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| - | <html>
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| - | <head>
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| | | | |
| - | <!-- affichage des maths -->
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| - | <script type="text/javascript"
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| - | src="http://cdn.mathjax.org/mathjax/latest/MathJax.js?config=TeX-AMS-MML_HTMLorMML">
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| - | </script>
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| - |
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| - | <script type="text/x-mathjax-config">
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| - | MathJax.Hub.Config({
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| - | tex2jax: {inlineMath: [['$','$'], ['\\(','\\)']]}
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| - | });
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| - | </script>
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| - | <script type="text/javascript" src="path-to-mathjax/MathJax.js?config=TeX-AMS-MML_HTMLorMML"></script>
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| - |
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| - |
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| - | <style>
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| - |
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| - | #menubar .left-menu noprint{
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| - | color: white;
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| - | }
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| - |
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| - | .right-menu{
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| - | background: none;
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| - | }
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| - |
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| - |
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| - | #content {
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| - | background: none;
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| - | padding:0;
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| - | width:100%;
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| - | border:0;
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| - | margin-top:Opx;
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| - | }
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| - |
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| - |
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| - |
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| - | #p-logo {
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| - | display:none;
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| - | }
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| - |
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| - |
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| - | #top-section {
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| - | border:none;
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| - | margin: 0 auto;
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| - | height:0px ! important;
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| - | width:100%;
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| - | }
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| - |
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| - | img.bg_team {
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| - | /* Set rules to fill background */
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| - | min-height: 100%;
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| - | min-width: 1024px;
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| - |
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| - | /* Set up proportionate scaling */
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| - | width: 100%;
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| - | height: auto;
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| - |
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| - | /* Set up positioning */
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| - | position: fixed;
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| - | top: 0;
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| - | left: 0;
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| - | z-index:-1;
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| - | }
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| - |
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| - |
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| - |
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| - | #catlinks{
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| - | width:0px;
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| - | height:0px;
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| - | display:none;
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| - | }
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| - | #footer-box{
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| - | width:0px;
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| - | height:0px;
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| - | display:none;
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| - | }
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| - | #search-controls {
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| - | display:none;
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| - | }
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| - |
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| - | #globalWrapper{
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| - | margin:0;
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| - | padding:0;
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| - | border:0;
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| - | }
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| - | #top-section {
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| - | border:none;
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| - | margin: 0 auto;
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| - | height:0px ! important;
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| - | width:100%;
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| - | }
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| - | .firstHeading {
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| - | display:none;
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| - | }
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| - |
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| - |
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| - |
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| - |
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| - | body
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| - | {
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| - | background-image: url("https://static.igem.org/mediawiki/2011/0/04/Fondt.png");
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| - | background-repeat: repeat-x;
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| - |
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| - | background-color: #b4b4b4;
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| - | }
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| - |
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| - |
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| - | #header
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| - | {
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| - | background-image: url("https://static.igem.org/mediawiki/2011/1/11/Haut.png");
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| - | width:985px;
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| - | height:170px;
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| - | margin: auto;
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| - | padding-top: 10px;
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| - | margin-top: 0px;
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| - | }
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| - | #main
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| - | {
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| - | width:980px;
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| - | height:600px;/* A faire varier pour que la barre rouge soit au bonne endroit */
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| - | margin: auto;
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| - | padding-left: 5px;
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| - | }
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| - |
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| - | #maintext
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| - | {
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| - | float: left;
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| - | width:980px;
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| - | }
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| - |
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| - | #menu a{
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| - | color: red;
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| - | }
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| - |
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| - | #hmaint
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| - | {
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| - | color: white;
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| - | font-weight: bolder;
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| - | float: left;
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| - | background-image: url("https://static.igem.org/mediawiki/2011/e/e1/Ulbmaintext.png");
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| - | background-repeat:no-repeat;
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| - | width:985px;
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| - | height: 16px;
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| - | padding-left: 4px;
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| - | }
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| - |
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| - | #maint
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| - | {
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| - | float: left;
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| - | padding: 4px;
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| - | width:980px;
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| - | /* Valeur du height de maintext -16px */
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| - | border: 2px inset #27303e;
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| - | text-align: justify;
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| - | background-color: white;
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| - | }
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| - |
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| - | #facebook{
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| - | margin-left: 707px;
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| - | height:200px;
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| - | }
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| - | #joinus{
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| - | background-repeat: no-repeat;
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| - | padding-left: 4px;
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| - | color: white;
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| - | font-weight: bolder;
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| - | width:274px;
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| - | height:16px;
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| - | background-image: url("https://static.igem.org/mediawiki/2011/6/6d/Ulbfacebook.png");
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| - | }
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| - |
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| - | #module{
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| - | background-color: white;
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| - | width:270px;
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| - | height:400px;
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| - | border: 2px inset #27303e;
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| - | clear:float;
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| - | }
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| - |
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| - | #footer{
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| - | width: 980px;
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| - | height: 201px;
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| - | margin:auto;
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| - | text-align: left;
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| - | padding-left: 5px;
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| - |
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| - | }
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| - |
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| - | #sponsor{
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| - | width: 440px;
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| - | height: 201px;
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| - | float: left;
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| - | }
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| - |
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| - | #sponsort{
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| - | background-repeat: no-repeat;
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| - | padding-left:4px;
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| - | color: white;
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| - | font-weight: bolder;
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| - | width:440px;
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| - | height:16px;
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| - | background-image: url("https://static.igem.org/mediawiki/2011/1/11/Ulbsponsor.png");
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| - | }
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| - |
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| - | #sponsori{
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| - | font-weight: bolder;
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| - | background-image:url("https://static.igem.org/mediawiki/2011/e/ed/Logosponsors.png");
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| - | padding: 4px;
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| - | color: #690115;
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| - | background-color: white;
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| - | width:428px;
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| - | height:175px;
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| - | border: 2px inset #27303e;
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| - | text-align:center;
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| - | }
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| - |
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| - | #photo{
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| - | padding-top: 8px;
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| - | background-color: black;
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| - | background-repeat: no-repeat;
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| - | float: left;
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| - | width:245px;
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| - | height: 195px;
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| - | margin-left: 10px;
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| - | }
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| - |
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| - | #photot
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| - | {
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| - | width:247px;
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| - | height: 16px;
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| - | background-image: url("https://static.igem.org/mediawiki/2011/0/0b/Ulbphoto.png");
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| - | }
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| - |
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| - | #photoi
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| - | {
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| - | width:247px;
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| - | height: 187px;
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| - | background-color: black;
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| - | }
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| - |
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| - | #event{
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| - | float: left;
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| - | margin-left: 10px;
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| - | width: 274px;
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| - | height: 201px;
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| - | }
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| - |
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| - | #nextevent{
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| - | background-repeat: no-repeat;
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| - | padding-left: 4px;
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| - | color: white;
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| - | font-weight: bolder;
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| - | width:274px;
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| - | height:16px;
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| - | background-image: url("https://static.igem.org/mediawiki/2011/a/a8/Ulbnextevent.png");
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| - | }
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| - |
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| - | #eventi{
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| - | padding: 4px;
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| - | color: white
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| - | width:266px;
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| - | height: 175px;
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| - | border: 2px inset #27303e;
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| - | background-color: #690115;
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| - | }
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| - |
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| - | #bas{
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| - | margin-top: 4px;
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| - | background-image: url("https://static.igem.org/mediawiki/2011/3/34/Ulbbas.png");
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| - | background-repeat: repeat-x;
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| - | height: 30px;
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| - | margin: auto;
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| - | }
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| - |
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| - | #basi{
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| - | padding-top: 5px;
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| - | color: white;
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| - | text-align: center;
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| - | font-size: 11px;
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| - | }
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| - |
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| - | #basi a{
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| - | color: white;
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| - | }
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| - |
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| - |
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| - | /*code justine */
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| - |
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| - | #menu a /* pour changer les laids liens bleu soulignÈs*/
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| - | {
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| - | position:relative;
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| - | color: white;
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| - | text-decoration: none;
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| - | text-align: center;
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| - | }
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| - |
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| - | #menu
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| - | {
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| - | position:relative;
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| - | top:126px;
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| - | font-size: 14px;
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| - | color:white;
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| - | font-weight: bolder;
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| - | widht: 900px;
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| - |
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| - |
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| - | }
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| - |
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| - | #menu1 /* mise en forme menu1, plein de choses ‡ virer sur la version dÈfinitive*/
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| - | {
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| - | width:800px; /* version dÈfinitive:virer ca*/
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| - | height: 20px; /* version dÈfinitive:virer ca*/
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| - | font-size: 14px;
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| - | text-align:center;
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| - | }
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| - |
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| - | #home
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| - | {
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| - | float:left;
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| - | position: absolute;
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| - | width: 50px;
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| - | height: 20px;
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| - | text-align:center;
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| - |
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| - | }
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| - |
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| - | #project
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| - | {
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| - | float:left;
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| - | position: relative;
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| - | left: 70px;
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| - | width: 70px;
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| - | height: 15px;
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| - | text-align:center;
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| - |
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| - | }
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| - |
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| - | #modeling
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| - | {
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| - | float:left;
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| - | text-align:center;
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| - | position: relative;
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| - | width: 90px;
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| - | height: 20px;
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| - | left:80px;
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| - | }
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| - |
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| - | #results
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| - | {
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| - | float:left;
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| - | position: relative;
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| - | width: 70px;
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| - | height: 20px;
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| - | left:90px;
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| - | text-align:center;
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| - | }
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| - |
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| - | #parts
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| - | {
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| - | float:left;
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| - | position: relative;
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| - | width: 55px;
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| - | height: 20px;
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| - | left:100px;
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| - | text-align:center;
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| - | }
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| - |
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| - | #human
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| - | {
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| - | float:left;
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| - | position: relative;
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| - | width: 130px;
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| - | height: 20px;
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| - | left:110px;
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| - | text-align:center;
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| - | }
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| - |
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| - | #team
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| - | {
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| - | float:left;
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| - | position: relative;
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| - | width: 85px;
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| - | height: 20px;
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| - | left:110px;
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| - | text-align:center;
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| - | margin : auto;
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| - | }
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| - |
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| - | #team td
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| - | {
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| - | text-align:center;
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| - | }
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| - |
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| - | #sponsors
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| - | {
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| - | float:left;
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| - | position: relative;
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| - | width: 90px;
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| - | height: 20px;
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| - | left:110px;
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| - | text-align:center;
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| - | }
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| - |
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| - | .select
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| - | {
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| - | height:21px; /* si tu modifies hauteur du menu1, il faut modifier ca aussi. Il ne s'adapte pas automatiquement, parce qu'il fait en fonctiond ela taille du texte */
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| - | background-color:rgb(105,1,21);
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| - | }
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| - |
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| - | #menu2
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| - | {
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| - | width:985px;
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| - | height:18px;
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| - | font-size: 12px;
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| - | padding-top: 1px;
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| - | }
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| - |
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| - | #menu2project /* juste pour dÈcaler l'Ècriture des sous-menus. plus facile que de repositionner chaque sous-menu, surtout quand c'ets les mÍmes pour modeling et results*/
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| - | {
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| - | position:relative;
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| - | left: 61px;
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| - | width:807px; /* version dÈfinitive:virer ca*/
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| - | height:18px; /* version dÈfinitive:virer ca*/
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| - | }
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| - |
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| - |
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| - | #menu2results /* juste pour dÈcaler l'Ècriture des sous-menus. plus facile que de repositionner chaque sous-menu, surtout quand c'ets les mÍmes pour modeling et results*/
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| - | {
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| - | position:relative;
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| - | left: 220px;
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| - | width:807px; /* version dÈfinitive:virer ca*/
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| - | height:18px; /* version dÈfinitive:virer ca*/
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| - | }
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| - |
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| - | #menu2modeling
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| - | {
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| - | position:relative;
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| - | left:132px;
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| - | width:888px; /* version dÈfinitive:virer ca*/
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| - | height:18px; /* version dÈfinitive:virer ca*/
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| - | }
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| - |
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| - | #souspindel /* mise en forme du sous-menu "pindel", aussi bien pour modeling que result*/
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| - | {
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| - | position:absolute;
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| - | left:50px;
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| - | height:18px;
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| - | text-align:center;
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| - | width:50px;
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| - | }
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| - |
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| - | #soushelper /* mise en forme du sous-menu "plasmide helper", aussi bien pour modeling que result*/
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| - | {
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| - | position:absolute;
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| - | left:111px;
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| - | height:18px;
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| - | text-align:center;
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| - | width:80px;
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| - | top:0px;
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| - | }
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| - |
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| - | #sousproof /* mise en forme du sous-menu "proof of concept", aussi bien pour modeling que result*/
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| - | {
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| - | position:absolute;
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| - | left:193px;
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| - | height:18px;
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| - | text-align:center;
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| - | width:120px;
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| - | top:0px;
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| - | }
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| - |
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| - |
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| - | .left-menu, .left-menu a {
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| - | color: white;
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| - | left: 0px;
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| - | text-align: left;
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| - | color:transparent;
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| - | text-transform: lowercase;
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| - |
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| - | }
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| - | .left-menu a:hover {
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| - | }
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| - | .left-menu:hover {
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| - | color: white;
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| - | background:none;
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| - | }
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| - | .right-menu, .right-menu a {
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| - | right: 0px;
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| - | text-align: right;
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| - | color: white;
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| - |
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| - | }
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| - | #menubar ul {
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| - | color: white;
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| - | list-style: none;
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| - | }
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| - | #menubar li {
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| - | display: inline;
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| - | position: relative;
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| - | cursor: pointer;
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| - | padding-left: 0px;
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| - | padding-right: 0px;
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| - | }
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| - | .left-menu li a {
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| - | padding: 0px 10px 0px 0px;
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| - | }
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| - | .left-menu .selected {
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| - | # color:white;
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| - | }
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| - | #.left-menu .selected:hover {
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| - | # color:white;
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| - | #}
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| - |
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| - | .left-menu:hover a {
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| - | color: white;
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| - | }
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| - | .right-menu li {
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| - | # background: none;
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| - | }
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| - | .right-menu li a {
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| - | padding: 0px 15px 0px 0px;
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| - | color: white;
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| - | background: none;
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| - | }
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| - | .right-menu li a:hover {
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| - | color: white;
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| - | }
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| - |
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| - | #menu a
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| - | {
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| - | margin-left:20px; /*espace entre les lien*/
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| - | display:block; /*Pour que le lien soit un vrai bloc*/
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| - | float:left; /*Pour que les blocs soient alignés horizontalement (essayez de l'enlever pour voir)*/
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| - | height:30px; /*La même hauteur que notre menu*/
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| - | border:none; /*On enlève les bordures (enfin, on ne les met pas, même s'il n'y en avait pas)*/
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| - | }
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| - |
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| - | #couleur
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| - | {
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| - | padding-left: 5px;
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| - | padding-right:5px;
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| - | background-color: #690115;
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| - | }
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| - |
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| - | #sousm
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| - | {
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| - | clear:left;
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| - | position:relative;
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| - | top:112px;
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| - | margin-left: 135px; /*tu change ici pour le bouger horizontalement*/
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| - | color:white;
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| - | widht: 900px;
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| - | }
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| - | #sousm a
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| - | {
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| - | position:relative;
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| - | color: white;
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| - | text-decoration: none;
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| - | text-align: center;
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| - | margin-left:20px; /*espace entre les lien*/
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| - | display:block; /*Pour que le lien soit un vrai bloc*/
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| - | float:left; /*Pour que les blocs soient alignés horizontalement (essayez de l'enlever pour voir)*/
| |
| - | height:30px; /*La même hauteur que notre menu*/
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| - | border:none; /*On enlève les bordures (enfin, on ne les met pas, même s'il n'y en avait pas)*/
| |
| - | }
| |
| - |
| |
| - | </style>
| |
| - |
| |
| - | </head>
| |
| - |
| |
| - | <div id="header"> <!-- Images de logo -->
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| - | <div id="menu">
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| - | <a href="https://2011.igem.org/Team:ULB-Brussels/modeling/introduction">Introduction</a>
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| - | <a href="https://2011.igem.org/Team:ULB-Brussels/modeling/30">Phase at 30°C</a>
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| - | <a href="https://2011.igem.org/Team:ULB-Brussels/modeling/42">Phase at 42°C</a>
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| - | <a href="https://2011.igem.org/Team:ULB-Brussels/modeling/comparison">Comparison with the Wet Lab work </a>
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| - | <a href="https://2011.igem.org/Team:ULB-Brussels/modeling/conclusion">Conclusion</a>
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| - | Modeling : introduction </div>
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| - | <div id="maint">
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| - | We will consider a population of <em>Escherichia Coli</em>, that possesses the Pindel plasmid, inserted beforehand by electroporation. The replication origin (ORI) of this plasmid is activated by RepA101, produced by the plasmid itself; at $42^\circ$C, this protein (RepA101) becomes rapidly inactive. Also, the plasmid includes a three-genes sequence coding proteins (Red recombinase) inducing the insertion, in the DNA, of a gene "X" (at which we added, as a control, a resistance gene to chloramphenicol). The promotor of this three-genes sequence, Pbad, is activated by arabinose. Pindel includes in addition a coding sequence for flippase, whose promotor is blocked by a thermo-sensible repressor (at $42^\circ$C, the promotor isn't repressed at all, whereas at $30^\circ$C it is almost completely repressed, to $\ldots\%$). Those two sequences follow each other, but are encrypted in opposite directions, which eventually leads to an interference phenomenal. Furthermore, flippase, originally coming from yeast, is entirely active at $30^\circ$C, but is disabled at $42^\circ$C. Red recombinase recognizes the gene sequence to insert (the "X" gene followed by the resistance gene) and insert them in the bacteria's genome, while flippase excise the resistance to chloramphenicol package.
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| - | </p>
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| - | <p>
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| - | At start, we place bacteria possessing Pindel at $30^\circ$C on arabinose. The Red recombinase genes are then expressed because of the presence of arabinose. In parallel, a small amount of flippase is produced: in section (2.1), we will study, by a computer simulation, the interference that occurs during the transcription of those two opposite-oriented genes, which will give us an idea of the probability that the flippase sequence eventually is transcribed. Nevertheless, we do expect that the amount of flippase produced stays low. After a while, those bacteria are electroporated in order to insert the X gene.
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| - | </p>
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| - | <p>
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| - | By testing the resistance to chloramphenicol, we can determine which colony's integrated the X gene, coupled with the resistance gene. We take some of those bacteria and place them at $42^\circ$C on glucose. Flippase is then fully produced, and is able to excise the chloramphenicol-resistance package. At the same time, the RepA101-enzyme, allowing the Pindel plasmid to reproduce is strongly suppressed, and becomes fast completely inactive. In consequence, the total number of plasmids remains constant; and the increase in population will dilute that amount of plasmids among the bacteria, leaving most of them without any plasmid. In the same way, flippase stops being produced, and therefore, by natural degradation, disappears. We thus obtain a population possessing the X gene, but devoid of the chloramphenicol resistance nor the Pindel plasmid.
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| - | </p>
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| - | <p>
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| - | It is this whole process that we will try to modelise here. In our model, we considered a few different parameters, of which we will estimate the values, based on biological considerations. Afterwards, we will analyze the coherence of our predictions together with the results of the experiments, or if necessary, adapt the model itself.
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| - | </p>
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| - | <p>
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| - | Let us begin with a proper definition of the different biological functions we are to study:
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| - | <ul>
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| - | <li>$N$ : total amount of bacteria in the considered population;</li>
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| - | <li>$P$ : average amount of Pindel plasmids per bacterium;</li>
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| - | <li>$E_{tot}$ : average amount of RepA101 enzymes per bacterium;</li>
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| - | <li>$E$ : average amount of RepA101 enzymes active per bacterium;</li>
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| - | <li>$F$ : average amount of flippase per bacterium;</li>
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| - | <li>$G_i (i=1,2,3)$ : average amount of the protein $i$ for the Red recombinase, per bacterium.</li>
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| - | </ul>
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| - | </p>
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| - | <div id="basi">
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| - | iGEM ULB Brussels Team - <a href="mailto:sgerard@ulb.ac.be">Contact us</a>
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