IGEM:IMPERIAL/2009/M3/Modelling/Analysis/1: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
 
Line 47: Line 47:


We model the behaviour of our system with 6 differential equations. The first two are concerned with the production of protein cI, which represses killing. The next two tells us the production of restriction enzymes. Equation (5) describes the cell population, while equation (6) tells us the total number of dead cells. This will allow us to relate our model to the [[IGEM:IMPERIAL/2009/M3/Assays/Membrane_staining |live and dead cells assay]] performed.
We model the behaviour of our system with 6 differential equations. The first two are concerned with the production of protein cI, which represses killing. The next two tells us the production of restriction enzymes. Equation (5) describes the cell population, while equation (6) tells us the total number of dead cells. This will allow us to relate our model to the [[IGEM:IMPERIAL/2009/M3/Assays/Membrane_staining |live and dead cells assay]] performed.
In this model, the effects of Dam methylase are not included.


Please click [[IGEM:IMPERIAL/2009/M3/Modelling/Model1 | HERE]] to further explore the models.
Please click [[IGEM:IMPERIAL/2009/M3/Modelling/Model1 | HERE]] to further explore the models.
{{Imperial/09/TemplateBottom}}
{{Imperial/09/TemplateBottom}}

Latest revision as of 03:08, 4 September 2009

<html> <style type="text/css"> /*----------------------------------------------------------------------------*/ /* Structure */ /*----------------------------------------------------------------------------*/

  1. II09_mainWrapper{
   width: 800px;
   align: center;
   border: none;
   margin: 0px auto;
   padding: 0px;

}

/*----------------------------------------------------------------------------*/ /* Content */ /*----------------------------------------------------------------------------*/

  1. II09_content_wrapper {
   width: 550px;
   float: left;
   margin-top: 5px;
   margin-bottom: 5px;
   position:relative;

}

  1. II09_content_boxtop {
   width: 550px;
   height: 10px;
   background:url(http://2009.igem.org/wiki/images/c/c8/II09_content_top.png) no-repeat bottom left;
   position: relative;
   float: left;

}

  1. II09_content_boxbottom {
   width: 550px;
   height: 10px;
   background:url(http://2009.igem.org/wiki/images/b/b4/II09_content_bottom.png) no-repeat top left;
   position: relative;
   float: left;

}

  1. II09_content {
   width: 550px;
   padding: 10px;
   background-color:#ffffff;
   border-left: 1px solid #ccc;
   border-right: 1px solid #ccc;
   position: relative;
   float: left;
   font-size: 9pt;
   text-align: justify;

}

/*----------------------------------------------------------------------------*/ /* Tabs */ /*----------------------------------------------------------------------------*/

.II09_Tabs_Blue {

 float:left;
 width:100%;
 background:#CCCCFF url("http://openwetware.org/images/b/bb/SDbg.gif") repeat-x bottom;
 font-size:93%;
 line-height:normal;
 margin: 0;
 padding: 0;

}

.II09_Tabs_Blue ul {

 margin:0;
 padding:10px 10px 0;
 list-style:none;

} .II09_Tabs_Blue li {

 float:left;
 background:url("http://openwetware.org/images/b/b7/SDleft.gif") no-repeat left top;
 margin:0;
 padding:0 0 0 9px;

}

.II09_Tabs_Blue b {

 float:left;
 display:block;
 background:url("http://openwetware.org/images/f/f8/SDright.gif") no-repeat right top;
 padding:5px 15px 4px 6px;
 text-decoration:none;
 font-weight:bold;
 color:#765;

} </style> </html>

<html> <body> <!--<div id="II09_mainWrapper">--> </html> <html>

<head> <style type="text/css">

  1. II09_Header{
 border-top: 1px solid #7b7b7b;
 border-left: 1px solid #7b7b7b;
 border-right: 1px solid #7b7b7b;
 border-bottom: none;
 background: url(http://2009.igem.org/wiki/images/3/39/II09_Banner.jpg);
 background-position: right -75px;
 width: 783px;
 height: 150px;
 
 margin-left:15px;
 align: right;

} </style> </head>

<body>

<div id="II09_Header"> <center>The E.ncapsulator: the disease killer</center> </div>

</body> </html> <html> <head> <style type="text/css">

  1. II09_Breadcrumb{
  backgrounewd: #7b7b7b;
  background: url(http://2008.igem.org/wiki/images/c/c8/Tudelft_menu_header_bg_1.gif);
  border-top: none;
  border-left: 1px #7b7b7b solid;
  border-right: 1px #7b7b7b solid;
  border-bottom: 1px #7b7b7b solid;
  
  filter:alpha(opacity=100);
  -moz-opacity:1.00;
  opacity:1.0;
  text-color: #000000;
  padding-left: 10px;
  margin-left: 15px;
  width: 773px;

} </style> </head>

<body> <div id="II09_Breadcrumb">

<script type="text/javascript">

var address = document.location.href; var addressSplit = address.split("/");

document.write('<a href="http://2009.igem.org/' +addressSplit[3]+ '">Home (' +addressSplit[3]+ ')</a>');

for (i=4;i<8;i++){

  if (addressSplit[i]!=undefined){
     document.write('<span align="left"><font color=#000000> | </font>');
     document.write('<a href="' +addressSplit[i]+ '">' +addressSplit[i]+ '</a>');
  }

}

  document.write('</span>');

</script> <!--<span align="right">iuvbivb</span>--> </div>

</body> </html> <html> <style type="text/css"> /*----------------------------------------------------------------------------*/ /* Menu */ /*----------------------------------------------------------------------------*/

  1. menu {
   width: 200px;
   margin: 5px 15px 0 15px;
   position: relative;
   float: left;

}

  1. menu ul {
   padding: 0;
   margin: 0 0 10px 0;
   list-style: none;

}

  1. menu li {
   margin:0;

}

  1. menu a {
   display: block;
   width: 185px;
   height: 19px;
   margin-top: 1px;
   margin-left:15px;
   background: url(http://openwetware.org/images/4/46/II09_Menu_ItemHover.png) no-repeat right top;
   padding: 2px 4px;
   text-decoration: none;
   color: #fff;

}

  1. menu a:hover {
   display: block;
   width: 185px;
   height: 19px;
   margin-top: 1px;
   margin-left:15px;
   background: url(http://openwetware.org/images/b/bf/II09_Menu_Item.png) no-repeat right top;
   padding: 2px 4px;
   color: #000;

}

  1. menu .selflink {
   /*font-weight:bold;
   width: 200px;
   border: 1px solid #bbb;
   background: url(http://2008.igem.org/wiki/images/c/c8/Tudelft_menu_header_bg_1.gif) repeat-x bottom;
   color: #111;
   padding: 2px 4px;*/
   display: block;
   width: 200px;
   height: 30px;
   margin-top: 1px;
   border: 1px solid #bbb;
   background: url(http://2008.igem.org/wiki/images/c/c8/Tudelft_menu_header_bg_1.gif) repeat-x left bottom;
   padding: 2px 4px;
   color: #000;

}

  1. menu a.external {
   margin: 0;
   padding: 0;
   width: 156px;

}

.menu_header {

   font-weight:bold;
   width: 200px;
   border: 1px solid #bbb;
   background: url(http://2008.igem.org/wiki/images/c/c8/Tudelft_menu_header_bg_1.gif) repeat-x bottom;
   color: #111;
   padding: 2px 4px;
   text-align:left;

} </style>

<div id="menu"> <ul>

<li><strong class="selflink"><img src="http://2009.igem.org/wiki/images/8/83/II09_Thumb_home.png" width="15%" align="left">Home</strong></li> <li><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2009/Project"> Project Description</a></li> <li><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2009/Major_results"> Major results</a></li> <li><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2009/Biobricks"> Submitted parts</a></li> <li><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2009/Achievements"> Achievements</a></li> </ul>

<ul> <li class="menu_header"><img src="http://2009.igem.org/wiki/images/c/c1/II09_Thumb_m1.png" width="15%" align="left"> Module 1: Protein production</li> <li><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2009/M1"> Overview</a></li> <li><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2009/M1/Genetic"> Genetic circuit</a></li> <li><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2009/M1/Results"> Results</a></li> <li><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2009/M1/Modelling"> Modelling</a></li> </ul>

<ul> <li class="menu_header"><img src="http://2009.igem.org/wiki/images/d/df/II09_Thumb_m2.png" width="15%" align="left">Module 2: Encapsulation &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</li> <li><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2009/M2"> Overview</a></li> <li><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2009/M2/Genetic"> Genetic circuit</a></li> <li><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2009/M2/Results"> Results</a></li> <li><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2009/M2/Modelling"> Modelling</a></li> <li><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2009/M2/Pictures"> Picture gallery</a></li> </ul>

<ul> <li class="menu_header"><img src="http://2009.igem.org/wiki/images/2/25/II09_Thumb_m3.png" width="15%" align="left">Module 3: Genomic neutralisation</li> <li><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2009/M3"> Overview</a></li> <li><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2009/M3/Genetic"> Genetic circuit</a></li> <li><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2009/M3/Results"> Results</a></li> <li><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2009/M3/Modelling"> Modelling</a></li> </ul>

<ul> <li class="menu_header"> Resources</li> <li><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2009/Wetlab"> Wetlab</a></li> <li><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2009/Drylab"> Drylab</a></li> </ul>

<ul> <li class="menu_header"> Ethics project</li> <li><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2009/Ethics"> Overview</a></li> <li><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2009/Ethics/Videos"> Video gallery</a></li> <li><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2009/Ethics/Contribute"> Contribute!</a></li> </ul>

<ul> <li class="menu_header"> Team</li> <li><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2009/Contact"> Contact</a></li> <li><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2009/Team"> Who are we?</a></li> <li><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2009/Friends"> Our friends</a></li> </ul>

<ul> <li class="menu_header"><a href="http://openwetware.org/wiki/Template:Imperial/09/Menu/OLDLinks">More links (from old menu)</a></li> </ul>

</div> </html> <html>


<div id="II09_content_wrapper"> <img src="http://2008.igem.org/wiki/images/a/a1/Tudelft_content_top.png" width="572"> <div id="II09_content">


<script type="text/javascript"> var gaJsHost = (("https:" == document.location.protocol) ? "https://ssl." : "http://www."); document.write(unescape("%3Cscript src='" + gaJsHost + "google-analytics.com/ga.js' type='text/javascript'%3E%3C/script%3E")); </script> <script type="text/javascript"> try { var pageTracker = _gat._getTracker("UA-11206997-1"); pageTracker._trackPageview(); } catch(err) {}</script>

</html> <html> <style type="text/css"> /*----------------------------------------------------------------------------*/ /* Structure */ /*----------------------------------------------------------------------------*/

  1. II09_mainWrapper{
   width: 800px;
   align: center;
   border: none;
   margin: 0px auto;
   padding: 0px;

}

/*----------------------------------------------------------------------------*/ /* Content */ /*----------------------------------------------------------------------------*/

  1. II09_content_wrapper {
   width: 550px;
   float: left;
   margin-top: 5px;
   margin-bottom: 5px;
   position:relative;

}

  1. II09_content_boxtop {
   width: 550px;
   height: 10px;
   background:url(http://2009.igem.org/wiki/images/c/c8/II09_content_top.png) no-repeat bottom left;
   position: relative;
   float: left;

}

  1. II09_content_boxbottom {
   width: 550px;
   height: 10px;
   background:url(http://2009.igem.org/wiki/images/b/b4/II09_content_bottom.png) no-repeat top left;
   position: relative;
   float: left;

}

  1. II09_content {
   width: 550px;
   padding: 10px;
   background-color:#ffffff;
   border-left: 1px solid #ccc;
   border-right: 1px solid #ccc;
   position: relative;
   float: left;
   font-size: 9pt;
   text-align: justify;

}

/*----------------------------------------------------------------------------*/ /* Tabs */ /*----------------------------------------------------------------------------*/

.II09_Tabs_Blue {

 float:left;
 width:100%;
 background:#CCCCFF url("http://openwetware.org/images/b/bb/SDbg.gif") repeat-x bottom;
 font-size:93%;
 line-height:normal;
 margin: 0;
 padding: 0;

}

.II09_Tabs_Blue ul {

 margin:0;
 padding:10px 10px 0;
 list-style:none;

} .II09_Tabs_Blue li {

 float:left;
 background:url("http://openwetware.org/images/b/b7/SDleft.gif") no-repeat left top;
 margin:0;
 padding:0 0 0 9px;

}

.II09_Tabs_Blue b {

 float:left;
 display:block;
 background:url("http://openwetware.org/images/f/f8/SDright.gif") no-repeat right top;
 padding:5px 15px 4px 6px;
 text-decoration:none;
 font-weight:bold;
 color:#765;

} </style> </html>



Assumptions

  • The protein cI is constitutively expressed by the strong promoter J23114.


  • Restriction enzymes DpnII and TaqI
    • At higher temperatures, DpnII and TaqI will not undergo conformation changes or undergo denaturation. Therefore, they will work at optimal levels from 28°C to 42°C.
    • No saturation of enzyme restriction sites will occur. Restriction enzyme concentration will always be in access of the substrate.


  • Cell growth
    • Assume that cell growth prior killing by thermoinduction is not constrained by availability of resources. (This correlates to the classic assumptions for exponential phase)


  • Cell death
    • We assume that the rate of cell death is a function of restriction enzyme concentration at the time. Ie, the system has no memory.
    • More precisely, we assume that each cell has a constant probability of dying due to the action of the restriction enzymes in any unit of time.
    • It is this probability of dying that is modelled as is a function of the internal concentration of restriction enzymes only.
    • A hill function is often chosen because initially, there is very little killing with the increase of temperatures. Then, when threshold temperature is reached, killing is expected to be the dominating feature.



Predictions

  • If the temperatures are below threshold, little restriction enzymes are being produced; hence population of live cells will increase exponentially. (Cell growth is not constrained by resources such as food and space.)


  • If the temperatures are above threshold, the population growth of live cells will be constrained by the concentration of restriction enzyme.


  • Initially, cells will grow exponentially as the restriction enzymes take time to be produced. When enough restriction enzymes are being produced, cell death will begin, and an exponential decrease in cell population will be observed.


  • When we increase the temperature, we produce more restriction enzymes at a faster rate. Hence, at higher temperatures, the exponential decrease in live cell numbers will occur earlier. Furthermore, the maximum live cell population at a given temperature will also be lower with increasing temperatures, because a greater concentration of restriction enzyme means more cell death in the same amount of time.


  • If the lambda cI promoter is strong, there will be a higher rate of cell death than cell growth; we will see a decrease in live cell population. However, if lambda cI promoter is weak, there might not be enough killing, so we will only see a reduction in growth of cell population, ie: a slower increase in population.




The Model

We model the behaviour of our system with 6 differential equations. The first two are concerned with the production of protein cI, which represses killing. The next two tells us the production of restriction enzymes. Equation (5) describes the cell population, while equation (6) tells us the total number of dead cells. This will allow us to relate our model to the live and dead cells assay performed.

In this model, the effects of Dam methylase are not included.

Please click HERE to further explore the models. <html> </div> <img src="http://2008.igem.org/wiki/images/8/89/Tudelft_content_bottom.png" width="572"> </div> <!--</div>--> </body> </html>







































<html> <style type="text/css">

  1. II09_Footer{
 width:763px;
 margin-left: 15px;
 background: url(http://2008.igem.org/wiki/images/c/c8/Tudelft_menu_header_bg_1.gif) repeat-x;
 background-position: bottom;
 padding: 10px;
 border-top: 1px #7b7b7b solid;
 border-left: 1px #7b7b7b solid;
 border-right: 1px #7b7b7b solid;
 border-bottom: 1px #7b7b7b solid;
 position:relative;
 float: left;

} </style>

<body> <div id="II09_Footer"> <a href="http://www.mrgene.com"><img src="http://mrgene.com/portaldata/1/resources/_system/img/logo.gif" alt="Mr. Gene" width="5%"></a> &nbsp; <a href="http://www.geneart.com"><img src="http://www.geneart.com/fileadmin/templates/img/logo.gif" alt="Geneart" width="10%"></a> </div> </body> </html>