Team:Oxford/why do we need microcompartments
From 2014.igem.org
(Difference between revisions)
Line 1: | Line 1: | ||
+ | {{Team:Oxford/templates/header}} | ||
+ | |||
<html> | <html> | ||
- | |||
- | |||
- | |||
- | + | <style> | |
- | + | .modelling { | |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | . | + | |
- | background: # | + | background: #F9A7B0; |
- | border-radius: | + | border-radius:30px; margin-top:2%; padding-left:2%;padding-right:2%;padding-top:2%;padding-bottom:2%; |
width:96%; | width:96%; | ||
} | } | ||
- | . | + | .wetlab { |
background: #ADD8E6; | background: #ADD8E6; | ||
Line 178: | Line 19: | ||
} | } | ||
- | . | + | .white_news_block1 { |
- | background: # | + | background: #fff; |
- | border-radius: | + | border-radius:15px; margin-top:2%; margin-left: 13%; padding-left:2%;padding-right:2%;padding-top:2%;padding-bottom:2%; |
- | width: | + | width:70%; |
} | } | ||
- | + | .white_news_block2 { | |
- | . | + | |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
background: #fff; | background: #fff; | ||
- | border-radius:15px; margin-top:2 | + | border-radius:15px; margin-top:2%; padding-left:2%;padding-right:2%;padding-top:2%;padding-bottom:2%; |
- | width:96%; | + | width:96%;margin-left:0%; |
} | } | ||
+ | </style> | ||
- | + | <body> | |
- | + | <div class="outer" style="overflow-y: scroll; overflow-x: hidden;"> | |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | <div id="stuff" style="float:left;position:absolute;margin-left:200px;margin-right:100px; margin-top:50px;min-width:645px;"> | |
- | + | <img src="https://static.igem.org/mediawiki/2014/0/0f/Real_Bioremediation.jpg" style="position:absolute; width:100%;z-index:-1; border-radius:15px;"/> | |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | <div style="background-color:#D9D9D9; opacity:0.7; z-index:5; Height:75px; width:100%;font-size:65px;font-family:Helvetica;padding-top:5px; font-weight: 450;margin-top:10px;"> | |
+ | <div style="background-color:white; opacity:0.9; Height:75px; width:100%;margin-top:5px:margin-bottom:5px;font-size:65px;font-family:Helvetica;padding-top:5px; color:#00000; font-weight: 450;"><br><center><font style="opacity:0.7">Why are we using them?</font></center></div> | ||
+ | </div> | ||
+ | <br> | ||
+ | <div style="border-bottom-left-radius:10px;border-bottom-right-radius:10px; padding-left:10px;padding-right:10px;min-width:300px;"> | ||
+ | <img src="https://static.igem.org/mediawiki/2014/d/d5/Realisation_bioremediation.png" style="width:35%;margin-left:33%;margin-top:-50px;"> | ||
+ | </div> | ||
+ | <div style="background-color:white; border-bottom-left-radius:10px;border-radius:10px; padding-left:10px;padding-right:10px;min-width:300px;margin-top:-50px;"> | ||
+ | <a href="#"><img src="https://static.igem.org/mediawiki/2014/5/50/OxigemLabbook.png" style="position:absolute;width:6%;margin-left:84%;margin-top:-13%;z-index:10;"></a> | ||
+ | <a href="#"><img src="https://static.igem.org/mediawiki/2014/a/a4/OxigemProtocols.png" style="position:absolute;width:6%;margin-left:91%;margin-top:-13%;z-index:10;"></a> | ||
+ | <div style="position:absolute;background-color:rgba(255,255,255,0.6);border-radius:15px; z-index:5;margin-top:-18.2%; Height:70px; width:20%;font-size:65px;font-family:Helvetica; font-weight: 450;padding-left:10px;padding-right:10px;padding-top:3px;min-width:170px;margin-bottom:3px;"> | ||
+ | |||
+ | <div style="width:100%;"><font style="font-size:15px;font-weight:500;">Show all:</font></div> | ||
+ | <a href="#showmodelling"><div class="orange_news_block1 showmodelling" style="background: #F9A7B0;border-radius:15px;color:black;float:left;height:40%;width:40%;margin-left:6%;padding-top:5%;margin-top:3px;"><center> | ||
+ | <h1white><font style="font-size:15px;font-weight:500;">Modelling</font> | ||
+ | </div></a> | ||
- | + | <a href="#showwetlab"><div class="orange_news_block1 showwetlab" style="background: #ADD8E6;border-radius:15px;color:black;float:left;height:40%;width:40%;padding-top:5%;margin-left:3%;margin-top:3px;"><center> | |
- | + | <h1white><font style="font-size:15px;font-weight:500;">Wetlab</font> | |
- | + | </div></a> | |
- | + | <br><br><br><br><br> | |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | < | + | |
</div> | </div> | ||
- | <br | + | <br> |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
<h1>Why do we need microcompartments?</h1> | <h1>Why do we need microcompartments?</h1> | ||
- | Using microcompartments, we can boost the bioremediation capacity of E. coli and P. putida in two key ways: by increasing the degradation rate of DCM, and by preventing toxic metabolic intermediates from damaging the cell. How this happens and to what extent we expect this to improve DCMation is described below. | + | Using microcompartments, we can boost the bioremediation capacity of E. coli and P. putida in two key ways: by increasing the degradation rate of DCM, and by preventing toxic metabolic intermediates from damaging the cell. How this happens and to what extent we expect this to improve DCMation is described below.<br><br> |
</div> | </div> | ||
Line 441: | Line 86: | ||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
<div class="row"> | <div class="row"> | ||
- | <a href="# | + | <a href="#show1" class="show" id="show1"><div class="wetlab"> |
- | < | + | <h1white>Increasing the reaction rate</h1white> |
<img src="https://static.igem.org/mediawiki/2014/4/4d/Oxford_plus-sign-clip-art.png" style="float:right;position:relative; width:2%;" /> | <img src="https://static.igem.org/mediawiki/2014/4/4d/Oxford_plus-sign-clip-art.png" style="float:right;position:relative; width:2%;" /> | ||
</div></a> | </div></a> | ||
- | <a href="# | + | <a href="#hide1" class="hide" id="hide1"><div class="wetlab"> |
- | <h1white>Increasing the reaction rate</h1white> | + | <h1white>Increasing the reaction rate</h1white></div></a> |
- | </div></a> | + | |
<div class="list"> | <div class="list"> | ||
- | + | <div class="white_news_block2"> | |
- | + | ||
- | <div class=" | + | |
<h1>Increased concentration of metabolic enzymes</h1> | <h1>Increased concentration of metabolic enzymes</h1> | ||
Proteins can be targeted into microcompartments via an 18-amino acid leader sequence that forms a well-defined helical structure (Lawrence AD, 2014). We added this leader sequence to dcmA in both E. coli and P. putida. The resulting accumulation of dcmA in the microcompartments leads to an increased local concentration of the metabolic enzyme. This allows more rapid degradation of DCM, which can diffuse freely through the plasma membrane and into microcompartments. | Proteins can be targeted into microcompartments via an 18-amino acid leader sequence that forms a well-defined helical structure (Lawrence AD, 2014). We added this leader sequence to dcmA in both E. coli and P. putida. The resulting accumulation of dcmA in the microcompartments leads to an increased local concentration of the metabolic enzyme. This allows more rapid degradation of DCM, which can diffuse freely through the plasma membrane and into microcompartments. | ||
Line 488: | Line 117: | ||
</div> | </div> | ||
</div> | </div> | ||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
Line 497: | Line 134: | ||
<div class="row"> | <div class="row"> | ||
- | <a href="# | + | <a href="#show2" class="show" id="show2"><div class="modelling"> |
<h1white>Introduction to stochastic diffusion models</h1white> | <h1white>Introduction to stochastic diffusion models</h1white> | ||
<img src="https://static.igem.org/mediawiki/2014/4/4d/Oxford_plus-sign-clip-art.png" style="float:right;position:relative; width:2%;" /> | <img src="https://static.igem.org/mediawiki/2014/4/4d/Oxford_plus-sign-clip-art.png" style="float:right;position:relative; width:2%;" /> | ||
</div></a> | </div></a> | ||
- | <a href="# | + | <a href="#hide2" class="hide" id="hide2"><div class="modelling"> |
- | <h1white>Introduction to stochastic diffusion models</h1white> | + | <h1white>Introduction to stochastic diffusion models</h1white></div></a> |
- | </div></a> | + | |
<div class="list"> | <div class="list"> | ||
- | <div class=" | + | <div class="white_news_block2"> |
<h1>Stochastic Reaction-Diffusion models</h1> | <h1>Stochastic Reaction-Diffusion models</h1> | ||
Because of the relatively small number of molecules we are expecting to have in our cells (≈10^5 enzymes per cell and 10^3 enzymes per microcompartment), we developed stochastic reaction-diffusion models to predict the distribution of formaldehyde within the system. These stochastic models build in an element of randomness that reflects the nature of diffusion for systems with few elements in a way that deterministic relationships such as Fick’s law do not. | Because of the relatively small number of molecules we are expecting to have in our cells (≈10^5 enzymes per cell and 10^3 enzymes per microcompartment), we developed stochastic reaction-diffusion models to predict the distribution of formaldehyde within the system. These stochastic models build in an element of randomness that reflects the nature of diffusion for systems with few elements in a way that deterministic relationships such as Fick’s law do not. | ||
Line 533: | Line 169: | ||
</div> | </div> | ||
</div> | </div> | ||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
Line 540: | Line 208: | ||
<div class="row"> | <div class="row"> | ||
- | <a href="# | + | <a href="#show3" class="show" id="show3"><div class="wetlab"> |
<h1white>Microcompartment/rate of collision models</h1white> | <h1white>Microcompartment/rate of collision models</h1white> | ||
<img src="https://static.igem.org/mediawiki/2014/4/4d/Oxford_plus-sign-clip-art.png" style="float:right;position:relative; width:2%;" /> | <img src="https://static.igem.org/mediawiki/2014/4/4d/Oxford_plus-sign-clip-art.png" style="float:right;position:relative; width:2%;" /> | ||
</div></a> | </div></a> | ||
- | <a href="# | + | <a href="#hide3" class="hide" id="hide3"><div class="wetlab"> |
- | <h1white>Microcompartment/rate of collision models</h1white> | + | <h1white>Microcompartment/rate of collision models</h1white></div></a> |
- | </div></a> | + | |
<div class="list"> | <div class="list"> | ||
- | + | <div class="white_news_block2"> | |
<h1>Modelling the effect of introducing a microcompartment on reaction rate</h1> | <h1>Modelling the effect of introducing a microcompartment on reaction rate</h1> | ||
Our main approach to reducing the accumulation of toxic intermediates in the DCM degradation pathway is the expression of Pdu microcompartments in our bacteria that will contain high concentrations of both the DcmA enzyme and FdhA enzyme. In doing so, we increase the likelihood of a formaldehyde molecule produced by the action of DcmA encountering an FdhA enzyme before it leaves the microcompartment thus reducing the accumulation of formaldehyde. | Our main approach to reducing the accumulation of toxic intermediates in the DCM degradation pathway is the expression of Pdu microcompartments in our bacteria that will contain high concentrations of both the DcmA enzyme and FdhA enzyme. In doing so, we increase the likelihood of a formaldehyde molecule produced by the action of DcmA encountering an FdhA enzyme before it leaves the microcompartment thus reducing the accumulation of formaldehyde. | ||
Line 588: | Line 255: | ||
</div> | </div> | ||
</div> | </div> | ||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
Line 601: | Line 282: | ||
<div class="row"> | <div class="row"> | ||
- | <a href="# | + | <a href="#show4" class="show" id="show4"><div class="modelling"> |
- | < | + | <h1white>Reducing accumulation of toxic intermediates</h1white> |
<img src="https://static.igem.org/mediawiki/2014/4/4d/Oxford_plus-sign-clip-art.png" style="float:right;position:relative; width:2%;" /> | <img src="https://static.igem.org/mediawiki/2014/4/4d/Oxford_plus-sign-clip-art.png" style="float:right;position:relative; width:2%;" /> | ||
</div></a> | </div></a> | ||
- | <a href="# | + | <a href="#hide4" class="hide" id="hide4"><div class="modelling"> |
- | <h1white>Reducing accumulation of toxic intermediates</h1white> | + | <h1white>Reducing accumulation of toxic intermediates</h1white></div></a> |
- | </div></a> | + | |
<div class="list"> | <div class="list"> | ||
- | + | <div class="white_news_block2"> | |
- | + | ||
- | <div class=" | + | |
Formaldehyde is genotoxic because it forms protein-DNA and DNA-DNA cross-links. This interferes with DNA replication, repair, and transcription, as well as other processes. It has previously been reported that expression of dcmA in E. coli in the presence of DCM leads to cell death. We hypothesise to be caused by the formaldehyde that is released in the dcmA reaction. The localisation of dcmA into microcompartments means that the formaldehyde is also only locally produced within the microcompartment, thus preventing it from causing damage to DNA. | Formaldehyde is genotoxic because it forms protein-DNA and DNA-DNA cross-links. This interferes with DNA replication, repair, and transcription, as well as other processes. It has previously been reported that expression of dcmA in E. coli in the presence of DCM leads to cell death. We hypothesise to be caused by the formaldehyde that is released in the dcmA reaction. The localisation of dcmA into microcompartments means that the formaldehyde is also only locally produced within the microcompartment, thus preventing it from causing damage to DNA. | ||
Line 635: | Line 313: | ||
</div> | </div> | ||
</div> | </div> | ||
- | |||
- | |||
- | |||
- | |||
Line 653: | Line 327: | ||
<div class="row"> | <div class="row"> | ||
- | <a href="# | + | <a href="#show5" class="show" id="show5"><div class="wetlab"> |
<h1white>Modelling formaldehyde concentration against time</h1white> | <h1white>Modelling formaldehyde concentration against time</h1white> | ||
<img src="https://static.igem.org/mediawiki/2014/4/4d/Oxford_plus-sign-clip-art.png" style="float:right;position:relative; width:2%;" /> | <img src="https://static.igem.org/mediawiki/2014/4/4d/Oxford_plus-sign-clip-art.png" style="float:right;position:relative; width:2%;" /> | ||
</div></a> | </div></a> | ||
- | <a href="# | + | <a href="#hide5" class="hide" id="hide5"><div class="wetlab"> |
- | <h1white>Modelling formaldehyde concentration against time</h1white> | + | <h1white>Modelling formaldehyde concentration against time</h1white></div></a> |
- | </div></a> | + | |
<div class="list"> | <div class="list"> | ||
- | + | <div class="white_news_block2"> | |
<h1>Predicted effect of microcompartments on formaldehyde concentration</h1> | <h1>Predicted effect of microcompartments on formaldehyde concentration</h1> | ||
Having deduced that microcompartments will increase the rate at which intermediate compounds are degraded, the next step was to create a simulation that would predict how microcompartments would therefore affect the concentration of formaldehyde molecules in the system. To do so, we created a 1-D simulation in which we started with a fixed number of molecules while constraining degradation and production of a species to within pre-defined spatial limits- representing the fact that both these phenomena can only occur in the microcompartment in our actual system. | Having deduced that microcompartments will increase the rate at which intermediate compounds are degraded, the next step was to create a simulation that would predict how microcompartments would therefore affect the concentration of formaldehyde molecules in the system. To do so, we created a 1-D simulation in which we started with a fixed number of molecules while constraining degradation and production of a species to within pre-defined spatial limits- representing the fact that both these phenomena can only occur in the microcompartment in our actual system. | ||
Line 696: | Line 369: | ||
- | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | <div class="white_news_block2"> | ||
<img src="https://static.igem.org/mediawiki/2014/7/76/Oxford_Micro_title.png" style="float:right;position:relative; width:48%;" /> | <img src="https://static.igem.org/mediawiki/2014/7/76/Oxford_Micro_title.png" style="float:right;position:relative; width:48%;" /> | ||
Line 726: | Line 409: | ||
<br><br><br> | <br><br><br> | ||
Oxford iGEM 2014 | Oxford iGEM 2014 | ||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
</div> | </div> | ||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | < | + | <br> |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
</body> | </body> | ||
</html> | </html> | ||
+ | |||
+ | {{Team:Oxford/templates/footer}} |
Revision as of 22:29, 11 October 2014