
<br />
<div id="box" style="position: relative; width: 740px; margin-left: 30px; top:-10px; padding: 5px; background-color: #000000;"> <br />
<div id="template" style="font-weight: regular; font-size: medium; color: silver; padding: 5px;"><br />
<h1>Project </h1><br /><br />
<br />
<h3>Abstract</h3><br />
<p>In the bodies of people with diabetes, the ability to recognize and respond to glucose concentrations in the blood has been compromised. As a result, glucose accumulates to dangerous levels. High blood glucose concentrations can cause irreversible damage to critical organs, impairing their functionality. With parts from the iGEM registry, our team created a glucose-controlled promoter linked to a yellow fluorescence production gene in E. coli. The concentrations of glucose to which the promoter responds can be determined. Once the concentration is known, the promoter can be mutated so that it will be activated by varying concentrations of glucose and be used as a glucose sensor for people with diabetes. In the future, an insulin gene could be added to this system for use in insulin pumps, where specific glucose levels trigger insulin production in E. coli.</p><br />
<br /><br />
<h3>Our System Overview</h3><br />
[[File:IGEM System.JPG|thumbnail|right|360px|Two Component Regulatory System Pathway]]<br />
<p>We submitted two biobricks to the registry: an [http://partsregistry.org/wiki/index.php?title=Part:BBa_K621000| intermediate part] with a ribosome binding site and eYFP, and the [http://partsregistry.org/wiki/index.php?title=Part:BBa_K621001| intermediate plus the ompC operator]. The ompC operator has three binding sites for phosphorylated OmpR. OmpR and EnvZ work together in a two-component regulatory system as shown in the diagram below. </p><br />
<br /><br />
[[File:IGEM binding.JPG|thumbnail|right|360px|When one or two OmpR bind, RNAP transcribes the gene.]]<br />
[[File:IGEM binding2.JPG|thumb|right|360px|When all OmpR binding sites are occupied, RNAP can't bind.]]<br />
<p>EnvZ is an inner membrane protein that senses osmolarity. Phosphorlyation of OmpR by EnvZ positively correlates with the osmolarity of the system. Phosphorylated OmpR (OmpR-P) can bind to the three sites on the ompC operator. When one or two of the binding sites are occupied by OmpR-P, RNA polymerase is recruited to begin downstream transcription of the reporter gene, eYFP. However, when all three OmpR binding sites are occupied by OmpR-P, RNA polymerase cannot bind, the reporter gene can no longer be produced, and therefore the system is inhibited. In summary, as osmolarity increases from very low levels, the fluorescence produced by the system also increases until the system reaches a threshold osmolarity. Once the system reaches the threshold, the fluorescence will decrease with increasing osmolarity due to the inherent down-regulation of the system. The activity of the system can be quantified using the fluorescence of the cells because the two-component regulatory system of EnvZ and OmpR regulates transcription of the eYFP gene, dictating the level of fluorescence.</p><br />
<div style="height: 795px"><br />
<h3>Making Our Parts</h3><br />
[[File:Igemmodel3.jpg|right|400px]]<br />
<p>We started with three parts from the iGEM registry:<br />
*BBa_B0032: RBS-3, a medium-strength ribosome binding site<br />
*BBa_E0030: eYFP gene, codes for yellow fluorescence<br />
*BBa_R0082: omp-c operon, contains three binding sites for phosphorylated OmpR</p>To build our part our team performed restriction digests and ligations as indicated by the figure to the right .<br />
<br />
</div><br />
<br /><br />
<h3>Testing</h3><br />
<p>To measure fluorescence our team used a 96-well plate reader. Overnight cultures of bacteria were grown in 0.5X LB media and then aliquoted into 1.5ml tubes. Serial dilutions of the desired glucose concentrations were made. Glucose was diluted using 0.5X LB media so that the concentration of LB remained constant. The glucose was administered to the cells in a fashion that did not dilute the cell densities in the cultures. Samples of each treatment were added to a 96-well plate and were imaged using the plate reader. See [[Team:Missouri_Miners/data| data]] page for results. </p><br />
<br /><br />
<h3>Next Steps</h3><br />
<p>In the future, our system has applications in glucose sensing devices. For our system to be sensitive to different changes in osmolarity the OmpC operon will need to be mutated. The mutagenesis process begins with transforming our system into a mutagenic strain of E. coli, in which, the cells have no proof reading capability during DNA replication. In these cells the rate of mutation is increased by 1000x. Using this process mutant strains could be produced, sequenced, and characterized. Possible applications of a characterized system include economical glucose testing strips, a wider range of test strip availability, and precursors to advanced insulin pump systems. </p><br />
</div><br />
</div></div>Eksch9http://2011.igem.org/Team:Missouri_Miners/ProjectTeam:Missouri Miners/Project2011-09-29T03:19:44Z<p>Eksch9: </p>
<hr />
<div>{{OrganizationS&T}}<br />
<br />
<div id="box" style="position: relative; width: 740px; margin-left: 30px; top:-10px; padding: 5px; background-color: #000000;"> <br />
<div id="template" style="font-weight: regular; font-size: medium; color: silver; padding: 5px;"><br />
<h1>Project </h1><br /><br />
<br />
<h3>Abstract</h3><br />
<p>In the bodies of people with diabetes, the ability to recognize and respond to glucose concentrations in the blood has been compromised. As a result, glucose accumulates to dangerous levels. High blood glucose concentrations can cause irreversible damage to critical organs, impairing their functionality. With parts from the iGEM registry, our team created a glucose-controlled promoter linked to a yellow fluorescence production gene in E. coli. The concentrations of glucose to which the promoter responds can be determined. Once the concentration is known, the promoter can be mutated so that it will be activated by varying concentrations of glucose and be used as a glucose sensor for people with diabetes. In the future, an insulin gene could be added to this system for use in insulin pumps, where specific glucose levels trigger insulin production in E. coli.</p><br />
<br /><br />
<h3>Our System Overview</h3><br />
<p>We submitted two biobricks to the registry: an intermediate part (http://partsregistry.org/wiki/index.php?title=Part:BBa_K621000) with a ribosome binding site and eYFP, and the intermediate plus the ompC operator (http://partsregistry.org/wiki/index.php?title=Part:BBa_K621001). The ompC operator has three binding sites for phosphorylated OmpR. OmpR and EnvZ work together in a two-component regulatory system as shown in the diagram below. </p><br />
<br /><br />
[[File:IGEM binding.JPG|thumbnail|right|360px|When one or two OmpR bind, RNAP transcribes the gene.]]<br />
[[File:IGEM binding2.JPG|thumb|right|360px|When all OmpR binding sites are occupied, RNAP can't bind.]]<br />
<p>EnvZ is an inner membrane protein that senses osmolarity. Phosphorlyation of OmpR by EnvZ positively correlates with the osmolarity of the system. Phosphorylated OmpR (OmpR-P) can bind to the three sites on the ompC operator. When one or two of the binding sites are occupied by OmpR-P, RNA polymerase is recruited to begin downstream transcription of the reporter gene, eYFP. However, when all three OmpR binding sites are occupied by OmpR-P, RNA polymerase cannot bind, the reporter gene can no longer be produced, and therefore the system is inhibited. In summary, as osmolarity increases from very low levels, the fluorescence produced by the system also increases until the system reaches a threshold osmolarity. Once the system reaches the threshold, the fluorescence will decrease with increasing osmolarity due to the inherent down-regulation of the system. The activity of the system can be quantified using the fluorescence of the cells because the two-component regulatory system of EnvZ and OmpR regulates transcription of the eYFP gene, dictating the level of fluorescence.</p><br />
<div style="height: 795px"><br />
<h3>Making Our Parts</h3><br />
[[File:Igemmodel3.jpg|right|400px]]<br />
<p>We started with three parts from the iGEM registry:<br />
*BBa_B0032: RBS-3, a medium-strength ribosome binding site<br />
*BBa_E0030: eYFP gene, codes for yellow fluorescence<br />
*BBa_R0082: omp-c operon, contains three binding sites for phosphorylated OmpR</p>To build our part our team performed restriction digests and ligations as indicated by the figure to the right .<br />
<br />
</div><br />
<br /><br />
<h3>Testing</h3><br />
<p>To measure fluorescence our team used a 96-well plate reader. Overnight cultures of bacteria were grown in 0.5X LB media and then aliquoted into 1.5ml tubes. Serial dilutions of the desired glucose concentrations were made. Glucose was diluted using 0.5X LB media so that the concentration of LB remained constant. The glucose was administered to the cells in a fashion that did not dilute the cell densities in the cultures. Samples of each treatment were added to a 96-well plate and were imaged using the plate reader. See [[Team:Missouri_Miners/data| data]] page for results. </p><br />
<br /><br />
<h3>Next Steps</h3><br />
<p>In the future, our system has applications in glucose sensing devices. For our system to be sensitive to different changes in osmolarity the OmpC operon will need to be mutated. The mutagenesis process begins with transforming our system into a mutagenic strain of E. coli, in which, the cells have no proof reading capability during DNA replication. In these cells the rate of mutation is increased by 1000x. Using this process mutant strains could be produced, sequenced, and characterized. Possible applications of a characterized system include economical glucose testing strips, a wider range of test strip availability, and precursors to advanced insulin pump systems. </p><br />
</div><br />
</div></div>Eksch9http://2011.igem.org/Template:OrganizationS%26TTemplate:OrganizationS&T2011-09-29T03:13:49Z<p>Eksch9: </p>
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numTweets: 1,<br />
loaderText: "Loading tweet...",<br />
slideIn: true,<br />
slideDuration: 750,<br />
showHeading: true,<br />
headingText: "Latest Tweets",<br />
showProfileLink: true,<br />
showTimestamp: true<br />
};<br />
<br />
var o = $.extend({}, $.fn.getTwitter.defaults, options);<br />
<br />
return this.each(function() {<br />
var c = $(this);<br />
<br />
// hide container element, remove alternative content, and add class<br />
c.hide().empty().addClass("twitted");<br />
<br />
// add heading to container element<br />
if (o.showHeading) {<br />
c.append("<h2>"+o.headingText+"</h2>");<br />
}<br />
<br />
// add Twitter profile link to container element<br />
if (o.showProfileLink) {<br />
var profileLinkHTML = "<p class=\"profileLink\"><a href=\"http://twitter.com/"+o.userName+"\">http://twitter.com/"+o.userName+"</a></p>";<br />
c.append(profileLinkHTML);<br />
}<br />
<br />
// add twitter list to container element<br />
var twitterListHTML = "<ul id=\"twitter_update_list\"><li></li></ul>";<br />
c.append(twitterListHTML);<br />
<br />
var tl = $("#twitter_update_list");<br />
<br />
// hide twitter list<br />
tl.hide();<br />
<br />
// add preLoader to container element<br />
var preLoaderHTML = $("<p class=\"preLoader\">"+o.loaderText+"</p>");<br />
c.append(preLoaderHTML);<br />
<br />
// show container element<br />
c.show();<br />
<br />
$.getScript("http://twitter.com/javascripts/blogger.js");<br />
$.getScript("http://twitter.com/statuses/user_timeline/"+o.userName+".json?callback=twitterCallback2&count="+o.numTweets, function() {<br />
// remove preLoader from container element<br />
$(preLoaderHTML).remove();<br />
<br />
// remove timestamp and move to title of list item<br />
if (!o.showTimestamp) {<br />
tl.find("li").each(function() {<br />
var timestampHTML = $(this).children("a");<br />
var timestamp = timestampHTML.html();<br />
timestampHTML.remove();<br />
$(this).attr("title", timestamp);<br />
});<br />
}<br />
<br />
// show twitter list<br />
if (o.slideIn) {<br />
var tlHeight = tl.data("originalHeight");<br />
<br />
// get the original height<br />
if (!tlHeight) {<br />
tlHeight = tl.show().height();<br />
tl.data("originalHeight", tlHeight);<br />
tl.hide().css({height: 0});<br />
}<br />
<br />
tl.show().animate({height: tlHeight}, o.slideDuration);<br />
}<br />
else {<br />
tl.show();<br />
}<br />
<br />
// add unique class to first list item<br />
tl.find("li:first").addClass("firstTweet");<br />
<br />
// add unique class to last list item<br />
tl.find("li:last").addClass("lastTweet");<br />
});<br />
});<br />
};<br />
})(jQuery);<br />
</script><br />
<script type="text/javascript"><br />
/*<br />
* jQuery Easing v1.3 - http://gsgd.co.uk/sandbox/jquery/easing/<br />
*<br />
* Uses the built in easing capabilities added In jQuery 1.1<br />
* to offer multiple easing options<br />
*<br />
* TERMS OF USE - jQuery Easing<br />
* <br />
* Open source under the BSD License. <br />
* <br />
* Copyright c 2008 George McGinley Smith<br />
* All rights reserved.<br />
* <br />
* Redistribution and use in source and binary forms, with or without modification, <br />
* are permitted provided that the following conditions are met:<br />
* <br />
* Redistributions of source code must retain the above copyright notice, this list of <br />
* conditions and the following disclaimer.<br />
* Redistributions in binary form must reproduce the above copyright notice, this list <br />
* of conditions and the following disclaimer in the documentation and/or other materials <br />
* provided with the distribution.<br />
* <br />
* Neither the name of the author nor the names of contributors may be used to endorse <br />
* or promote products derived from this software without specific prior written permission.<br />
* <br />
* THIS SOFTWARE IS PROVIDED BY THE COPYRIGHT HOLDERS AND CONTRIBUTORS "AS IS" AND ANY <br />
* EXPRESS OR IMPLIED WARRANTIES, INCLUDING, BUT NOT LIMITED TO, THE IMPLIED WARRANTIES OF<br />
* MERCHANTABILITY AND FITNESS FOR A PARTICULAR PURPOSE ARE DISCLAIMED. IN NO EVENT SHALL THE<br />
* COPYRIGHT OWNER OR CONTRIBUTORS BE LIABLE FOR ANY DIRECT, INDIRECT, INCIDENTAL, SPECIAL,<br />
* EXEMPLARY, OR CONSEQUENTIAL DAMAGES (INCLUDING, BUT NOT LIMITED TO, PROCUREMENT OF SUBSTITUTE<br />
* GOODS OR SERVICES; LOSS OF USE, DATA, OR PROFITS; OR BUSINESS INTERRUPTION) HOWEVER CAUSED <br />
* AND ON ANY THEORY OF LIABILITY, WHETHER IN CONTRACT, STRICT LIABILITY, OR TORT (INCLUDING<br />
* NEGLIGENCE OR OTHERWISE) ARISING IN ANY WAY OUT OF THE USE OF THIS SOFTWARE, EVEN IF ADVISED <br />
* OF THE POSSIBILITY OF SUCH DAMAGE. <br />
*<br />
*/<br />
<br />
// t: current time, b: begInnIng value, c: change In value, d: duration<br />
jQuery.easing['jswing'] = jQuery.easing['swing'];<br />
<br />
jQuery.extend( jQuery.easing,<br />
{<br />
def: 'easeOutQuad',<br />
swing: function (x, t, b, c, d) {<br />
//alert(jQuery.easing.default);<br />
return jQuery.easing[jQuery.easing.def](x, t, b, c, d);<br />
},<br />
easeInQuad: function (x, t, b, c, d) {<br />
return c*(t/=d)*t + b;<br />
},<br />
easeOutQuad: function (x, t, b, c, d) {<br />
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},<br />
easeInOutQuad: function (x, t, b, c, d) {<br />
if ((t/=d/2) < 1) return c/2*t*t + b;<br />
return -c/2 * ((--t)*(t-2) - 1) + b;<br />
},<br />
easeInCubic: function (x, t, b, c, d) {<br />
return c*(t/=d)*t*t + b;<br />
},<br />
easeOutCubic: function (x, t, b, c, d) {<br />
return c*((t=t/d-1)*t*t + 1) + b;<br />
},<br />
easeInOutCubic: function (x, t, b, c, d) {<br />
if ((t/=d/2) < 1) return c/2*t*t*t + b;<br />
return c/2*((t-=2)*t*t + 2) + b;<br />
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easeInQuart: function (x, t, b, c, d) {<br />
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easeOutQuart: function (x, t, b, c, d) {<br />
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easeInOutQuart: function (x, t, b, c, d) {<br />
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return -c/2 * ((t-=2)*t*t*t - 2) + b;<br />
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easeInQuint: function (x, t, b, c, d) {<br />
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easeOutQuint: function (x, t, b, c, d) {<br />
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easeInSine: function (x, t, b, c, d) {<br />
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easeOutSine: function (x, t, b, c, d) {<br />
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easeInOutSine: function (x, t, b, c, d) {<br />
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easeInExpo: function (x, t, b, c, d) {<br />
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easeOutExpo: function (x, t, b, c, d) {<br />
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easeInOutExpo: function (x, t, b, c, d) {<br />
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if (t==d) return b+c;<br />
if ((t/=d/2) < 1) return c/2 * Math.pow(2, 10 * (t - 1)) + b;<br />
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easeInCirc: function (x, t, b, c, d) {<br />
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},<br />
easeOutCirc: function (x, t, b, c, d) {<br />
return c * Math.sqrt(1 - (t=t/d-1)*t) + b;<br />
},<br />
easeInOutCirc: function (x, t, b, c, d) {<br />
if ((t/=d/2) < 1) return -c/2 * (Math.sqrt(1 - t*t) - 1) + b;<br />
return c/2 * (Math.sqrt(1 - (t-=2)*t) + 1) + b;<br />
},<br />
easeInElastic: function (x, t, b, c, d) {<br />
var s=1.70158;var p=0;var a=c;<br />
if (t==0) return b; if ((t/=d)==1) return b+c; if (!p) p=d*.3;<br />
if (a < Math.abs(c)) { a=c; var s=p/4; }<br />
else var s = p/(2*Math.PI) * Math.asin (c/a);<br />
return -(a*Math.pow(2,10*(t-=1)) * Math.sin( (t*d-s)*(2*Math.PI)/p )) + b;<br />
},<br />
easeOutElastic: function (x, t, b, c, d) {<br />
var s=1.70158;var p=0;var a=c;<br />
if (t==0) return b; if ((t/=d)==1) return b+c; if (!p) p=d*.3;<br />
if (a < Math.abs(c)) { a=c; var s=p/4; }<br />
else var s = p/(2*Math.PI) * Math.asin (c/a);<br />
return a*Math.pow(2,-10*t) * Math.sin( (t*d-s)*(2*Math.PI)/p ) + c + b;<br />
},<br />
easeInOutElastic: function (x, t, b, c, d) {<br />
var s=1.70158;var p=0;var a=c;<br />
if (t==0) return b; if ((t/=d/2)==2) return b+c; if (!p) p=d*(.3*1.5);<br />
if (a < Math.abs(c)) { a=c; var s=p/4; }<br />
else var s = p/(2*Math.PI) * Math.asin (c/a);<br />
if (t < 1) return -.5*(a*Math.pow(2,10*(t-=1)) * Math.sin( (t*d-s)*(2*Math.PI)/p )) + b;<br />
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easeInBack: function (x, t, b, c, d, s) {<br />
if (s == undefined) s = 1.70158;<br />
return c*(t/=d)*t*((s+1)*t - s) + b;<br />
},<br />
easeOutBack: function (x, t, b, c, d, s) {<br />
if (s == undefined) s = 1.70158;<br />
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easeInOutBack: function (x, t, b, c, d, s) {<br />
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if ((t/=d/2) < 1) return c/2*(t*t*(((s*=(1.525))+1)*t - s)) + b;<br />
return c/2*((t-=2)*t*(((s*=(1.525))+1)*t + s) + 2) + b;<br />
},<br />
easeInBounce: function (x, t, b, c, d) {<br />
return c - jQuery.easing.easeOutBounce (x, d-t, 0, c, d) + b;<br />
},<br />
easeOutBounce: function (x, t, b, c, d) {<br />
if ((t/=d) < (1/2.75)) {<br />
return c*(7.5625*t*t) + b;<br />
} else if (t < (2/2.75)) {<br />
return c*(7.5625*(t-=(1.5/2.75))*t + .75) + b;<br />
} else if (t < (2.5/2.75)) {<br />
return c*(7.5625*(t-=(2.25/2.75))*t + .9375) + b;<br />
} else {<br />
return c*(7.5625*(t-=(2.625/2.75))*t + .984375) + b;<br />
}<br />
},<br />
easeInOutBounce: function (x, t, b, c, d) {<br />
if (t < d/2) return jQuery.easing.easeInBounce (x, t*2, 0, c, d) * .5 + b;<br />
return jQuery.easing.easeOutBounce (x, t*2-d, 0, c, d) * .5 + c*.5 + b;<br />
}<br />
});<br />
<br />
/*<br />
*<br />
* TERMS OF USE - EASING EQUATIONS<br />
* <br />
* Open source under the BSD License. <br />
* <br />
* Copyright c 2001 Robert Penner<br />
* All rights reserved.<br />
* <br />
* Redistribution and use in source and binary forms, with or without modification, <br />
* are permitted provided that the following conditions are met:<br />
* <br />
* Redistributions of source code must retain the above copyright notice, this list of <br />
* conditions and the following disclaimer.<br />
* Redistributions in binary form must reproduce the above copyright notice, this list <br />
* of conditions and the following disclaimer in the documentation and/or other materials <br />
* provided with the distribution.<br />
* <br />
* Neither the name of the author nor the names of contributors may be used to endorse <br />
* or promote products derived from this software without specific prior written permission.<br />
* <br />
* THIS SOFTWARE IS PROVIDED BY THE COPYRIGHT HOLDERS AND CONTRIBUTORS "AS IS" AND ANY <br />
* EXPRESS OR IMPLIED WARRANTIES, INCLUDING, BUT NOT LIMITED TO, THE IMPLIED WARRANTIES OF<br />
* MERCHANTABILITY AND FITNESS FOR A PARTICULAR PURPOSE ARE DISCLAIMED. IN NO EVENT SHALL THE<br />
* COPYRIGHT OWNER OR CONTRIBUTORS BE LIABLE FOR ANY DIRECT, INDIRECT, INCIDENTAL, SPECIAL,<br />
* EXEMPLARY, OR CONSEQUENTIAL DAMAGES (INCLUDING, BUT NOT LIMITED TO, PROCUREMENT OF SUBSTITUTE<br />
* GOODS OR SERVICES; LOSS OF USE, DATA, OR PROFITS; OR BUSINESS INTERRUPTION) HOWEVER CAUSED <br />
* AND ON ANY THEORY OF LIABILITY, WHETHER IN CONTRACT, STRICT LIABILITY, OR TORT (INCLUDING<br />
* NEGLIGENCE OR OTHERWISE) ARISING IN ANY WAY OUT OF THE USE OF THIS SOFTWARE, EVEN IF ADVISED <br />
* OF THE POSSIBILITY OF SUCH DAMAGE. <br />
*<br />
*/<br />
</script><br />
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<script type="text/javascript"><br />
$(function() {<br />
$('#sdt_menu > li').bind('mouseenter',function(){<br />
var $elem = $(this);<br />
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$sub_menu.hide().animate({'top':'0px','height':'0px'},0,function(){<br />
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<link href='http://fonts.googleapis.com/css?family=Nobile:regular,bold' rel='stylesheet' type='text/css'><br />
<br />
<link href='http://fonts.googleapis.com/css?family=Ubuntu:regular,bold&subset=latin' rel='stylesheet' type='text/css'><br />
<style><br />
/**<br />
* jQuery lightBox plugin<br />
* This jQuery plugin was inspired and based on Lightbox 2 by Lokesh Dhakar (http://www.huddletogether.com/projects/lightbox2/)<br />
* and adapted to me for use like a plugin from jQuery.<br />
* @name jquery-lightbox-0.5.css<br />
* @author Leandro Vieira Pinho - http://leandrovieira.com<br />
* @version 0.5<br />
* @date April 11, 2008<br />
* @category jQuery plugin<br />
* @copyright (c) 2008 Leandro Vieira Pinho (leandrovieira.com)<br />
* @license CCAttribution-ShareAlike 2.5 Brazil - http://creativecommons.org/licenses/by-sa/2.5/br/deed.en_US<br />
* @example Visit http://leandrovieira.com/projects/jquery/lightbox/ for more informations about this jQuery plugin<br />
*/<br />
#jquery-overlay {<br />
position: absolute;<br />
top: 0;<br />
left: 0;<br />
z-index: 90;<br />
width: 100%;<br />
height: 500px;<br />
}<br />
#jquery-lightbox {<br />
position: absolute;<br />
top: 0;<br />
left: 0;<br />
width: 100%;<br />
z-index: 100;<br />
text-align: center;<br />
line-height: 0;<br />
}<br />
#jquery-lightbox a img { border: none; }<br />
#lightbox-container-image-box {<br />
position: relative;<br />
background-color: #fff;<br />
width: 250px;<br />
height: 250px;<br />
margin: 0 auto;<br />
}<br />
#lightbox-container-image { padding: 10px; }<br />
#lightbox-loading {<br />
position: absolute;<br />
top: 40%;<br />
left: 0%;<br />
height: 25%;<br />
width: 100%;<br />
text-align: center;<br />
line-height: 0;<br />
}<br />
#lightbox-nav {<br />
position: absolute;<br />
top: 0;<br />
left: 0;<br />
height: 100%;<br />
width: 100%;<br />
z-index: 10;<br />
}<br />
#lightbox-container-image-box > #lightbox-nav { left: 0; }<br />
#lightbox-nav a { outline: none;}<br />
#lightbox-nav-btnPrev, #lightbox-nav-btnNext {<br />
width: 49%;<br />
height: 100%;<br />
zoom: 1;<br />
display: block;<br />
}<br />
#lightbox-nav-btnPrev { <br />
left: 0; <br />
float: left;<br />
}<br />
#lightbox-nav-btnNext { <br />
right: 0; <br />
float: right;<br />
}<br />
#lightbox-container-image-data-box {<br />
font: 10px Verdana, Helvetica, sans-serif;<br />
background-color: #fff;<br />
margin: 0 auto;<br />
line-height: 1.4em;<br />
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width: 100%;<br />
padding: 0 10px 0;<br />
}<br />
#lightbox-container-image-data {<br />
padding: 0 10px; <br />
color: #666; <br />
}<br />
#lightbox-container-image-data #lightbox-image-details { <br />
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float: left; <br />
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#lightbox-image-details-caption { font-weight: bold; }<br />
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float: right;<br />
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<style><br />
span.reference{<br />
position:fixed;<br />
left:10px;<br />
bottom:10px;<br />
font-size:12px;<br />
}<br />
span.reference a{<br />
color:#aaa;<br />
text-transform:uppercase;<br />
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span.reference a:hover{<br />
color:#ddd;<br />
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ul.sdt_menu{<br />
margin-top:150px;<br />
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<script><br />
$(document).ready(function() {<br />
$('a.lightbox').lightBox(); // Select all links with lightbox class<br />
$('a.image').each(function(idx, item) { <br />
var src = item.firstChild.src;<br />
src=src.replace(/wiki\/images\/thumb\/(.+)\/(.+)\/(.+)\/(.+)\..../,"wiki/images/$1/$2/$3");<br />
item.href=src;<br />
});<br />
$('a.image').lightBox();<br />
});<br />
</script><br />
<br />
<br />
<br />
<link rel="stylesheet" type="text/css" href="https://2011.igem.org/Team:Missouri_Miners/css?action=raw&ctype=text/css" /><br />
</head><br />
<body><br />
<div id="headerimg1" class="headerimg"></div><br />
<div id="headerimg2" class="headerimg"></div><br />
<div id="header" style="height:100px;padding: 85px 0px 0px 0px; margin-bottom:0px"><br />
<ul id="sdt_menu" class="sdt_menu"><br />
<li><br />
<a href="https://2011.igem.org/Team:Missouri_Miners"><br />
<img src="https://static.igem.org/mediawiki/2011/3/34/Homeicon.jpg" alt=""/> <br />
<span class="sdt_active"></span><br />
<span class="sdt_wrap"><br />
<span class="sdt_link">Home</span><br />
<span class="sdt_descr"></span><br />
</span><br />
</a><br />
<div class="sdt_box"><br />
<a href="https://igem.org/Team.cgi">Profile</a><br />
<a href="https://2011.igem.org/Team:Missouri_Miners/Team">Team Page</a><br />
</div><br />
</li><br />
<li><br />
<a href="https://2011.igem.org/Team:Missouri_Miners/Project"><br />
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<span class="sdt_active"></span><br />
<span class="sdt_wrap"><br />
<span class="sdt_link">Project</span><br />
<span class="sdt_descr"></span><br />
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<a href="https://2011.igem.org/Team:Missouri_Miners/Project_Background">Project Background</a><br />
<a href="https://2011.igem.org/Team:Missouri_Miners/Project">Project</a><br />
<a href="https://2011.igem.org/Team:Missouri_Miners/Parts">Parts</a><br />
<a href="https://2011.igem.org/Team:Missouri_Miners/Data">Data</a><br />
<a href="https://2011.igem.org/Team:Missouri_Miners/Notebook">Notebook</a><br />
</div><br />
</li><br />
<li><br />
<a href="https://2011.igem.org/Team:Missouri_Miners/Ethics"> <br />
<img src="http://singularityhub.com/wp-content/uploads/2009/02/gene_therapy.jpg" alt=""/> <br />
<span class="sdt_active"></span><br />
<span class="sdt_wrap"><br />
<span class="sdt_link">Ethics & Safety</span><br />
<span class="sdt_descr"></span><br />
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<a href="https://2011.igem.org/Team:Missouri_Miners/Ethics">Ethics</a><br />
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<a href="https://2011.igem.org/Team:Missouri_Miners/Organization">Organization</a><br />
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<a href="https://2011.igem.org/Team:Missouri_Miners/Papers"> Research Papers</a><br />
<a href="https://2011.igem.org/Team:Missouri_Miners/DiabetesLinks">Diabetes</a><br />
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<a href="https://2011.igem.org/Team:Missouri_Miners/InsulinPumpLinks">Insulin Pumps</a><br />
<a href="https://2011.igem.org/Team:Missouri_Miners/EthicsAndSafetyLinks">Ethics and Safety</a><br />
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</div></div>Eksch9http://2011.igem.org/Template:OrganizationS%26TTemplate:OrganizationS&T2011-09-29T03:13:00Z<p>Eksch9: </p>
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</div></div>Eksch9http://2011.igem.org/Team:Missouri_Miners/PapersTeam:Missouri Miners/Papers2011-09-29T03:08:31Z<p>Eksch9: </p>
<hr />
<div>{{OrganizationS&T}}<br />
<br />
<br />
<div id="box" style="position: relative; width: 740px; margin-left: 30px; top:-20px; padding: 5px; background-color: #000000;"> <br />
<div id="template" style="font-weight: regular; font-size: medium; color: silver; padding: 5px;"><br />
<h1>Research Papers </h1><br /><br />
<br />
<br />
[http://www.pnas.org/content/94/7/2828.full.pdf|left| Phosphorylation stimulates the cooperative DNA-binding properties of the transcription factor OmpR]<br />
<br />
[http://www.ncbi.nlm.nih.gov/pmc/articles/PMC212516/| Suppressor Mutations in rpoA Suggest that OmpR Controls Transcription by Direct Interaction with the a Subunit of RNA Polymerase]<br />
<br />
[http://www.ncbi.nlm.nih.gov/pubmed/11973328| EnvZ-OmpR Interaction and Osmoregulation in Escherichia coli*]<br />
<br />
[http://www.ncbi.nlm.nih.gov/pmc/articles/PMC297773/| Phosphorylation of OmpR by the osmosensor EnvZ modulates expression of the ompF and ompC genes in Escherichia coli]<br />
<br />
[http://www.ncbi.nlm.nih.gov/pubmed/1311295| Molecular Analysis of the Signaling Pathway between EnvZ and OmpR in Escherichia coli]<br />
<br />
[http://www.ncbi.nlm.nih.gov/pubmed/10966457| TWO-COMPONENT SIGNAL TRANSDUCTION]<br />
<br />
</div><br />
</div></div>Eksch9http://2011.igem.org/Team:Missouri_Miners/ProjectTeam:Missouri Miners/Project2011-09-29T03:00:22Z<p>Eksch9: </p>
<hr />
<div>{{OrganizationS&T}}<br />
<br />
<div id="box" style="position: relative; width: 740px; margin-left: 30px; top:-10px; padding: 5px; background-color: #000000;"> <br />
<div id="template" style="font-weight: regular; font-size: medium; color: silver; padding: 5px;"><br />
<h1>Project </h1><br /><br />
<br />
<h3>Abstract</h3><br />
<p>In the bodies of people with diabetes, the ability to recognize and respond to glucose concentrations in the blood has been compromised. As a result, glucose accumulates to dangerous levels. High blood glucose concentrations can cause irreversible damage to critical organs, impairing their functionality. With parts from the iGEM registry, our team created a glucose-controlled promoter linked to a yellow fluorescence production gene in E. coli. The concentrations of glucose to which the promoter responds can be determined. Once the concentration is known, the promoter can be mutated so that it will be activated by varying concentrations of glucose and be used as a glucose sensor for people with diabetes. In the future, an insulin gene could be added to this system for use in insulin pumps, where specific glucose levels trigger insulin production in E. coli.</p><br />
<br /><br />
<h3>Our System Overview</h3><br />
<p>We submitted two biobricks to the registry: an intermediate part (http://partsregistry.org/wiki/index.php?title=Part:BBa_K621000) with a ribosome binding site and eYFP, and the intermediate plus the ompC operator (http://partsregistry.org/wiki/index.php?title=Part:BBa_K621001). The ompC operator has three binding sites for phosphorylated OmpR. OmpR and EnvZ work together in a two-component regulatory system as shown in the diagram below. </p><br />
<br /><br />
<p>EnvZ is an inner membrane protein that senses osmolarity. Phosphorlyation of OmpR by EnvZ positively correlates with the osmolarity of the system. Phosphorylated OmpR (OmpR-P) can bind to the three sites on the ompC operator. When one or two of the binding sites are occupied by OmpR-P, RNA polymerase is recruited to begin downstream transcription of the reporter gene, eYFP. However, when all three OmpR binding sites are occupied by OmpR-P, RNA polymerase cannot bind, the reporter gene can no longer be produced, and therefore the system is inhibited. In summary, as osmolarity increases from very low levels, the fluorescence produced by the system also increases until the system reaches a threshold osmolarity. Once the system reaches the threshold, the fluorescence will decrease with increasing osmolarity due to the inherent down-regulation of the system. The activity of the system can be quantified using the fluorescence of the cells because the two-component regulatory system of EnvZ and OmpR regulates transcription of the eYFP gene, dictating the level of fluorescence.</p><br />
<div style="height: 795px"><br />
<h3>Making Our Parts</h3><br />
[[File:Igemmodel3.jpg|right|400px]]<br />
<p>We started with three parts from the iGEM registry:<br />
*BBa_B0032: RBS-3, a medium-strength ribosome binding site<br />
*BBa_E0030: eYFP gene, codes for yellow fluorescence<br />
*BBa_R0082: omp-c operon, contains three binding sites for phosphorylated OmpR</p><br />
</div><br />
<br /><br />
<h3>Testing</h3><br />
<p>To measure fluorescence our team used a 96-well plate reader. Overnight cultures of bacteria were grown in 0.5X LB media and then aliquoted into 1.5ml tubes. Serial dilutions of the desired glucose concentrations were made. Glucose was diluted using 0.5X LB media so that the concentration of LB remained constant. The glucose was administered to the cells in a fashion that did not dilute the cell densities in the cultures. Samples of each treatment were added to a 96-well plate and were imaged using the plate reader. See [[Team:Missouri_Miners/data| data]] page for results. </p><br />
<br /><br />
<h3>Next Steps</h3><br />
<p>In the future, our system has applications in glucose sensing devices. For our system to be sensitive to different changes in osmolarity the OmpC operon will need to be mutated. The mutagenesis process begins with transforming our system into a mutagenic strain of E. coli, in which, the cells have no proof reading capability during DNA replication. In these cells the rate of mutation is increased by 1000x. Using this process mutant strains could be produced, sequenced, and characterized. Possible applications of a characterized system include economical glucose testing strips, a wider range of test strip availability, and precursors to advanced insulin pump systems. </p><br />
</div><br />
</div></div>Eksch9http://2011.igem.org/Team:Missouri_Miners/ProjectTeam:Missouri Miners/Project2011-09-29T02:43:07Z<p>Eksch9: </p>
<hr />
<div>{{OrganizationS&T}}<br />
<br />
<div id="box" style="position: relative; width: 740px; margin-left: 30px; top:-10px; padding: 5px; background-color: #000000;"> <br />
<div id="template" style="font-weight: regular; font-size: medium; color: silver; padding: 5px;"><br />
<h1>Project </h1><br /><br />
<br />
<h3>Abstract</h3><br />
<p>In the bodies of people with diabetes, the ability to recognize and respond to glucose concentrations in the blood has been compromised. As a result, glucose accumulates to dangerous levels. High blood glucose concentrations can cause irreversible damage to critical organs, impairing their functionality. With parts from the iGEM registry, our team created a glucose-controlled promoter linked to a yellow fluorescence production gene in E. coli. The concentrations of glucose to which the promoter responds can be determined. Once the concentration is known, the promoter can be mutated so that it will be activated by varying concentrations of glucose and be used as a glucose sensor for people with diabetes. In the future, an insulin gene could be added to this system for use in insulin pumps, where specific glucose levels trigger insulin production in E. coli.</p><br />
<br /><br />
<h3>Our System Overview</h3><br />
<p>We submitted two biobricks to the registry: an intermediate part (http://partsregistry.org/wiki/index.php?title=Part:BBa_K621000) with a ribosome binding site and eYFP, and the intermediate plus the ompC operator (http://partsregistry.org/wiki/index.php?title=Part:BBa_K621001). The ompC operator has three binding sites for phosphorylated OmpR. OmpR and EnvZ work together in a two-component regulatory system as shown in the diagram below. </p><br />
<br /><br />
<p>EnvZ is an inner membrane protein that senses osmolarity. Phosphorlyation of OmpR by EnvZ positively correlates with the osmolarity of the system. Phosphorylated OmpR (OmpR-P) can bind to the three sites on the ompC operator. When one or two of the binding sites are occupied by OmpR-P, RNA polymerase is recruited to begin downstream transcription of the reporter gene, eYFP. However, when all three OmpR binding sites are occupied by OmpR-P, RNA polymerase cannot bind, the reporter gene can no longer be produced, and therefore the system is inhibited. In summary, as osmolarity increases from very low levels, the fluorescence produced by the system also increases until the system reaches a threshold osmolarity. Once the system reaches the threshold, the fluorescence will decrease with increasing osmolarity due to the inherent down-regulation of the system. The activity of the system can be quantified using the fluorescence of the cells because the two-component regulatory system of EnvZ and OmpR regulates transcription of the eYFP gene, dictating the level of fluorescence.</p><br />
<div style="height: 795px"><br />
<h3>Making Our Parts</h3><br />
[[File:Igemmodel3.jpg|right|400px]]<br />
<p>We started with three parts from the iGEM registry:<br />
*BBa_B0032: RBS-3, a medium-strength ribosome binding site<br />
*BBa_E0030: eYFP gene, codes for yellow fluorescence<br />
*BBa_R0082: omp-c operon, contains three binding sites for phosphorylated OmpR</p><br />
</div><br />
<br /><br />
<h3>Testing</h3><br />
<p>To measure fluorescence our team used a 96-well plate reader. Overnight cultures of bacteria were grown in 0.5X LB media and then aliquoted into 1.5ml tubes. Serial dilutions of the desired glucose concentrations were made. Glucose was diluted using 0.5X LB media so that the concentration of LB remained constant. The glucose was administered to the cells in a fashion that did not dilute the cell densities in the cultures. Samples of each treatment were added to a 96-well plate and were imaged using the plate reader. See [[Team:Missouri_Miners/data| data]] page for results. </p><br />
<br /><br />
<h3>Next Steps</h3><br />
<p>Mutagenesis.</p><br />
</div><br />
</div></div>Eksch9http://2011.igem.org/File:Grown_in_LB.jpgFile:Grown in LB.jpg2011-09-29T02:29:51Z<p>Eksch9: </p>
<hr />
<div></div>Eksch9http://2011.igem.org/Team:Missouri_Miners/DataTeam:Missouri Miners/Data2011-09-29T02:29:26Z<p>Eksch9: </p>
<hr />
<div>{{OrganizationS&T}}<br />
<br />
<html><br />
<div style="width: 740px; margin: 30px; padding: 10px; background-color: #000000; color: silver; font-size: medium"><br />
<h1>Data</h1><br />
<br />
<br />
<br />
<p /> Our part consists of an OmpR binding region that regulates our reporter gene, eyfp. The OmpR binding region is the ompC operator, which has three OmpR binding sites. Our part is regulated by the EnvZ-OmpR two-component regulatory system. EnzZ is aosmolarity sensor on the inner membrane and phosphorylates OmpR in the presence of high osmolarity. When phosphorylated, OmpR binds to the ompC operator and recruits RNAP to transcribe the downstream reporter gene. Fluorescence intensity due to the production of eYfp is a measure of the system's activity. When all three binding sites are occupied by phosphorylated OmpR proteins, RNAP cannot bind to the DNA and transcribe downstream. This part can be used as an osmolarity indicator. Our team used this part as a glucose sensor because the presence of glucose changes osmolarity. Different concentrations of glucose produce different, quantifiable fluorescence intensities.<br/><br/><br />
<br />
<br />
<br />
====Sequence Verification====<br/><br />
This part has been sequence verified and works as expected. <br/><br/><br />
<br />
<br />
====Fluorescence wavelengths====<br/><br />
*'''Excitation max''' - 485nm<br/><br />
*'''Emission max''' - 520nm<br/><br/><br />
<br />
<br />
<br />
====Growth Conditions====<br/><br />
*'''Growth Temperature and Rate''' - E. colistrain DH5alphaused as Chassis, standard growth conditions apply <br/><br />
*'''Growth Media''' - Nutrient Broth or diluted LB media, since OmpR is an osmolarity receptor the cells must be grown in a low osmolarity media to control for compounding factors. To express Fluorescence it is recommended that cultures are grown in liquid media. Colonies growing on plates cannot detect osmolarity and will not activate EnvZ. <br/><br />
<br />
If cells are grown in a high osmolarity media, such as LB broth, the background fluorescence will be higher than an induced glucose response. This is because the system is already primed from the high osmolarity of the media. When additional osmolites are added, the system will down regulate eyfp due to the presence of too many phosphorylated OmpR proteins. Below is a dose response in the presence of LB media. Note how glucose, in these conditions, has an inhibitory effect on the system. <br/><br />
</html><br />
[[Image:Grown in LB.jpg]]<br />
<html><br />
<br/><br/><br />
<br />
====Dose Response====<br/><br />
To control for compounding variables, the cells assayed were grown in 0.5X LB media. This reduced the background osmolarity and allowed our team to measure a valid response. In the graph you can see that the system fluoresces over background at 1% Glucose and then drops back to background at 5% Glucose. The increase at 1% is due to positive regulation by the EnvZ-OmpR system. 5% glucose is a high enough concentration to induce downregulation of the system. Too many phosphorylated OmpR proteins bind the ompC promoter. When all three binding sites are filled RNAP cannot transcribe downstream and fluorescence decreases. <br/><br/><br />
<br />
<br />
<img src="https://static.igem.org/mediawiki/2011/4/48/Dose_response_1.jpg" alt="Dose Response"> <br/><br/><br />
<br />
====Time Trial====<br/><br />
The graph below shows time trials of the system. These time trials were run in the presence 0%, 1%, and 8% glucose. We chose to do this so we could gather more comprehensive data. The first conclusion we can draw from this graph is that the response is fairly consistent over time (there is no peak in fluorescence associated with a specific time point). Also, for each time point between 0.5-4.5 hrs, the curve of the dose response remains consistent. This graph also shows that the assay is not accurate after 4.5 hours. The data beyond 4.5 hours becomes random and inconsistent.<br/><br/><br />
<br />
<br />
<img src="https://static.igem.org/mediawiki/2011/f/fe/Time_Trials_2.jpg" alt="Time Trials"><br/><br/><br />
</div><br />
</html></div>Eksch9http://2011.igem.org/File:Time_Trials_2.jpgFile:Time Trials 2.jpg2011-09-29T01:57:20Z<p>Eksch9: </p>
<hr />
<div></div>Eksch9http://2011.igem.org/Team:Missouri_Miners/DataTeam:Missouri Miners/Data2011-09-29T01:57:03Z<p>Eksch9: </p>
<hr />
<div>{{OrganizationS&T}}<br />
<br />
<html><br />
<div style="width: 740px; margin: 30px; padding: 10px; background-color: #000000; color: silver; font-size: medium"><br />
<h1>Data</h1><br />
<br />
<br />
<br />
<p /> Our part consists of an OmpR binding region that regulates our reporter gene, eyfp. The OmpR binding region is the ompC operator, which has three OmpR binding sites. Our part is regulated by the EnvZ-OmpR two-component regulatory system. EnzZ is aosmolarity sensor on the inner membrane and phosphorylates OmpR in the presence of high osmolarity. When phosphorylated, OmpR binds to the ompC operator and recruits RNAP to transcribe the downstream reporter gene. Fluorescence intensity due to the production of eYfp is a measure of the system's activity. When all three binding sites are occupied by phosphorylated OmpR proteins, RNAP cannot bind to the DNA and transcribe downstream. This part can be used as an osmolarity indicator. Our team used this part as a glucose sensor because the presence of glucose changes osmolarity. Different concentrations of glucose produce different, quantifiable fluorescence intensities. <br />
<br />
<br />
<br />
====Sequence Verification====<br />
This part has been sequence verified and works as expected. <br />
<br />
<br />
====Fluorescence wavelengths====<br />
*'''Excitation max''' - 485nm<br />
*'''Emission max''' - 520nm<br />
<br />
<br />
<br />
====Growth Conditions====<br />
*'''Growth Temperature and Rate''' - E. colistrain DH5alphaused as Chassis, standard growth conditions apply <br />
*'''Growth Media''' - Nutrient Broth or diluted LB media, since OmpR is an osmolarity receptor the cells must be grown in a low osmolarity media to control for compounding factors. . <br />
<br />
If cells are grown in a high osmolarity media, such as LB broth, the background fluorescence will be higher than an induced glucose response. This is because the system is already primed from the high osmolarity of the media. When additional osmolites are added, the system will down regulate eyfp due to the presence of too many phosphorylated OmpR proteins. Below is a dose response in the presence of LB media. Note how glucose, in these conditions, has an inhibitory effect on the system. <br />
<br />
[[Image:Grown in LB.jpg]]<br />
<br />
<br />
====Dose Response====<br />
To control for compounding variables, the cells assayed were grown in 0.5X LB media. This reduced the background osmolarity and allowed our team to measure a valid response. In the graph you can see that the system fluoresces over background at 1% Glucose and then drops back to background at 5% Glucose. The increase at 1% is due to positive regulation by the EnvZ-OmpR system. 5% glucose is a high enough concentration to induce downregulation of the system. Too many phosphorylated OmpR proteins bind the ompC promoter. When all three binding sites are filled RNAP cannot transcribe downstream and fluorescence decreases. <br />
<br />
<br />
===[[Image:Dose_response_1.jpg]]===<br />
<br />
====Time Trial====<br />
The graph below shows time trials of the system. These time trials were run in the presence 0%, 1%, and 8% glucose. We chose to do this so we could gather more comprehensive data. The first conclusion we can draw from this graph is that the response is fairly consistent over time (there is no peak in fluorescence associated with a specific time point). Also, for each time point between 0.5-4.5 hrs, the curve of the dose response remains consistent. This graph also shows that the assay is not accurate after 4.5 hours. The data beyond 4.5 hours becomes random and inconsistent.<br />
<br />
<br />
[[Image:Time_Trials_2.jpg]]</div>Eksch9http://2011.igem.org/Team:Missouri_Miners/PapersTeam:Missouri Miners/Papers2011-09-29T01:15:47Z<p>Eksch9: </p>
<hr />
<div>{{OrganizationS&T}}<br />
<br />
<br />
<div id="box" style="position: relative; width: 740px; margin-left: 30px; top:-20px; padding: 5px; background-color: #000000;"> <br />
<div id="template" style="font-weight: regular; font-size: medium; color: silver; padding: 5px;"><br />
<h1>Research Papers </h1><br /><br />
<br />
<br />
Phosphorylation stimulates the cooperative DNA-binding<br />
properties of the transcription factor OmpR<br />
http://www.pnas.org/content/94/7/2828.full.pdf<br />
<br />
Suppressor Mutations in rpoA Suggest that OmpR Controls<br />
Transcription by Direct Interaction with the<br />
a Subunit of RNA Polymerase<br />
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC212516/<br />
<br />
EnvZ-OmpR Interaction and Osmoregulation in Escherichia coli*<br />
http://www.ncbi.nlm.nih.gov/pubmed/11973328<br />
<br />
Phosphorylation of OmpR by the osmosensor EnvZ modulates<br />
expression of the ompF and ompC genes in Escherichia coli<br />
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC297773/<br />
<br />
Molecular Analysis of the Signaling Pathway between<br />
EnvZ and OmpR in Escherichia coli<br />
http://www.ncbi.nlm.nih.gov/pubmed/1311295<br />
<br />
TWO-COMPONENT SIGNAL TRANSDUCTION<br />
http://www.ncbi.nlm.nih.gov/pubmed/10966457</div>Eksch9http://2011.igem.org/Team:Missouri_Miners/DataTeam:Missouri Miners/Data2011-09-29T01:05:20Z<p>Eksch9: </p>
<hr />
<div>{{OrganizationS&T}}<br />
<br />
<html><br />
<div style="width: 740px; margin: 30px; padding: 10px; background-color: #000000; color: silver; font-size: medium"><br />
<h1>Data</h1><br />
<br />
<br />
<br />
<p /> Our part consists of an OMP-R binding region that regulates our reporter gene, eYFP. The OMP-R binding region is an omp-c operator, which has three OMP-R binding sites. Our part is regulated by the ENVZ-OMPR two-component regulatory system. ENVZ is a osmolarity sensor on the inner membrane and phosphorylates OMP-R in the presence of high osmolarity. When phosphorylated, OMP-R binds to the omp-c operator and recruits RNAP to transcribe the downstream reporter gene. Fluorescence intensity due to the production of eYFP is a measure of the system's activity. When all three binding sites are occupied by phosphorylated OMP-R proteins, RNAP cannot bind to the DNA and transcribe downstream. This part can be used as an osmolarity indicator. Our team used this part as a glucose sensor because the presence of glucose changes osmolarity. Different concentrations of glucose produce different, quantifiable fluorescence intensities. <br />
<br />
<br />
====Fluorescence wavelengths====<br />
*'''Excitation max''' - 485nm<br />
*'''Emission max''' - 520nm<br />
<br />
<br />
<br />
====Growth Conditions====<br />
*'''Growth Temperature and Rate''' - DH5alpha ''E.coli'' used as Chassis, standard growth conditions apply<br />
*'''Growth Media''' - Nutrient Broth, since OMP-R is an osmolarity receptor the cells must be grown in a low osmolarity media (nutrient broth) to control for confounding factors. <br />
<br />
If cells are grown in a high osmolarity media, such as LB broth, the background fluorescence will be higher than an induced glucose response. This is because the system is already primed from the high osmolarity of the media. When additional sources of osmolarity are added, the system will down regulate eYFP due the the presence of too many OMP-R proteins. Below is a dose response in the presence of LB media. Note how glucose, in these conditions, has an inhibitory effect on the system. <br />
<br />
[[Image:Grown in LB.jpg]]<br />
<br />
<br />
====Dose Response====<br />
This part has been sequence verified and works as expected. To control for confounding variables, the cells assayed were grown in 0.5X LB media. This reduced the background osmolarity and allowed our team to measure a valid response. In the graph you can see that the system fluoresces over background at 1% Glucose and then drops back to background at 5% Glucose. The increase at 1% is due to positive regulation of the ENVZ-OMPR system. 5% glucose is a high enough concentration to induce downregulation of the system. Too many OMP-R proteins become phosphorylated and bind the omp-c promoter. When all three binding sites are filled RNAP cannot transcribe downstream and fluorescence decreases. <br />
<br />
<br />
===[[Image:Dose_response_1.jpg]]===<br />
<br />
====Time Trial====<br />
The graph below shows time trials of the system. These time trials were run in the presence 0%, 1%, and 8% glucose. We chose to do this so we could gather more comprehensive data. The first conclusion we can draw from this graph is that the response is fairly consistent over time (there is no peak in fluorescence associated with a specific time point). Also, for each time point between 0.5-4.5 hrs, the curve of the dose response remains consistent. This graph also shows that the assay is not accurate after 4.5 hours. The data beyond 4.5 hours becomes random and inconsistent. <br />
<br />
<br />
[[Image:Time_Trials_1.jpg]]</div>Eksch9http://2011.igem.org/Team:Missouri_Miners/ProjectTeam:Missouri Miners/Project2011-09-29T00:00:47Z<p>Eksch9: </p>
<hr />
<div>{{OrganizationS&T}}<br />
<br />
<br />
<br />
<div id="box" style="position: relative; width: 740px; margin-left: 30px; top:-10px; padding: 5px; background-color: #000000;"> <br />
<div id="template" style="font-weight: regular; font-size: medium; color: silver; padding: 5px;"><br />
<h1>Project </h1><br /><br />
<br />
<br />
Part 1:<br />
<br />
As a standard of iGEM, E.coli will be used as the bacterium for this research. We will incorporate a eYFP gene, ribosome binding site, and Omp-R promoter into E. coli as a plasmid. The plasmid will consist of two main parts, the promoter and a reporter system. The promoter is an osmolarity activated promoter, we dictate this part as OmpR. The reporter system is a DNA sequence which codes for a yellow florescence protein (eYFP) gene with a ribosome binding site (RBS). These parts were prepped to be put together by digesting out the eYFP gene at the Xba1 and Pst1 restriction enzyme sites and opening up the RBS plasmid by cutting at the Spe1 and Pst1 restriction enzyme sites. Gel electrophoresis was used to purify the digested segments and then ligated together creating our intermediate part, BBa_K621000, the reporter system. This DNA was transformed into chemically competent E. coli to amplify our part. After a plasmid prep of our transformation another digest was used to cut out the reporter system, RBS/eYFP, and open up the OmpR plasmid. Another set of gel electrophoresis, purification, transformation, and plasmid prep our part, BBa_K621001 was ready to be tested.<br />
<br />
Part 2:<br />
<br />
Once the three parts are together and in the correct order OmpR/RBS/eYFP testing needed to be done to see if they were in fact in the correct order, and also what concentration of glucose it would fluoresce. For testing to see if our part, BBa_K621001, is in the order its supposed to be the DNA was sequenced. The sequencing data gave us the order of the nucleic acid bases, then the data can be compared with the known sequence of each part. To tell if the part works glucose was added to agar plates at a 25% concentration, no florescence was seen and very little growth occurred. Next we used agar/glucose plates 0%, 1%, 5%, 10%, and 15% concentrations were tried, aging resulting in no florescence. Later it was realized our mistake of trying to use solid media plates for testing a osmolarity sensor instead of liquid media. Osmolarity is the measure of solutes in a solution that change the osmotic pressure of the solution. Making the switch to liquid cultures so the part could sense osmolarity was just what was needed to happen so that our part fluoresced. To get an idea of what concentrations of glucose would still let the part glow 0%, 1%, 3%, 5%, 8%, 10%, 15%, 20,and 25% glucose - LB broth was used. This was later changed from a regular concentration of LB broth to half the normal concentration, because after some research it was seen that glucose and LB broth have very similar osmolarities and so adding one to the other did not cause a change in osmolarity. This problem was quickly fixed by going to .5X concentration LB broth.<br />
<br />
Part 3:<br />
<br />
The next step of the project would be to go through a mutagenesis process to try to change the sensitivity of the OmpR promoter to be able to be turned on by a smaller change in osmolarity. Mutagenesis uses a strain of E. coli, MutD, that has no proof reading capability in its genome. This increases the rate at which mutations occur by a factor of 1000. We</div>Eksch9http://2011.igem.org/File:Dose_response_1.jpgFile:Dose response 1.jpg2011-09-28T23:51:31Z<p>Eksch9: uploaded a new version of &quot;File:Dose response 1.jpg&quot;: Reverted to version as of 23:49, 28 September 2011</p>
<hr />
<div></div>Eksch9http://2011.igem.org/Team:Missouri_Miners/DataTeam:Missouri Miners/Data2011-09-28T23:50:44Z<p>Eksch9: </p>
<hr />
<div>{{OrganizationS&T}}<br />
<br />
<html><br />
<div style="width: 740px; margin: 30px; padding: 10px; background-color: #000000; color: silver; font-size: medium"><br />
<h1>Data</h1><br />
<br />
<br />
<br />
<p /> Our part consists of an OMP-R binding region that regulates our reporter gene, eYFP. The OMP-R binding region is an omp-c operator, which has three OMP-R binding sites. Our part is regulated by the ENVZ-OMPR two-component regulatory system. ENVZ is a osmolarity sensor on the inner membrane and phosphorylates OMP-R in the presence of high osmolarity. When phosphorylated, OMP-R binds to the omp-c operator and recruits RNAP to transcribe the downstream reporter gene. Fluorescence intensity due to the production of eYFP is a measure of the system's activity. When all three binding sites are occupied by phosphorylated OMP-R proteins, RNAP cannot bind to the DNA and transcribe downstream. This part can be used as an osmolarity indicator. Our team used this part as a glucose sensor because the presence of glucose changes osmolarity. Different concentrations of glucose produce different, quantifiable fluorescence intensities. <p /><br />
<br />
<br />
This part has been sequence verified and works as expected. In the first graph you can see that the system fluoresces over background at 1% Glucose and then drops back to background at 5% Glucose. The increase at 1% is due to positive regulation of the ENVZ-OMPR system. 5% glucose is a high enough concentration to induce downregulation of the system. Too many OMP-R proteins become phosphorylated and bind the omp-c promoter. When all three binding sites are filled RNAP cannot transcribe downstream and fluorescence decreases. <br />
<br />
{[Image:Dose_response_1.jpg]] <br />
<br />
The graph below shows time trials of the system. These time trials were run in the presence 0%, 1%, and 8% glucose. We chose to do this so we could gather more comprehensive data. The first conclusion we can draw from this graph is that the response is fairly consistent over time (there is no peak in fluorescence associated with a specific time point). Also, for each time point between 0.5-4.5 hrs, the curve of the dose response remains consistent. This graph also shows that the assay is not accurate after 4.5 hours. The data beyond 4.5 hours becomes random and inconsistent. <br />
<br />
<br />
<br />
<br />
[[Image:Time_Trials_1.jpg]]<br />
<br />
</div><br />
</html></div>Eksch9http://2011.igem.org/File:Time_Trials_1.jpgFile:Time Trials 1.jpg2011-09-28T23:50:04Z<p>Eksch9: </p>
<hr />
<div></div>Eksch9http://2011.igem.org/File:Dose_response_1.jpgFile:Dose response 1.jpg2011-09-28T23:49:32Z<p>Eksch9: uploaded a new version of &quot;File:Dose response 1.jpg&quot;</p>
<hr />
<div></div>Eksch9http://2011.igem.org/Team:Missouri_Miners/DataTeam:Missouri Miners/Data2011-09-28T23:49:17Z<p>Eksch9: </p>
<hr />
<div>{{OrganizationS&T}}<br />
<br />
<html><br />
<div style="width: 740px; margin: 30px; padding: 10px; background-color: #000000; color: silver; font-size: medium"><br />
<h1>Data</h1><br />
<br />
<br />
<br />
<p /> Our part consists of an OMP-R binding region that regulates our reporter gene, eYFP. The OMP-R binding region is an omp-c operator, which has three OMP-R binding sites. Our part is regulated by the ENVZ-OMPR two-component regulatory system. ENVZ is a osmolarity sensor on the inner membrane and phosphorylates OMP-R in the presence of high osmolarity. When phosphorylated, OMP-R binds to the omp-c operator and recruits RNAP to transcribe the downstream reporter gene. Fluorescence intensity due to the production of eYFP is a measure of the system's activity. When all three binding sites are occupied by phosphorylated OMP-R proteins, RNAP cannot bind to the DNA and transcribe downstream. This part can be used as an osmolarity indicator. Our team used this part as a glucose sensor because the presence of glucose changes osmolarity. Different concentrations of glucose produce different, quantifiable fluorescence intensities. <p /><br />
<br />
<br />
This part has been sequence verified and works as expected. In the first graph you can see that the system fluoresces over background at 1% Glucose and then drops back to background at 5% Glucose. The increase at 1% is due to positive regulation of the ENVZ-OMPR system. 5% glucose is a high enough concentration to induce downregulation of the system. Too many OMP-R proteins become phosphorylated and bind the omp-c promoter. When all three binding sites are filled RNAP cannot transcribe downstream and fluorescence decreases. <br />
<br />
[Image:Dose_response_1.jpg] <br />
<br />
The graph below shows time trials of the system. These time trials were run in the presence 0%, 1%, and 8% glucose. We chose to do this so we could gather more comprehensive data. The first conclusion we can draw from this graph is that the response is fairly consistent over time (there is no peak in fluorescence associated with a specific time point). Also, for each time point between 0.5-4.5 hrs, the curve of the dose response remains consistent. This graph also shows that the assay is not accurate after 4.5 hours. The data beyond 4.5 hours becomes random and inconsistent. <br />
<br />
<br />
<br />
<br />
[[Image:Time_Trials_1.jpg]]<br />
<br />
</div><br />
</html></div>Eksch9http://2011.igem.org/File:Dose_response_1.jpgFile:Dose response 1.jpg2011-09-28T23:49:06Z<p>Eksch9: uploaded a new version of &quot;File:Dose response 1.jpg&quot;</p>
<hr />
<div></div>Eksch9http://2011.igem.org/File:Dose_response_1.jpgFile:Dose response 1.jpg2011-09-28T21:37:42Z<p>Eksch9: </p>
<hr />
<div></div>Eksch9http://2011.igem.org/Template:OrganizationS%26TTemplate:OrganizationS&T2011-09-28T11:42:19Z<p>Eksch9: </p>
<hr />
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*<br />
* TERMS OF USE - jQuery Easing<br />
* <br />
* Open source under the BSD License. <br />
* <br />
* Copyright c 2008 George McGinley Smith<br />
* All rights reserved.<br />
* <br />
* Redistribution and use in source and binary forms, with or without modification, <br />
* are permitted provided that the following conditions are met:<br />
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* Redistributions of source code must retain the above copyright notice, this list of <br />
* conditions and the following disclaimer.<br />
* Redistributions in binary form must reproduce the above copyright notice, this list <br />
* of conditions and the following disclaimer in the documentation and/or other materials <br />
* provided with the distribution.<br />
* <br />
* Neither the name of the author nor the names of contributors may be used to endorse <br />
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* <br />
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* COPYRIGHT OWNER OR CONTRIBUTORS BE LIABLE FOR ANY DIRECT, INDIRECT, INCIDENTAL, SPECIAL,<br />
* EXEMPLARY, OR CONSEQUENTIAL DAMAGES (INCLUDING, BUT NOT LIMITED TO, PROCUREMENT OF SUBSTITUTE<br />
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* OF THE POSSIBILITY OF SUCH DAMAGE. <br />
*<br />
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/*<br />
*<br />
* TERMS OF USE - EASING EQUATIONS<br />
* <br />
* Open source under the BSD License. <br />
* <br />
* Copyright c 2001 Robert Penner<br />
* All rights reserved.<br />
* <br />
* Redistribution and use in source and binary forms, with or without modification, <br />
* are permitted provided that the following conditions are met:<br />
* <br />
* Redistributions of source code must retain the above copyright notice, this list of <br />
* conditions and the following disclaimer.<br />
* Redistributions in binary form must reproduce the above copyright notice, this list <br />
* of conditions and the following disclaimer in the documentation and/or other materials <br />
* provided with the distribution.<br />
* <br />
* Neither the name of the author nor the names of contributors may be used to endorse <br />
* or promote products derived from this software without specific prior written permission.<br />
* <br />
* THIS SOFTWARE IS PROVIDED BY THE COPYRIGHT HOLDERS AND CONTRIBUTORS "AS IS" AND ANY <br />
* EXPRESS OR IMPLIED WARRANTIES, INCLUDING, BUT NOT LIMITED TO, THE IMPLIED WARRANTIES OF<br />
* MERCHANTABILITY AND FITNESS FOR A PARTICULAR PURPOSE ARE DISCLAIMED. IN NO EVENT SHALL THE<br />
* COPYRIGHT OWNER OR CONTRIBUTORS BE LIABLE FOR ANY DIRECT, INDIRECT, INCIDENTAL, SPECIAL,<br />
* EXEMPLARY, OR CONSEQUENTIAL DAMAGES (INCLUDING, BUT NOT LIMITED TO, PROCUREMENT OF SUBSTITUTE<br />
* GOODS OR SERVICES; LOSS OF USE, DATA, OR PROFITS; OR BUSINESS INTERRUPTION) HOWEVER CAUSED <br />
* AND ON ANY THEORY OF LIABILITY, WHETHER IN CONTRACT, STRICT LIABILITY, OR TORT (INCLUDING<br />
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* OF THE POSSIBILITY OF SUCH DAMAGE. <br />
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/**<br />
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* @name jquery-lightbox-0.5.css<br />
* @author Leandro Vieira Pinho - http://leandrovieira.com<br />
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* @date April 11, 2008<br />
* @category jQuery plugin<br />
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<a href="https://igem.org/Team.cgi">Profile</a><br />
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<a href="https://2011.igem.org/Team:Missouri_Miners/Project_Background">Project Background</a><br />
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<a href="https://2011.igem.org/Team:Missouri_Miners/Collaboration">Attributions</a> <br />
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<a href="https://2011.igem.org/Team:Missouri_Miners/Organization">Organization</a><br />
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<span class="sdt_active"></span><br />
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<span class="sdt_link">Related Links</span><br />
<span class="sdt_descr"></span><br />
</span><br />
</a><br />
<div class="sdt_box"><br />
<a href="https://2011.igem.org/Team:Missouri_Miners/Papers"> Research Papers</a><br />
<a href="https://2011.igem.org/Team:Missouri_Miners/DiabetesLinks">Diabetes</a><br />
<a href="https://2011.igem.org/Team:Missouri_Miners/GlucoseSensorLinks">Glucose Sensors</a><br />
<a href="https://2011.igem.org/Team:Missouri_Miners/InsulinPumpLinks">Insulin Pumps</a><br />
<a href="https://2011.igem.org/Team:Missouri_Miners/EthicsAndSafetyLinks">Ethics and Safety</a><br />
</div><br />
</li><br />
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</div></div>Eksch9http://2011.igem.org/Team:Missouri_Miners/OrganizationTeam:Missouri Miners/Organization2011-09-28T11:41:38Z<p>Eksch9: Created page with "{{OrganizationS&T}} <div id="box" style="position: relative; width: 740px; margin-left: 30px; top:-1px; padding: 5px; background-color: #000000;"> <div id="template" style="fon..."</p>
<hr />
<div>{{OrganizationS&T}}<br />
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<div id="box" style="position: relative; width: 740px; margin-left: 30px; top:-1px; padding: 5px; background-color: #000000;"> <br />
<div id="template" style="font-weight: regular; font-size: medium; color: silver; padding: 5px;"><br />
<h1>Organization</h1><br /></div>Eksch9http://2011.igem.org/Team:Missouri_Miners/CollaborationTeam:Missouri Miners/Collaboration2011-09-28T11:37:46Z<p>Eksch9: </p>
<hr />
<div>{{OrganizationS&T}}<br />
<br />
<div id="box" style="position: relative; width: 740px; margin-left: 30px; top:-1px; padding: 5px; background-color: #000000;"> <br />
<div id="template" style="font-weight: regular; font-size: medium; color: silver; padding: 5px;"><br />
<h1>Attributions </h1><br /><br />
Our team would like to thank '''Vanessa Kaighin''', of the Missouri S&T cDNA Resource Center, for running our sequencing reactions. Vanessa also helped our team troubleshoot and always provided very helpful advice when we needed it. <br /><br /> We would also like to acknowledge the work of '''Fernando Chial''' from Northwest Missouri State University and '''Christy Kwon''' from Truman State University. They both joined our lab and participated in our research efforts over the summer.<br />
<br /><br /><br />
The team owes a big Thank You to the Missouri S&T '''Department of Biological Sciences'''. The department was kind enough to provide us with equipment and lab space. <br />
<br /><br /><br />
Our iGEM team would also like to thank our advisors, Dr. Dave Westenberg and Dr. Katie Shannon. Our advisors helped the team train new members and provided an endless source of advice for our team. <br />
<br /><br /><br />
Finally, we would like to thank our [[Team:Missouri_Miners/Sponsors|sponsors]] for financing our research and making this all possible.</div>Eksch9http://2011.igem.org/Team:Missouri_Miners/CollaborationTeam:Missouri Miners/Collaboration2011-09-28T11:35:06Z<p>Eksch9: </p>
<hr />
<div>{{OrganizationS&T}}<br />
<br />
<div id="box" style="position: relative; width: 740px; margin-left: 30px; top:-1px; padding: 5px; background-color: #000000;"> <br />
<div id="template" style="font-weight: regular; font-size: medium; color: silver; padding: 5px;"><br />
<h1>Attributions </h1><br /><br />
Our team would like to thank '''Vanessa Kaighin''', of the Missouri S&T cDNA Resource Center, for running our sequencing reactions. Vanessa also helped our team troubleshoot and always provided very helpful advice when we needed it. <br /><br /> We would also like to acknowledge the work of '''Fernando Chial''' from Northwest Missouri State University and '''Christy Kwon''' from Truman State University. They both joined our lab and participated in our research efforts over the summer.<br />
<br /><br /><br />
The team owes a big Thank You to the Missouri S&T '''Department of Biological Sciences'''. The department was kind enough to provide us with equipment and lab space. <br />
<br /><br /><br />
Our iGEM team would also like to thank our advisors, Dr. Dave Westenberg and Dr. Katie Shannon. Our advisors helped the team train new members and provided an endless source of advice for our team. <br />
<br /><br /><br />
Finally, we would like to thank our [[sponsors]] for financing our research and making this all possible.</div>Eksch9http://2011.igem.org/Team:Missouri_Miners/Project_BackgroundTeam:Missouri Miners/Project Background2011-09-28T11:09:55Z<p>Eksch9: </p>
<hr />
<div>{{OrganizationS&T}}<br />
<br />
<div style="width: 740px; margin: 0px 30px; padding: 10px; background-color: #000000; color: silver; font-size: medium"><br />
<h1>Project Background</h1><br />
<br /><br />
<h3>Background</h3><br />
<p>Type one diabetes is characterized by the body’s inability to produce insulin and is caused by the death or malfunction of the ß-cells in the pancreas. Insulin is the hormone that decreases the blood glucose concentration by inducing the cellular intake of glucose.</p><br />
<p>Insulin is produced in the Islets of Langerhans (a section of the pancreas) where glucose in the blood is detected. ß-cells in this region produce insulin by monitoring the ratio of ATP (adenosine triphosphate) versus ADP (adenosine diphosphate). When the ATP level is increased the K+ channels close, creating an increase of Ca+ ions in the cell. The unbalance of ions signals the golgi complex to secrete insulin. If the ß-cells die or malfunction then insulin cannot be produced. (http://www.abcam.com/index.html?pageconfig=resource&rid=10602&pid=7) </p><br />
<br /><br />
<div style="text-align:center"><br />
[[File:Insulin_production_model.jpg]]<br />
</div><br />
<br /><br />
<h3>Project Justification</h3><br />
<p>In the United States alone, it is estimated that 3 million people are affected by type one diabetes. Complications that arise due to this disease include heart disease, stroke, high blood pressure, blindness, kidney disease, and neuropathy. The American Diabetes Association has estimated that diabetes costs America over $200 billion per year in diagnosis and treatment (http://www.diabetes.org ).</p><br />
<br /><br />
<h3>Project Inspiration</h3><br />
<p>Our group was brainstorming project ideas one day when a team member spoke up and announced that he has been afflicted with type one diabetes since he was a child. He asked the group if synthetic biology could be used to help people like him with this disease. Research into the topic soon revealed that, in 2007, the Taipei iGEM team had already come up with an answer (http://parts.mit.edu/igem07/index.php/Taipei). They had designed a system with a glucose sensitive promoter and an eYFP reporter gene. This gene could be substituted for an insulin production gene, so that when glucose is present in the environment, insulin is produced. This system has applications in insulin pump technologies.</p><br />
<p>Unfortunately, biobricks for this complete system were not entered into the registry. Our team decided to continue their work so these biobricks could be available on the registry. In addition to submitting this part to the registry our team wanted to modify the part so that is was sensitive to glucose at multiple concentrations. By doing this, we could use the biobrick to develop an economical glucose sensor.</p><br />
<br /><br />
<br />
[[File:Glucose_Testing.jpg]]<br />
<br />
<h3>Project Goal</h3><br />
<p>Our goal is to model a system that can measure glucose levels at different concentrations. In the future, this system could lead to cheaper blood glucose testing devices or an economic and efficient insulin pump. To achieve this goal we are utilizing the EnvZ-OmpR two-component regulatory system. If the OmpR binding region were to be mutated, theoretically, the system would be sensitive to glucose at different glucose concentrations. Each of these mutations are then be isolated and characterized. Once an adequate number of mutants is identified the fluorescence reporter gene could be substituted with other reporter gene (color gene) to make an inexpensive glucose sensor. eYFP could also be replaced with insulin so the system produces insulin at different concentrations of glucose. In the future, this system could then be used as an insulin pump and replace insulin in the body when needed. </p><br />
</div></div>Eksch9http://2011.igem.org/Template:OrganizationS%26TTemplate:OrganizationS&T2011-09-28T09:29:46Z<p>Eksch9: </p>
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</div></div>Eksch9http://2011.igem.org/Team:Missouri_Miners/Project_BackgroundTeam:Missouri Miners/Project Background2011-09-28T09:28:59Z<p>Eksch9: </p>
<hr />
<div>{{OrganizationS&T}}<br />
<br />
<div style="width: 740px; margin: 0px 30px; padding: 10px; background-color: #000000; color: silver; font-size: medium"><br />
<h1>Project Background</h1><br />
<br /><br />
<h3>Background</h3><br />
<p>Type one diabetes is characterized by the body’s inability to produce insulin and is caused by the death or malfunction of the ß-cells in the pancreas. Insulin is the hormone that decreases the blood glucose concentration by inducing the cellular intake of glucose.</p><br />
<p>Insulin is produced in the Islets of Langerhans (a section of the pancreas) where glucose in the blood is detected. ß-cells in this region produce insulin by monitoring the ratio of ATP (adenosine triphosphate) versus ADP (adenosine diphosphate). When the ATP level is increased the K+ channels close, creating an increase of Ca+ ions in the cell. The unbalance of ions signals the golgi complex to secrete insulin. If the ß-cells die or malfunction then insulin cannot be produced. (http://www.abcam.com/index.html?pageconfig=resource&rid=10602&pid=7) </p><br />
<br /><br />
<div style="text-align:center"><br />
[[File:Insulin_production_model.jpg]]<br />
</div><br />
<br /><br />
<h3>Project Justification</h3><br />
<p>In the United States alone, it is estimated that 3 million people are affected by type one diabetes. Complications that arise due to this disease include heart disease, stroke, high blood pressure, blindness, kidney disease, and neuropathy. The American Diabetes Association has estimated that diabetes costs America over $200 billion per year in diagnosis and treatment (http://www.diabetes.org ).</p><br />
<br /><br />
<h3>Project Inspiration</h3><br />
<p>Our group was brainstorming project ideas one day when a team member spoke up and announced that he has been afflicted with type one diabetes since he was a child. He asked the group if synthetic biology could be used to help people like him with this disease. Research into the topic soon revealed that, in 2007, the Taipei iGEM team had already come up with an answer (http://parts.mit.edu/igem07/index.php/Taipei). They had designed a system with a glucose sensitive promoter and an eYFP reporter gene. This gene could be substituted for an insulin production gene, so that when glucose is present in the environment, insulin is produced. This system has applications in insulin pump technologies.</p><br />
<p>Unfortunately, biobricks for this complete system were not entered into the registry. Our team decided to continue their work so these biobricks could be available on the registry. In addition to submitting this part to the registry our team wanted to modify the part so that is was sensitive to glucose at multiple concentrations. By doing this, we could use the biobrick to develop an economical glucose sensor.</p><br />
<br /><br />
<br />
[[File:Glucose_Testing.jpg]]<br />
<br />
<h3>Project Goal</h3><br />
<p>Our goal is to model a system that can measure glucose levels at different concentrations. In the future, this system could lead to cheaper blood glucose testing devices or a economic and efficient insulin pump. To achieve this goal we are utilizing the EnvZ-OmpR two-component regulatory system. This pathway is endogenous to E.Coli and has been well characterized. EnvZ is a osmolarity sensor on the inner membrane of E.Coli. When glucose is added to the environment it induces a change in osmolarity and activates EnvZ. EnvZ then phosphorylates the OmpR transcription factor. When OmpR is phosphorylated it binds to the OmpR binding region and recruits RNA polymerase to begin downstream transcription. If the OmpR binding region were to be mutated</p><br />
</div></div>Eksch9http://2011.igem.org/Team:Missouri_Miners/Project_BackgroundTeam:Missouri Miners/Project Background2011-09-28T09:28:41Z<p>Eksch9: </p>
<hr />
<div>{{OrganizationS&T}}<br />
<br />
<div style="width: 740px; margin: 0px 30px; padding: 10px; background-color: #000000; color: silver; font-size: medium"><br />
<h1>Glucose Sensors</h1><br />
<br /><br />
<h3>Background</h3><br />
<p>Type one diabetes is characterized by the body’s inability to produce insulin and is caused by the death or malfunction of the ß-cells in the pancreas. Insulin is the hormone that decreases the blood glucose concentration by inducing the cellular intake of glucose.</p><br />
<p>Insulin is produced in the Islets of Langerhans (a section of the pancreas) where glucose in the blood is detected. ß-cells in this region produce insulin by monitoring the ratio of ATP (adenosine triphosphate) versus ADP (adenosine diphosphate). When the ATP level is increased the K+ channels close, creating an increase of Ca+ ions in the cell. The unbalance of ions signals the golgi complex to secrete insulin. If the ß-cells die or malfunction then insulin cannot be produced. (http://www.abcam.com/index.html?pageconfig=resource&rid=10602&pid=7) </p><br />
<br /><br />
<div style="text-align:center"><br />
[[File:Insulin_production_model.jpg]]<br />
</div><br />
<br /><br />
<h3>Project Justification</h3><br />
<p>In the United States alone, it is estimated that 3 million people are affected by type one diabetes. Complications that arise due to this disease include heart disease, stroke, high blood pressure, blindness, kidney disease, and neuropathy. The American Diabetes Association has estimated that diabetes costs America over $200 billion per year in diagnosis and treatment (http://www.diabetes.org ).</p><br />
<br /><br />
<h3>Project Inspiration</h3><br />
<p>Our group was brainstorming project ideas one day when a team member spoke up and announced that he has been afflicted with type one diabetes since he was a child. He asked the group if synthetic biology could be used to help people like him with this disease. Research into the topic soon revealed that, in 2007, the Taipei iGEM team had already come up with an answer (http://parts.mit.edu/igem07/index.php/Taipei). They had designed a system with a glucose sensitive promoter and an eYFP reporter gene. This gene could be substituted for an insulin production gene, so that when glucose is present in the environment, insulin is produced. This system has applications in insulin pump technologies.</p><br />
<p>Unfortunately, biobricks for this complete system were not entered into the registry. Our team decided to continue their work so these biobricks could be available on the registry. In addition to submitting this part to the registry our team wanted to modify the part so that is was sensitive to glucose at multiple concentrations. By doing this, we could use the biobrick to develop an economical glucose sensor.</p><br />
<br /><br />
<br />
[[File:Glucose_Testing.jpg]]<br />
<br />
<h3>Project Goal</h3><br />
<p>Our goal is to model a system that can measure glucose levels at different concentrations. In the future, this system could lead to cheaper blood glucose testing devices or a economic and efficient insulin pump. To achieve this goal we are utilizing the EnvZ-OmpR two-component regulatory system. This pathway is endogenous to E.Coli and has been well characterized. EnvZ is a osmolarity sensor on the inner membrane of E.Coli. When glucose is added to the environment it induces a change in osmolarity and activates EnvZ. EnvZ then phosphorylates the OmpR transcription factor. When OmpR is phosphorylated it binds to the OmpR binding region and recruits RNA polymerase to begin downstream transcription. If the OmpR binding region were to be mutated</p><br />
</div></div>Eksch9http://2011.igem.org/Team:Missouri_Miners/Project_BackgroundTeam:Missouri Miners/Project Background2011-09-28T09:27:57Z<p>Eksch9: </p>
<hr />
<div>{{OrganizationS&T}}<br />
<br />
<div id="box" style="position: relative; width: 740px; margin-left: 30px; top:-1px; padding: 5px; background-color: #000000;"> <br />
<div id="template" style="font-weight: regular; font-size: medium; color: silver; padding: 5px;"><br />
<h1>Project Background </h1><br /></div>Eksch9http://2011.igem.org/Team:Missouri_Miners/Project_BackgroundTeam:Missouri Miners/Project Background2011-09-28T09:27:35Z<p>Eksch9: Created page with "{{OrganizationS&T}} <div id="box" style="position: relative; width: 740px; margin-left: 30px; top:-1px; padding: 5px; background-color: #000000;"> <div id="template" style="fon..."</p>
<hr />
<div>{{OrganizationS&T}}<br />
<br />
<div id="box" style="position: relative; width: 740px; margin-left: 30px; top:-1px; padding: 5px; background-color: #000000;"> <br />
<div id="template" style="font-weight: regular; font-size: medium; color: silver; padding: 5px;"><br />
<h1>Team Page </h1><br /></div>Eksch9http://2011.igem.org/Team:Missouri_Miners/Glucose_SensorTeam:Missouri Miners/Glucose Sensor2011-09-28T09:26:06Z<p>Eksch9: </p>
<hr />
<div>{{OrganizationS&T}}<br />
<br />
<div style="width: 740px; margin: 0px 30px; padding: 10px; background-color: #000000; color: silver; font-size: medium"><br />
<h1>Glucose Sensors</h1><br />
<br /><br />
<h3>Background</h3><br />
<p>Type one diabetes is characterized by the body’s inability to produce insulin and is caused by the death or malfunction of the ß-cells in the pancreas. Insulin is the hormone that decreases the blood glucose concentration by inducing the cellular intake of glucose.</p><br />
<p>Insulin is produced in the Islets of Langerhans (a section of the pancreas) where glucose in the blood is detected. ß-cells in this region produce insulin by monitoring the ratio of ATP (adenosine triphosphate) versus ADP (adenosine diphosphate). When the ATP level is increased the K+ channels close, creating an increase of Ca+ ions in the cell. The unbalance of ions signals the golgi complex to secrete insulin. If the ß-cells die or malfunction then insulin cannot be produced. (http://www.abcam.com/index.html?pageconfig=resource&rid=10602&pid=7) </p><br />
<br /><br />
<div style="text-align:center"><br />
[[File:Insulin_production_model.jpg]]<br />
</div><br />
<br /><br />
<h3>Project Justification</h3><br />
<p>In the United States alone, it is estimated that 3 million people are affected by type one diabetes. Complications that arise due to this disease include heart disease, stroke, high blood pressure, blindness, kidney disease, and neuropathy. The American Diabetes Association has estimated that diabetes costs America over $200 billion per year in diagnosis and treatment (http://www.diabetes.org ).</p><br />
<br /><br />
<h3>Project Inspiration</h3><br />
<p>Our group was brainstorming project ideas one day when a team member spoke up and announced that he has been afflicted with type one diabetes since he was a child. He asked the group if synthetic biology could be used to help people like him with this disease. Research into the topic soon revealed that, in 2007, the Taipei iGEM team had already come up with an answer (https://2007.igem.org/Taipei). They had designed a system with a glucose sensitive promoter and an eYFP reporter gene. This gene could be substituted for an insulin production gene, so that when glucose is present in the environment, insulin is produced. This system has applications in insulin pump technologies.</p><br />
<p>Unfortunately, biobricks for this complete system were not entered into the registry. Our team decided to continue their work so these biobricks could be available on the registry. In addition to submitting this part to the registry our team wanted to modify the part so that is was sensitive to glucose at multiple concentrations. By doing this, we could use the biobrick to develop an economical glucose sensor.</p><br />
<br /><br />
<br />
[[File:Glucose_Testing.jpg]]<br />
<br />
<h3>Project Goal</h3><br />
<p>Our goal is to model a system that can measure glucose levels at different concentrations. In the future, this system could lead to cheaper blood glucose testing devices or a economic and efficient insulin pump. To achieve this goal we are utilizing the EnvZ-OmpR two-component regulatory system. This pathway is endogenous to E.Coli and has been well characterized. EnvZ is a osmolarity sensor on the inner membrane of E.Coli. When glucose is added to the environment it induces a change in osmolarity and activates EnvZ. EnvZ then phosphorylates the OmpR transcription factor. When OmpR is phosphorylated it binds to the OmpR binding region and recruits RNA polymerase to begin downstream transcription. If the OmpR binding region were to be mutated</p><br />
</div></div>Eksch9http://2011.igem.org/Team:Missouri_Miners/Glucose_SensorTeam:Missouri Miners/Glucose Sensor2011-09-28T09:05:59Z<p>Eksch9: </p>
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<h1>Glucose Sensors</h1><br />
<br /><br />
<h3>Background</h3><br />
<p>Type one diabetes is characterized by the body’s inability to produce insulin and is caused by the death or malfunction of the ß-cells in the pancreas. Insulin is the hormone that decreases the blood glucose concentration by inducing the cellular intake of glucose.</p><br />
<p>Insulin is produced in the Islets of Langerhans (a section of the pancreas) where glucose in the blood is detected. ß-cells in this region produce insulin by monitoring the ratio of ATP (adenosine triphosphate) versus ADP (adenosine diphosphate). When the ATP level is increased the K+ channels close, creating an increase of Ca+ ions in the cell. The unbalance of ions signals the golgi complex to secrete insulin. If the ß-cells die or malfunction then insulin cannot be produced. (http://www.abcam.com/index.html?pageconfig=resource&rid=10602&pid=7) </p><br />
<br /><br />
<div style="text-align:center"><br />
[[File:Insulin_production_model.jpg]]<br />
</div><br />
<br /><br />
<h3>Project Justification</h3><br />
<p>In the United States alone, it is estimated that 3 million people are affected by type one diabetes. Complications that arise due to this disease include heart disease, stroke, high blood pressure, blindness, kidney disease, and neuropathy. The American Diabetes Association has estimated that diabetes costs America over $200 billion per year in diagnosis and treatment (http://www.diabetes.org ).</p><br />
<br /><br />
<h3>Project Inspiration</h3><br />
<p>Our group was brainstorming project ideas one day when a team member spoke up and announced that he has been afflicted with type one diabetes since he was a child. He asked the group if synthetic biology could be used to help people like him with this disease. Research into the topic soon revealed that, in 2007, the Taipei iGEM team had already come up with an answer (http://parts.mit.edu/igem07/index.php/Taipei). They had designed a system with a glucose sensitive promoter and an eYFP reporter gene. This gene could be substituted for an insulin production gene, so that when glucose is present in the environment, insulin is produced. This system has applications in insulin pump technologies.</p><br />
<p>Unfortunately, biobricks for this complete system were not entered into the registry. Our team decided to continue their work so these biobricks could be available on the registry. In addition to submitting this part to the registry our team wanted to modify the part so that is was sensitive to glucose at multiple concentrations. By doing this, we could use the biobrick to develop an economical glucose sensor.</p><br />
<br /><br />
<br />
[[File:Glucose_Testing.jpg]]<br />
<br />
<h3>Project Goal</h3><br />
<p>Our goal is to model a system that can measure glucose levels and produce insulin as needed. In the future, this system could lead to cheaper blood glucose testing devices or a economic and efficient insulin pump. To achieve this goal we are utilizing the EnvZ-OmpR signal transduction pathway. This pathway is endogenous to E.Coli and has been well characterized. EnvZ is a osmolarity sensor on the inner membrane of E.Coli. When glucose is added to the environment it induces a change in osmolarity and activates EnvZ. EnvZ then phosphorylates the OmpR transcription factor. When OmpR is phosphorylated it binds to the OmpR promoter and recruits RNA polymerase to begin downstream transcription.</p><br />
</div></div>Eksch9http://2011.igem.org/File:Glucose_Testing.jpgFile:Glucose Testing.jpg2011-09-28T09:04:19Z<p>Eksch9: </p>
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<div></div>Eksch9http://2011.igem.org/Team:Missouri_Miners/EthicsAndSafetyLinksTeam:Missouri Miners/EthicsAndSafetyLinks2011-09-28T08:56:35Z<p>Eksch9: </p>
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<h1>Ethics and Safety </h1><br /><br />
<br />
Missouri S&T Lab Safety<br />
http://ehs.mst.edu/labsafety/biologicalsafety/index.html<br />
<br />
Safety in the Biology Laboratory<br />
http://www.sfponline.org/Uploads/labsafetyandequipment.pdf<br />
<br />
Laboratory Safety Links<br />
http://carnegiescience.edu/first_light_case/horn/labsafety.html<br />
<br />
American Physiological Society: Ethics and scientific publication<br />
http://advan.physiology.org/content/29/2/59.abstract<br />
<br />
National Institute of Environmental Health Services: What is Ethics in Research & Why is It Important?<br />
http://www.niehs.nih.gov/research/resources/bioethics/whatis.cfm</div>Eksch9http://2011.igem.org/Team:Missouri_Miners/InsulinPumpLinksTeam:Missouri Miners/InsulinPumpLinks2011-09-28T08:51:11Z<p>Eksch9: </p>
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<h1>Insulin Pumps </h1><br /><br />
<br />
New England Journal of Medicine: Insulin Pumps<br />
http://www.nejm.org/doi/full/10.1056/NEJMoa1002853<br />
<br />
PubMed: Clinical experience with an integrated continuous glucose sensor/insulin pump platform<br />
http://www.ncbi.nlm.nih.gov/pubmed/17142207<br />
<br />
American Diabetes Association: Fully Automated Closed-Loop Insulin Delivery Versus Semiautomated Hybrid Control in Pediatric Patients With Type 1 Diabetes Using an Artificial Pancreas<br />
http://care.diabetesjournals.org/content/31/5/934.short<br />
<br />
American Diabetes Association: Clinical Trial of Programmable Implantable Insulin Pump For Type I Diabetes<br />
http://care.diabetesjournals.org/content/15/7/877<br />
<br />
Types of Insulin Pumps<br />
http://www.diabetes-insulin-pump-therapy.com/types-of-insulin-pump.html</div>Eksch9http://2011.igem.org/Team:Missouri_Miners/GlucoseSensorLinksTeam:Missouri Miners/GlucoseSensorLinks2011-09-28T08:44:54Z<p>Eksch9: </p>
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<h1>Glucose Sensors</h1><br /><br />
<br />
SPIE Digital Library: Optical glucose sensing in biological fluids<br />
http://spiedigitallibrary.org/jbo/resource/1/jbopfo/v5/i1/p5_s1?isAuthorized=no<br />
<br />
Technology Review: Optical Glucose Sensor<br />
http://www.technologyreview.com/biotech/17943/<br />
<br />
MedGadget:Glowing Glucose Sensor for Long-Term Implanted Monitoring<br />
http://medgadget.com/2011/08/glowing-glucose-sensor-for-long-term-implanted-monitoring.html<br />
<br />
Science Daily: Diabetes Care<br />
http://www.sciencedaily.com/releases/2010/07/100728144347.htm</div>Eksch9http://2011.igem.org/Team:Missouri_Miners/GlucoseSensorLinksTeam:Missouri Miners/GlucoseSensorLinks2011-09-28T08:40:11Z<p>Eksch9: </p>
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<h1>Glucose Sensors</h1><br /><br />
<br />
SPIE Digital Library: Optical glucose sensing in biological fluids<br />
http://spiedigitallibrary.org/jbo/resource/1/jbopfo/v5/i1/p5_s1?isAuthorized=no<br />
<br />
<br />
http://www.technologyreview.com/biotech/17943/<br />
<br />
<br />
http://medgadget.com/2011/08/glowing-glucose-sensor-for-long-term-implanted-monitoring.html<br />
<br />
<br />
http://www.sciencedaily.com/releases/2010/07/100728144347.htm</div>Eksch9http://2011.igem.org/Team:Missouri_Miners/DiabetesLinksTeam:Missouri Miners/DiabetesLinks2011-09-28T08:30:09Z<p>Eksch9: </p>
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<h1> Diabetes </h1><br /><br />
<br />
<br />
American Diabetes Association <br />
http://www.diabetes.org/<br />
<br />
New York Times Health Guide<br />
http://health.nytimes.com/health/guides/disease/diabetes/overview.html<br />
<br />
PubMed Health<br />
http://www.ncbi.nlm.nih.gov/pubmedhealth/PMH0002194/<br />
<br />
WebMD<br />
http://diabetes.webmd.com/diabetes-diet-healthy-diet-basics</div>Eksch9http://2011.igem.org/Team:Missouri_Miners/PartsTeam:Missouri Miners/Parts2011-09-28T05:48:45Z<p>Eksch9: </p>
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<h1>Parts </h1><br /><br />
<br />
<br />
<groupparts>iGEM011 Missouri_Miners</groupparts></div>Eksch9http://2011.igem.org/Team:Missouri_MinersTeam:Missouri Miners2011-09-28T05:48:12Z<p>Eksch9: </p>
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<h1>Welcome to the Missouri Miners Wiki page! </h1><br /><br />
<br />
The Team of 2011<br />
<br /><br />
<html><embed type="application/x-shockwave-flash" src="https://picasaweb.google.com/s/c/bin/slideshow.swf" width="700" height="500" flashvars="host=picasaweb.google.com&hl=en_US&feat=flashalbum&RGB=0x000000&feed=https%3A%2F%2Fpicasaweb.google.com%2Fdata%2Ffeed%2Fapi%2Fuser%2F111761835993314772451%2Falbumid%2F5654969572600636673%3Falt%3Drss%26kind%3Dphoto%26hl%3Den_US" pluginspage="http://www.macromedia.com/go/getflashplayer"></embed><br />
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<br /><br />
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<p>We are the Missouri Miners from Missouri University of Science and Technology. Our team formed in 2007 and we've been working hard developing iGEM projects ever since. In the beginning, the team only had a handful of people but, recently, we have grown to 30 hardworking members.</p> <br />
<br />
<br />
<br /><br />
<p>Our team is a student organization at S&T and we have been working hard to establish ourselves among the other well known student groups. We are also working towards becoming S&T's newest Student Design Team. Being at a science and technology university, Design Teams have a strong tradition on our campus and are highly respected. We hope to bring a new design aspect to our school and promote the usefulness of synthetic biology.</p><br />
<br />
<br /><br />
<p>We are very excited to participate in the iGEM 2011 Jamboree!</p><br />
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<br></div>Eksch9http://2011.igem.org/Team:Missouri_Miners/PartsTeam:Missouri Miners/Parts2011-09-28T05:45:46Z<p>Eksch9: </p>
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<groupparts>iGEM011 Missouri_Miners</groupparts></div>Eksch9http://2011.igem.org/Team:Missouri_Miners/CollaborationTeam:Missouri Miners/Collaboration2011-09-28T02:48:14Z<p>Eksch9: </p>
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<h1>Attributions </h1><br /><br />
Our team would like to thank '''Vanessa Kaighin''', of the Missouri S&T cDNA Resource Center, for running our sequencing reactions. <br /><br /> We would also like to acknowledge the work of '''Fernando Chial''' from Northwest Missouri State University and '''Christy Kwon''' from Truman State University. They both joined our lab and participated in our research efforts over the summer.</div>Eksch9http://2011.igem.org/Team:Missouri_Miners/DiabetesLinksTeam:Missouri Miners/DiabetesLinks2011-09-27T06:57:45Z<p>Eksch9: </p>
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<h1> Diabetes </h1><br /><br />
<br />
<br />
American Diabetes Association [http://www.diabetes.org/]</div>Eksch9http://2011.igem.org/Template:OrganizationS%26TTemplate:OrganizationS&T2011-09-27T06:24:20Z<p>Eksch9: </p>
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* of conditions and the following disclaimer in the documentation and/or other materials <br />
* provided with the distribution.<br />
* <br />
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* <br />
* THIS SOFTWARE IS PROVIDED BY THE COPYRIGHT HOLDERS AND CONTRIBUTORS "AS IS" AND ANY <br />
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* MERCHANTABILITY AND FITNESS FOR A PARTICULAR PURPOSE ARE DISCLAIMED. IN NO EVENT SHALL THE<br />
* COPYRIGHT OWNER OR CONTRIBUTORS BE LIABLE FOR ANY DIRECT, INDIRECT, INCIDENTAL, SPECIAL,<br />
* EXEMPLARY, OR CONSEQUENTIAL DAMAGES (INCLUDING, BUT NOT LIMITED TO, PROCUREMENT OF SUBSTITUTE<br />
* GOODS OR SERVICES; LOSS OF USE, DATA, OR PROFITS; OR BUSINESS INTERRUPTION) HOWEVER CAUSED <br />
* AND ON ANY THEORY OF LIABILITY, WHETHER IN CONTRACT, STRICT LIABILITY, OR TORT (INCLUDING<br />
* NEGLIGENCE OR OTHERWISE) ARISING IN ANY WAY OUT OF THE USE OF THIS SOFTWARE, EVEN IF ADVISED <br />
* OF THE POSSIBILITY OF SUCH DAMAGE. <br />
*<br />
*/<br />
<br />
// t: current time, b: begInnIng value, c: change In value, d: duration<br />
jQuery.easing['jswing'] = jQuery.easing['swing'];<br />
<br />
jQuery.extend( jQuery.easing,<br />
{<br />
def: 'easeOutQuad',<br />
swing: function (x, t, b, c, d) {<br />
//alert(jQuery.easing.default);<br />
return jQuery.easing[jQuery.easing.def](x, t, b, c, d);<br />
},<br />
easeInQuad: function (x, t, b, c, d) {<br />
return c*(t/=d)*t + b;<br />
},<br />
easeOutQuad: function (x, t, b, c, d) {<br />
return -c *(t/=d)*(t-2) + b;<br />
},<br />
easeInOutQuad: function (x, t, b, c, d) {<br />
if ((t/=d/2) < 1) return c/2*t*t + b;<br />
return -c/2 * ((--t)*(t-2) - 1) + b;<br />
},<br />
easeInCubic: function (x, t, b, c, d) {<br />
return c*(t/=d)*t*t + b;<br />
},<br />
easeOutCubic: function (x, t, b, c, d) {<br />
return c*((t=t/d-1)*t*t + 1) + b;<br />
},<br />
easeInOutCubic: function (x, t, b, c, d) {<br />
if ((t/=d/2) < 1) return c/2*t*t*t + b;<br />
return c/2*((t-=2)*t*t + 2) + b;<br />
},<br />
easeInQuart: function (x, t, b, c, d) {<br />
return c*(t/=d)*t*t*t + b;<br />
},<br />
easeOutQuart: function (x, t, b, c, d) {<br />
return -c * ((t=t/d-1)*t*t*t - 1) + b;<br />
},<br />
easeInOutQuart: function (x, t, b, c, d) {<br />
if ((t/=d/2) < 1) return c/2*t*t*t*t + b;<br />
return -c/2 * ((t-=2)*t*t*t - 2) + b;<br />
},<br />
easeInQuint: function (x, t, b, c, d) {<br />
return c*(t/=d)*t*t*t*t + b;<br />
},<br />
easeOutQuint: function (x, t, b, c, d) {<br />
return c*((t=t/d-1)*t*t*t*t + 1) + b;<br />
},<br />
easeInOutQuint: function (x, t, b, c, d) {<br />
if ((t/=d/2) < 1) return c/2*t*t*t*t*t + b;<br />
return c/2*((t-=2)*t*t*t*t + 2) + b;<br />
},<br />
easeInSine: function (x, t, b, c, d) {<br />
return -c * Math.cos(t/d * (Math.PI/2)) + c + b;<br />
},<br />
easeOutSine: function (x, t, b, c, d) {<br />
return c * Math.sin(t/d * (Math.PI/2)) + b;<br />
},<br />
easeInOutSine: function (x, t, b, c, d) {<br />
return -c/2 * (Math.cos(Math.PI*t/d) - 1) + b;<br />
},<br />
easeInExpo: function (x, t, b, c, d) {<br />
return (t==0) ? b : c * Math.pow(2, 10 * (t/d - 1)) + b;<br />
},<br />
easeOutExpo: function (x, t, b, c, d) {<br />
return (t==d) ? b+c : c * (-Math.pow(2, -10 * t/d) + 1) + b;<br />
},<br />
easeInOutExpo: function (x, t, b, c, d) {<br />
if (t==0) return b;<br />
if (t==d) return b+c;<br />
if ((t/=d/2) < 1) return c/2 * Math.pow(2, 10 * (t - 1)) + b;<br />
return c/2 * (-Math.pow(2, -10 * --t) + 2) + b;<br />
},<br />
easeInCirc: function (x, t, b, c, d) {<br />
return -c * (Math.sqrt(1 - (t/=d)*t) - 1) + b;<br />
},<br />
easeOutCirc: function (x, t, b, c, d) {<br />
return c * Math.sqrt(1 - (t=t/d-1)*t) + b;<br />
},<br />
easeInOutCirc: function (x, t, b, c, d) {<br />
if ((t/=d/2) < 1) return -c/2 * (Math.sqrt(1 - t*t) - 1) + b;<br />
return c/2 * (Math.sqrt(1 - (t-=2)*t) + 1) + b;<br />
},<br />
easeInElastic: function (x, t, b, c, d) {<br />
var s=1.70158;var p=0;var a=c;<br />
if (t==0) return b; if ((t/=d)==1) return b+c; if (!p) p=d*.3;<br />
if (a < Math.abs(c)) { a=c; var s=p/4; }<br />
else var s = p/(2*Math.PI) * Math.asin (c/a);<br />
return -(a*Math.pow(2,10*(t-=1)) * Math.sin( (t*d-s)*(2*Math.PI)/p )) + b;<br />
},<br />
easeOutElastic: function (x, t, b, c, d) {<br />
var s=1.70158;var p=0;var a=c;<br />
if (t==0) return b; if ((t/=d)==1) return b+c; if (!p) p=d*.3;<br />
if (a < Math.abs(c)) { a=c; var s=p/4; }<br />
else var s = p/(2*Math.PI) * Math.asin (c/a);<br />
return a*Math.pow(2,-10*t) * Math.sin( (t*d-s)*(2*Math.PI)/p ) + c + b;<br />
},<br />
easeInOutElastic: function (x, t, b, c, d) {<br />
var s=1.70158;var p=0;var a=c;<br />
if (t==0) return b; if ((t/=d/2)==2) return b+c; if (!p) p=d*(.3*1.5);<br />
if (a < Math.abs(c)) { a=c; var s=p/4; }<br />
else var s = p/(2*Math.PI) * Math.asin (c/a);<br />
if (t < 1) return -.5*(a*Math.pow(2,10*(t-=1)) * Math.sin( (t*d-s)*(2*Math.PI)/p )) + b;<br />
return a*Math.pow(2,-10*(t-=1)) * Math.sin( (t*d-s)*(2*Math.PI)/p )*.5 + c + b;<br />
},<br />
easeInBack: function (x, t, b, c, d, s) {<br />
if (s == undefined) s = 1.70158;<br />
return c*(t/=d)*t*((s+1)*t - s) + b;<br />
},<br />
easeOutBack: function (x, t, b, c, d, s) {<br />
if (s == undefined) s = 1.70158;<br />
return c*((t=t/d-1)*t*((s+1)*t + s) + 1) + b;<br />
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if ((t/=d/2) < 1) return c/2*(t*t*(((s*=(1.525))+1)*t - s)) + b;<br />
return c/2*((t-=2)*t*(((s*=(1.525))+1)*t + s) + 2) + b;<br />
},<br />
easeInBounce: function (x, t, b, c, d) {<br />
return c - jQuery.easing.easeOutBounce (x, d-t, 0, c, d) + b;<br />
},<br />
easeOutBounce: function (x, t, b, c, d) {<br />
if ((t/=d) < (1/2.75)) {<br />
return c*(7.5625*t*t) + b;<br />
} else if (t < (2/2.75)) {<br />
return c*(7.5625*(t-=(1.5/2.75))*t + .75) + b;<br />
} else if (t < (2.5/2.75)) {<br />
return c*(7.5625*(t-=(2.25/2.75))*t + .9375) + b;<br />
} else {<br />
return c*(7.5625*(t-=(2.625/2.75))*t + .984375) + b;<br />
}<br />
},<br />
easeInOutBounce: function (x, t, b, c, d) {<br />
if (t < d/2) return jQuery.easing.easeInBounce (x, t*2, 0, c, d) * .5 + b;<br />
return jQuery.easing.easeOutBounce (x, t*2-d, 0, c, d) * .5 + c*.5 + b;<br />
}<br />
});<br />
<br />
/*<br />
*<br />
* TERMS OF USE - EASING EQUATIONS<br />
* <br />
* Open source under the BSD License. <br />
* <br />
* Copyright c 2001 Robert Penner<br />
* All rights reserved.<br />
* <br />
* Redistribution and use in source and binary forms, with or without modification, <br />
* are permitted provided that the following conditions are met:<br />
* <br />
* Redistributions of source code must retain the above copyright notice, this list of <br />
* conditions and the following disclaimer.<br />
* Redistributions in binary form must reproduce the above copyright notice, this list <br />
* of conditions and the following disclaimer in the documentation and/or other materials <br />
* provided with the distribution.<br />
* <br />
* Neither the name of the author nor the names of contributors may be used to endorse <br />
* or promote products derived from this software without specific prior written permission.<br />
* <br />
* THIS SOFTWARE IS PROVIDED BY THE COPYRIGHT HOLDERS AND CONTRIBUTORS "AS IS" AND ANY <br />
* EXPRESS OR IMPLIED WARRANTIES, INCLUDING, BUT NOT LIMITED TO, THE IMPLIED WARRANTIES OF<br />
* MERCHANTABILITY AND FITNESS FOR A PARTICULAR PURPOSE ARE DISCLAIMED. IN NO EVENT SHALL THE<br />
* COPYRIGHT OWNER OR CONTRIBUTORS BE LIABLE FOR ANY DIRECT, INDIRECT, INCIDENTAL, SPECIAL,<br />
* EXEMPLARY, OR CONSEQUENTIAL DAMAGES (INCLUDING, BUT NOT LIMITED TO, PROCUREMENT OF SUBSTITUTE<br />
* GOODS OR SERVICES; LOSS OF USE, DATA, OR PROFITS; OR BUSINESS INTERRUPTION) HOWEVER CAUSED <br />
* AND ON ANY THEORY OF LIABILITY, WHETHER IN CONTRACT, STRICT LIABILITY, OR TORT (INCLUDING<br />
* NEGLIGENCE OR OTHERWISE) ARISING IN ANY WAY OUT OF THE USE OF THIS SOFTWARE, EVEN IF ADVISED <br />
* OF THE POSSIBILITY OF SUCH DAMAGE. <br />
*<br />
*/<br />
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/**<br />
* jQuery lightBox plugin<br />
* This jQuery plugin was inspired and based on Lightbox 2 by Lokesh Dhakar (http://www.huddletogether.com/projects/lightbox2/)<br />
* and adapted to me for use like a plugin from jQuery.<br />
* @name jquery-lightbox-0.5.css<br />
* @author Leandro Vieira Pinho - http://leandrovieira.com<br />
* @version 0.5<br />
* @date April 11, 2008<br />
* @category jQuery plugin<br />
* @copyright (c) 2008 Leandro Vieira Pinho (leandrovieira.com)<br />
* @license CCAttribution-ShareAlike 2.5 Brazil - http://creativecommons.org/licenses/by-sa/2.5/br/deed.en_US<br />
* @example Visit http://leandrovieira.com/projects/jquery/lightbox/ for more informations about this jQuery plugin<br />
*/<br />
#jquery-overlay {<br />
position: absolute;<br />
top: 0;<br />
left: 0;<br />
z-index: 90;<br />
width: 100%;<br />
height: 500px;<br />
}<br />
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}<br />
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#lightbox-container-image-box {<br />
position: relative;<br />
background-color: #fff;<br />
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position: absolute;<br />
top: 40%;<br />
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#lightbox-nav a { outline: none;}<br />
#lightbox-nav-btnPrev, #lightbox-nav-btnNext {<br />
width: 49%;<br />
height: 100%;<br />
zoom: 1;<br />
display: block;<br />
}<br />
#lightbox-nav-btnPrev { <br />
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float: left;<br />
}<br />
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right: 0; <br />
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font: 10px Verdana, Helvetica, sans-serif;<br />
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margin: 0 auto;<br />
line-height: 1.4em;<br />
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}<br />
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}<br />
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span.reference{<br />
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bottom:10px;<br />
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<span class="sdt_link">Related Links</span><br />
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</span><br />
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<a href="https://2011.igem.org/Team:Missouri_Miners/Papers"> Research Papers</a><br />
<a href="https://2011.igem.org/Team:Missouri_Miners/DiabetesLinks">Diabetes</a><br />
<a href="https://2011.igem.org/Team:Missouri_Miners/GlucoseSensorLinks">Glucose Sensors</a><br />
<a href="https://2011.igem.org/Team:Missouri_Miners/InsulinPumpLinks">Insulin Pumps</a><br />
<a href="https://2011.igem.org/Team:Missouri_Miners/EthicsAndSafetyLinks">Ethics and Safety</a><br />
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</div></div>Eksch9http://2011.igem.org/Team:Missouri_Miners/EthicsAndSafetyLinksTeam:Missouri Miners/EthicsAndSafetyLinks2011-09-27T06:23:58Z<p>Eksch9: </p>
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<h1>Ethics and Safety </h1><br /></div>Eksch9http://2011.igem.org/Team:Missouri_Miners/InsulinPumpLinksTeam:Missouri Miners/InsulinPumpLinks2011-09-27T06:23:37Z<p>Eksch9: </p>
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<h1>Insulin Pumps </h1><br /></div>Eksch9http://2011.igem.org/Team:Missouri_Miners/GlucoseSensorLinksTeam:Missouri Miners/GlucoseSensorLinks2011-09-27T06:23:17Z<p>Eksch9: </p>
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<h1>Glucose Sensors</h1><br /></div>Eksch9http://2011.igem.org/Team:Missouri_Miners/DiabetesLinksTeam:Missouri Miners/DiabetesLinks2011-09-27T06:22:27Z<p>Eksch9: </p>
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<h1> Diabetes </h1><br /></div>Eksch9