Difference between revisions of "Team:Pasteur Paris/Description"

 
(16 intermediate revisions by 3 users not shown)
Line 1: Line 1:
−
{{Pasteur_Paris}}
+
{{Pasteur_Paris_Parts}}
 +
 
 
<html>
 
<html>
  
−
   <div>
+
<link href='https://fonts.googleapis.com/css?family=Open+Sans:400,300,700' rel='stylesheet' type='text/css'>
−
      <p> [in building]
+
<style type="text/css">
−
      </p>
+
 
−
  </div>
+
 
 +
body{
 +
position: relative;
 +
 +
}
 +
 
 +
 
 +
 
 +
h1 {
 +
        display:block;
 +
font-size: 42px;
 +
        font-family: 'Oswald', Arial, sans-serif;
 +
color:#333;
 +
padding-top:-10px;
 +
        padding-bottom:5%;
 +
        margin-left:0%;
 +
}
 +
 
 +
 
 +
 
 +
h2 {
 +
        display:block;
 +
font-size: 30px;
 +
        padding-bottom:2%;
 +
        color:#17A3B5;
 +
        font-family: 'Oswald', Arial, sans-serif;
 +
padding-top:1%;
 +
        margin-left:0%;
 +
}
 +
 
 +
h3 {
 +
        display:block;
 +
font-size: 25px;
 +
        margin-bottom:3%;
 +
        color:#CD7F32;
 +
        font-family: 'Oswald', Arial, sans-serif;
 +
margin-top:0%;
 +
        margin-left:0%;
 +
}
 +
 
 +
 
 +
h4 {
 +
        display:block;
 +
font-size: 25px;
 +
        margin-bottom:3%;
 +
        color:#CECECE;
 +
        font-family: 'Oswald', Arial, sans-serif;
 +
margin-top:10%;
 +
        margin-left:0%;
 +
}
 +
 
 +
h5 {
 +
        display:block;
 +
font-size: 25px;
 +
        margin-bottom:3%;
 +
        color: #d4ac0d;
 +
        font-family: 'Oswald', Arial, sans-serif;
 +
margin-top:10%;
 +
        margin-left:0%;
 +
}
 +
 
 +
.text1 {
 +
width : 60%;
 +
margin-top:2%;
 +
margin-left:0%;
 +
display : inline-block;
 +
line-height : normal;
 +
text-align: justify;
 +
color:#333;
 +
 
 +
}
 +
 
 +
.text3{
 +
width : 70%;
 +
font-size: 14px;
 +
margin-top:5%;
 +
margin-left:0%;
 +
display : inline-block;
 +
line-height : normal;
 +
text-align: justify;
 +
color:#333;
 +
padding:2% 5% 2% 5%;
 +
border-width:0.5px;
 +
border-style:solid;
 +
border-color:#F0D423 ;
 +
 
 +
}
 +
 
 +
#toparrow{
 +
   position:relative;
 +
  z-index:2000;
 +
  font-size:0;
 +
  margin-top:0%;
 +
  margin-left:90%;
 +
}
 +
 
 +
</style>
 +
<body>
 +
<h1><B>Parts description</B></h1>
 +
 
 +
<div class="text1">
 +
<p>
 +
 
 +
<h3>BBa_K2053000  (for Bronze medal): <i>Thalassiosira pseudonana</i> silaffin kinase (tpSTK1)</h3>
 +
 
 +
<i>T. pseudonana</i> silaffin kinase (tpSTK1) is a central BioBrick part. It was described as an efficient diatom silaffin-phosphorylating enzyme [1]. Since it was shown that phosphorylation of silaffins increased their biosilification activity [2], we hypothesized that phosphorylation of silica-binding peptide from our fusion protein would increase its biosilification properties.</br></br>
 +
To facilitate protein purification, we added a HisTag. To remove the HisTag, we separated the HisTag from the sequence by the TEV cleavage site sequence. </br>
 +
 
 +
<h4>BBa_K2053002 (for Silver medal) : Fusion Si4-cellulose-binding domain-B of domain protein A</h4>
 +
 
 +
In order to functionalize a cellulose-based patch into a biodetection device we designed and created a fusion protein by assembling three parts: the phage displayed silica-binding peptide (Si4) [3],  the <i>Clostridium cellulovorans</i> cellulose-binding domain of cellulose-binding protein A (CBPa) [4], and the B domain of <i>Staphylococcus aureus</i> protein A (BpA) [5]. This fusion protein is used to bind to cellulose-based patch, increase its rigidity by biosilification, and make it a detection device by fixing the Fc fragment of specific antibodies. We used the preexisting iGEM BioBrick parts that we assembled thanks to commonly used flexible linkers [6]. </br></br>
 +
 
 +
To facilitate protein purification, we added a His-Tag composed  of 6 histidines at the N-terminus, or C-terminus. To remove the His-Tag, we inserted a TEV (Tobacco Etch virus) cleavage site downstream from the His-Tag. </br></br>
 +
 
 +
<h5> Fusion cellulose-binding domain of CBP-B domain of protein A (for Gold medal)</h5>
 +
 
 +
We improved previously existing BBa_K863110 and BBa_K103003 BioBricks parts by combining them, creating one part. It is a fusion protein between the <i>Clostridium cellulovorans</i> cellulose-binding domain of cellulose-binding protein A (CBPa) with the B domain of <i>Staphylococcus aureus</i> protein A (BpA). This fusion protein can be used to bind antibodies to cellulose.</br></br>
 +
 
 +
To facilitate protein purification, we added a His-Tag. To remove the His-Tag, we separated the HisTag from the sequence by the TEV cleavage site sequence.</br>
 +
 
 +
</p>
 +
</div>
  
−
  <div>
+
<div class="text3"><p>
−
      <a href="#Trap device">
+
<h2>References: </h2>
−
        <img src="https://photos-1.dropbox.com/t/2/AADNX8fmdVyVzen3fbmBoYF4FidHX3-VhLDx1N-94Zmvqw/12/540076691/png/32x32/1/_/1/2/Trapping%20device.png/EO2yk6kEGCsgAigC/dKNtsq3-2LVzo8VWsg2rRRECdHW8J1eCNi9hroxJqTI?size=1280x960&size_mode=3"/>
+
−
      </a>
+
−
  </div>
+
  
−
  <div>
+
[1] Characterization of an Endoplasmic Reticulum-associated Silaffin Kinase from the diatom <i>Thalassiosira pseudonana</i>, Sheppard V et al, 2010, The Journal of Biological Chemistry</br>
−
      <a href="#Patch">
+
[2] The Role of Proteins in Biosilification, Otzen D, 2012, Scientifica</br>
−
        <img src="https://photos-6.dropbox.com/t/2/AABAue0Ol2z6XVb2bgRzysoAKBtIExDDFp44Jn-gHD83Xw/12/540076691/png/32x32/1/_/1/2/The%20Patch.png/EO2yk6kEGC4gAigC/bPz-aYBfcsS_lukI96eu03gv6t9KTXf-So-WdA2PkQY?size_mode=5/>
+
[3] Silica-Precipitating Peptides Isolated from a Combinatorial Phage Display Peptide Library Naik et al, 2002, Journal of Nanoscience and Nanotechnology</br>
−
      </a>
+
[4] Characterization of the cellulose-binding domain of the <i>Clostridium cellulovorans</i> cellulose-binding protein A, Goldstein MA, 1993, J Bacteriol</br>
−
  </div>
+
[5] Crystallographic refinement and atomic models of a human Fc fragment and its complex with fragment B of protein A from <i>Staphylococcus aureus</i> at 2.9- and 2.8-A resolution, Deisenhofer J, 1981, Biochemistry</br>
 +
[6] Fusion Protein Linkers: Property, Design and Functionality, Xiaoying C, 2013, Advanced Drug Delivery Reviews</br>
  
−
  <div>
+
</br>
−
      <a href="#Data analysis>
+
</p></div>
−
        <img src="https://photos-2.dropbox.com/t/2/AADWvnYPlRcNf1XDNyFTOmvQEHaXcZYC9efZoUSS9GARrQ/12/540076691/png/32x32/1/_/1/2/Data%20analysis.png/EO2yk6kEGC4gAigC/QbFXV7jInyvDMWBNw-xFY-vroaufGngWuB24KowpT7A?size_mode=5/>
+
−
      </a>
+
−
  </div>
+
  
−
  <div>
+
<a id="toparrow" href="#"><img src = "https://static.igem.org/mediawiki/2016/f/f5/Arrow_Design.png" width= 8% alt=""><a/>  
−
      <a href =#Data mapping>
+
−
        <img src="https://photos-3.dropbox.com/t/2/AAAk0NmZUlbZSySly7kpJz16HoBFDqASTj0r-6X6vtdiJg/12/540076691/png/32x32/1/_/1/2/Data%20mapping.png/EO2yk6kEGC4gAigC/FHJVv7nLipspEXYqH5n5C_xplUXXlYi5brfaPsjADt0?size_mode=5
+
−
      </a>
+
−
  </div>
+
  
−
  <div>
 
−
      <h2 id="Trap device">Trap device</h2>
 
−
  </div>
 
  
−
  <div>
 
−
      <h2 id="Patch">Patch</h2>
 
−
  </div>
 
  
−
  <div>
 
−
      <h2 id="Data analysis">Data analysis</h2>
 
−
  </div>
 
  
−
  <div>
+
</body>
−
      <h2 id="Data mapping">Data mapping</h2>
+
−
  </div>
+
  
 
</html>
 
</html>

Latest revision as of 09:34, 14 October 2016