Difference between revisions of "Team:Pasteur Paris/TrainingThomas2"

 
Line 4: Line 4:
 
   <style type="text/css">
 
   <style type="text/css">
 
     #runningchip {
 
     #runningchip {
−
       left: 605px;
+
       left: 845px;
 
     }
 
     }
−
     #hp_small {
+
     #labwork_small {
 
       background-color: #292929;
 
       background-color: #292929;
 
     }
 
     }
Line 13: Line 13:
 
           overflow: hidden;
 
           overflow: hidden;
 
           margin: 0 auto;
 
           margin: 0 auto;
 +
          z-index:-1;
 +
            position: relative;
 +
            text-align: center;
 +
            color: white;
 
     }
 
     }
−
     @media screen and (max-width: 760px) {
+
     @media screen and (max-width: 480px) {
 
     /*#wrapper {
 
     /*#wrapper {
 
           width: 100%;
 
           width: 100%;
Line 28: Line 32:
 
     }
 
     }
  
−
    ul li{
 
−
      list-style: disc;
 
−
    }
 
 
   </style>
 
   </style>
  
 
   <div id="bannerchanged">
 
   <div id="bannerchanged">
−
                 <img class="banner-img" src="https://static.igem.org/mediawiki/2018/6/64/T--Pasteur_Paris--Banner_Results.jpg">
+
                 <img class="banner-img" src="https://static.igem.org/mediawiki/2018/1/19/T--Pasteur_Paris--Banner_Team.jpg">
 
         </div>
 
         </div>
−
    <h1></h1>
 
−
  </div>
 
  
 
   <div id="GeneralContent">
 
   <div id="GeneralContent">
−
    <div id="index" class="block">
 
−
      <div id="indexContent">
 
−
        <p><a href="#Reconnect" class="link">Reconnect Nerves</a></p>
 
−
        <p><a href="#Fight" class="link">Fight Infections</a></p>
 
−
        <p><a href="#Kill" class="link">Kill Switch</a></p>
 
−
        <p><a href="#Membrane" class="link">Membrane</a></p>
 
−
      </div>
 
−
      <div id="indexRight">
 
−
        <div id="littleButton"></div>
 
−
      </div>
 
−
    </div>
 
 
     <div id="MainContent">
 
     <div id="MainContent">
  
−
<!-- First Onglet Reconnect nerves-->
 
−
                <div class="block full bothContent">
 
−
                    <div class="block dropDown" id="Reconnect">
 
−
                        <h4>RECONNECT NERVES: <span>Click to see more</span></h4>
 
−
                    </div>
 
−
 
−
                    <div class="block hiddenContent">
 
−
                        <span class="closeCross"><img src="https://static.igem.org/mediawiki/2018/6/67/T--Pasteur_Paris--CloseCross.svg"></span>
 
−
          <div class="block title">
 
−
                <h1>RECONNECT NERVES</h1>
 
−
            </div>
 
  
−
        <div class="block half">
+
<div class="block title">
−
                <h4 style="text-align: left;">DNA assembly</h4><br><br>
+
<h1> Let's meet our team! </h1>
−
                <p>The <b>sequence</b> we designed codes for two different proteins: <b>proNGF (Nerve Growth Factor)</b> and <b>TEV protease</b> (from Tobacco Etch Virus). These two proteins are fused in C-terminal with a signal peptide for <i>E. coli</i> Type I Secretion System which consists in the last 60 amino-acids of HaemolysinA (<b>HlyA</b>). Each coding sequence is separated from the signal peptide by the cleavage sequence for TEV, in order to get the protein without its signal peptide (Figure 3).</p>
+
−
            </div>
+
−
            <div class="block half">
+
−
                <img src="https://static.igem.org/mediawiki/2018/8/80/T--Pasteur_Paris--ProNGFandTEVproduction.png">
+
−
                <div class="legend"><b>Figure 1: </b>proNGF and TEV production cassette </div>
+
−
            </div>
+
−
            <div class="block full">
+
−
                <p>This DNA construct was ordered in two parts, named Seq1 (1096 bp) and Seq2 (1153 bp) in commercial plasmids pEX-A258 from gene synthesis. Seq1 and Seq2 were amplified in competent <i>E. coli</i> DH5-<FONT face="Raleway">α</FONT>.  After bacteria culture and plasmid DNA extraction, we digested commercial vectors with restriction enzymes (<b>NheI</b> and <b>BamHI</b> for Seq1, <b>MscI</b> and <b>HindIII</b> for Seq2). We extracted the inserts from the gel and performed a ligation by using specific overlaps into <b>linearized pET43.1a</b> for proNGF expression and into <b>pSB1C3</b> for iGEM sample submission.<br>We repeated the procedure (transformation in <i>E. coli</i>  Stellar competent cells, bacteria culture, plasmid DNA extraction, digestion) and we proved that our vector pet43.1a contained Seq1 and Seq2 (Figure 2) and that pSB1C3 contained Seq1 and Seq2 (Figure 3) after digestion and DNA electrophoresis. Plasmid DNA of pSB1C3 construction was purified and sent for sequencing (Figure 4).</p>
+
−
            </div>
+
−
            <div class="block half">
+
−
                <img src="https://static.igem.org/mediawiki/2018/7/7c/T--Pasteur_Paris--gelNGFpET.png">
+
−
                <div class="legend"><b>Figure 2: </b> Agarose 1% gel after electrophoresis of digested pET43.1 containing Seq1 and Seq2 (Bba_K2616000) with NdeI. Colonies 6, 9, 10 ,11, 15 have the correct construction.</div>
+
 
</div>
 
</div>
−
            <div class="block half">
 
−
                <img src="https://static.igem.org/mediawiki/2018/e/ef/T--Pasteur_Paris--gelNGFpSB1C3.png">
 
−
                <div class="legend"><b>Figure 3: </b> Agarose 1% gel after electrophoresis of digested pSB1C3 containing Seq1 and Seq2 (Bba_K2616000) with EcoRI/PstI. Colonies 3, 7 and 8 have the correct construction.</div>
 
−
            </div>
 
  
 +
<section class="cf team-container">
 +
  <h1 class="team-h1">Students</h1>
  
−
            <div class="block full">
 
−
                <p>Alignment of <b>Sequencing</b> Results then confirmed that pSB1C3 contained Seq1 and Seq2, <b><a href="">BBa_K2616000 </a></b>. </p>
 
−
            </div>
 
−
            <div class="block two-third center">
 
−
                <img src="https://static.igem.org/mediawiki/2018/e/ee/T--Pasteur_Paris--Sequencing_proNGF.PNG">
 
−
                <div class="legend"><b>Figure 4: </b> Alignment of sequencing results for BBa_K2616000. Sequencing perform in pSB1C3 and three primers were designed (FOR1, FOR2, FOR3) to cover the whole sequence. Image from Geneious. </div>
 
−
            </div>
 
−
            <div class="block full">
 
−
                <p>The construction was successfully assembled. On Figure 4, mismatches are visible which correspond to the reduced precision of sequencing after 600 bp. To avoid this lack of precision, we used three different primers, allowing us to cover the whole sequence without mistakes. As visible, the mismatches are only present at the extremities of each primer sequencing. </p>
 
−
            </div>
 
  
−
        <div class="block full">
+
  <!-- member-->
−
                <h4 style="text-align: left;">proNGF characterization and purification</h4><br><br>
+
<div class="block title"> <h2>Etienne Axelos</h2> </div>
 +
    <div class="team-member">
 +
    <div class="block half">
 +
    <img class="team-photo" src="https://static.igem.org/mediawiki/2018/0/0e/T--Pasteur_Paris--Team_Members_Etienne.jpg">
 +
    </br></br></br><a class="linkedIn" href=""><content >LinkedIn</content></a></div>
 +
      <div class="block half">
 +
      <content>Etienne is one of our designers. He studies at the ENSCI-les Ateliers in Paris. He is always wearing black and fluorescent colors, which is very convenient because you can’t miss him. He is also a big music fan, and has special skills in rock’n’roll dance (very impressive!). As you can imagine, he is one of our funniest team members, you are never bored with him. But he is as funny as he is a hard worker, and we can always count on his help. </br> <i>By Emma</i></content>
 +
</div>
 +
    </div>
 +
<div class="block full">
  
−
    <p> Our chassis is <b><i>Escherichia coli </i>BL21(DE3) pLysS</b>, a specific strain dedicated to producing high amounts of desired proteins under a T7 promoter. Thus, we co-transformed our bacteria with <b><a href="">BBa_K2616000 </a></b> and pVDL 9.3, generously provided by Dr. Victor de Lorenzo, from Centro Nacional de Biotecnologia of Madrid, bearing HlyB and HlyD (Type I secretion system) sequences, in order to get a chance to secrete NGF out of the cell.<br><br>
+
</div>
−
Bacteria were grown at a large scale (800 mL), and proNGF expression was induced with 0.1 mM IPTG for 2 hours at 37°C. <br><br>
+
 
−
We tried to achieve His-tagged proNGF purification using Ni-NTA affinity purification column. We eluted our protein using a gradient of imidazole-containing buffer and one peak was detected.<br><br></p>
+
  
−
            </div>
+
    <!-- member-->
−
            <div class="block two-third center">
+
<div class="block title"> <h2>Alice Dejoux</h2> </div>
−
                <img src="https://static.igem.org/mediawiki/2018/6/69/T--Pasteur_Paris--ResultsFPLC.png">
+
    <div class="team-member">
−
                <div class="legend"><b>Figure 5: </b>FPLC proNGF purification with ÄKTA pure (General Electric) Ni-NTA column was equilibrated with buffer A (50 mM Tris, pH 7.4, 200 mM NaCl). Supernatant of lyzed bacteria was introduced through the column. Washing with 5% of buffer B. Elution by buffer B gradient (buffer A + imidazole 250 mM). UV absorbance at 280nm is shown in blue, conductivity in red, and concentration of buffer B in green. </div>
+
<div class="block half">
−
            </div>
+
      <content> Calm, determined, attentive and engaged: that’s how we see Alice. This young Parisian born in Marseille is currently studying at the prestigious school Chimie Paristech. Talented and always smiling she was one of the leaders of the Parisian meet up organisation and she also attended the Mediterranean meet up in Marseille. Her scientific experience made her show great involvement and perseverance in the lab while doing the modeling of the project. She is part of the 23 strong links of the team iGEM Paris Pasteur 2018. </br><i> By Florian</i></content>
−
            <div class="block full">
+
−
                <p>We analyzed bacterial lysate and purification fractions using SDS-PAGE electrophoresis and Mass spectrometry. </p>
+
−
            </div>
+
−
            <div class="block half">
+
−
                <img src="https://static.igem.org/mediawiki/2018/5/53/T--Pasteur_Paris--SDSPage.png">
+
−
 
+
−
                <div class="legend"><b>Figure 6: </b>SDS-PAGE gel Bis-Tris 4-12% of bacterial lysate and proNGF purification fraction by SDS-PAGE.
+
−
</div>
+
−
            </div>
+
−
            <div class="block half">
+
−
<p> The proNGF purification using NiNTA column is not conclusive. Many proteins are found on elution fractions. His-tagged proNGF fused to HlyA export signal should be found at 33 kDa while the proNGF cleaved by TEV protease should be found at 27 kDa. We finally analyzed five gel bands of the FPLC flow-through (lane 2, Figure 6) by mass spectrometry, by LC/MS/MS, to verify the presence of proNGF.</p>
+
 
</div>
 
</div>
 
<div class="block half">
 
<div class="block half">
−
<p>According to Figure 7, proNGF pattern are found on each lane sent to mass spectrometry. The major amount is found on fraction 5, corresponding to 33 kDa, at this molecular weight, the proNGF is still fused to the signal export. The TEV protease, 34 kDa fused to signal export and 28 kDa cleaved from the signal export are found. </p>
+
      <img class="team-photo" src="https://static.igem.org/mediawiki/2018/8/89/T--Pasteur_Paris--Team_Members_Alice.jpg">
 +
      </br></br></br><a class="linkedIn" href="https://www.linkedin.com/in/alice-dejoux-bb2503150/"><content >LinkedIn</content></a></div>
 +
 
 +
    </div>
 +
<div class="block full">
 +
 
 
</div>
 
</div>
  
−
            <div class="block half">
 
−
                <img src="https://static.igem.org/mediawiki/2018/c/cd/T--Pasteur_Paris--distribution.png">
 
−
                <div class="legend"><b>Figure 7: </b>Distribution of proNGF and TEV protease by gel fractions after mass spectrometry analysis. </div>
 
−
            </div>
 
  
−
            <div class="block full">
+
 
−
<p>Analysis of Fraction 5 of the gel shows our protein proNGF is present but is still bound to its signal peptide HlyA. (Figure 8) Mass spectrometry spectrum of Peptide A, IDTACVCVLSR, from proNGF sequence is shown in Figure 9. Mass spectrometry spectrum of Peptide B, IISAAGSFDVKEER from fused HlyA signal export is shown in Figure 9. The presence of mass spectrometry identified peptides corresponding to the fusion of proNGF and HlyA indicate some proNGF uncleaved from the signal export</p>
+
      <!-- member-->
 +
<div class="block title"> <h2>Claire Dumont</h2> </div> 
 +
    <div class="team-member">
 +
<div class="block half">
 +
      <img class="team-photo" src="https://static.igem.org/mediawiki/2018/3/34/T--Pasteur_Paris--Team_Members_Claire.jpg"> </br></br></br><a class="linkedIn" href="https://www.linkedin.com/in/claire-dumont-55a41910b"><content >LinkedIn</content></a></div>
 +
      <div class="block half">
 +
      <content> Claire is one of our three marvelous designers from ENSCi les Ateliers. But that’s not it! She is also the key to keeping calm and focused the other two designers, who are more extroverted. Yet, she still loves to make movie references and silly jokes. In the team, she works very hard to create amazing graphics and she co-developed our final 3D-printed interface. It wouldn’t be an easy task to count the hours she spent on Rhino software to achieve her great results. </br> <i>By Jonathan</i></content>
 
</div>
 
</div>
 +
    </div>
  
−
            <div class="block two-third center">
+
<div class="block full">
−
                <img src="https://static.igem.org/mediawiki/2018/b/ba/T--Pasteur_Paris--Align_sequence_Mass.png">
+
−
                <div class="legend"><b>Figure 8: </b>Alignment sequences of proNGF fused to HlyA export signal and peptides identified by mass spectrometry. In light blue peptides that match proNGF amino acids sequence. In light yellow, peptides that match HlyA signal export. Sequence has been annotated to match corresponding protein amino acid sequences : In orange His tagged proNGF, in red TEV protease cleaving site, in rose HlyA signal export.</div>
+
−
            </div>
+
  
−
            <div class="block two-third center">
+
</div>
−
                <img src="https://static.igem.org/mediawiki/2018/2/20/T--Pasteur_Paris--massspec.png">
+
−
                <div class="legend"><b>Figure 9: </b>Mass spectrometry spectrum. A) Peptide identified corresponding to proNGF. B) Peptide identified corresponding to the fusion of proNGF and HlyA export signal. </div>
+
−
            </div>
+
  
−
            <div class="block full">
 
−
                <p>The proNGF did not seem to be retained on the affinity column. We performed batch purification using NiNTA beads under native and partial denaturing conditions (Urea 2 M) followed by Western Blot analysis with immunodetection through Anti-His Antibodies Alexa Fluor 647. (Figure 10) Detection of His-tag in the pellet supernatant of induced BL21 with 1 mM IPTG and flow through when partially denatured.</p>
 
−
<p> His-tagged proNGF was not retained on NiNTA beads. N-terminal His tag may be hidden in the protein fold. Consequently, we did not manage to purify the proNGF.
 
−
</p></div>
 
  
−
            <div class="block two-third center">
+
      <!-- member--> 
−
                <img src="https://static.igem.org/mediawiki/2018/5/56/T--Pasteur_Paris--WBproNGF.png">
+
<div class="block title"> <h2>Antoine Ehret</h2> </div>
−
                <div class="legend"><b>Figure 10: </b>Western Blot analysis of batch purification proNGF under native and partial denaturing conditions. </div>
+
    <div class="team-member">
−
            </div>
+
<div class="block half">
 +
      <content> He is one of our dearest physicists studying at ESPCI Paris. He may seem like a shy person, but believe us: he is a man capable of wonders and knows a thing or two about inspirationnal speechs. In fact, you could consider him as our never ending source of encouragement. Each word that comes out of him fills you with determination. " You have to push back your limits each and everyday!" </br> <i>By Aymen</i></content>
 +
    </div>
 +
<div class="block half">
 +
      <img class="team-photo" src="https://static.igem.org/mediawiki/2018/2/20/T--Pasteur_Paris--Team_Members_Antoine.jpg"> </br></br></br><a class="linkedIn" href="https://www.linkedin.com/in/antoine-ehret-303219161
 +
"><content >LinkedIn</content></a></div>
 +
</div>
  
−
            <div class="block separator-mark"></div>
+
<div class="block full">
−
                    </div>
+
  
−
        <div class="block full" style="background-color: transparent;">
+
</div>
−
                <p><i>Achievements: </i><br>
+
−
                    <ul style="text-align: left;">
+
−
                        <li>Successfully cloned a part coding for secretion of NGF in pET43.1a and iGEM plasmid backbone pSB1C3, creating a new composite part <a href=BBa_K2616000 "http://parts.igem.org/Part:BBa_K2616000"> BBa_K2616000</a> </li>
+
−
                        <li>Successfully sequenced <a href=BBa_K2616000 "http://parts.igem.org/Part:BBa_K2616000"> BBa_K2616000</a> in pSB1C3 and sent to iGEM registry </li>
+
−
                        <li>Successfully co-transform E. coli with plasmid secreting NGF and plasmid expressing the secretion system, creating bacteria <b>capable of secreting NGF</b> in the medium</li>
+
−
                        <li>Successfully characterized production of NGF thanks to mass spectrometry</li>
+
−
                        <li>Successfully <b>observe axon growth</b> in microfluidic chip in presence of commercial NGF</li>
+
−
                    </ul><br></p>
+
−
                    <p><i>Next steps:</i><br>
+
−
                    <ul style="text-align: left;">
+
−
                        <li><b>Purify</b> secreted NGF, and characterize its effects on neuron growth thanks to our microfluidic device </li>
+
−
                        <li><b>Global proof of concept</b> in a microfluidic device containing neurons in one of the chamber, and our engineered bacteria in the other</li>
+
−
                    </ul>
+
−
                </p>
+
−
                    </div>
+
−
                    </div>
+
  
−
<!-- Second Onglet Fight infections-->
+
    <!-- member-->
−
                <div class="block full bothContent">
+
<div class="block title"> <h2>Andreas Fyrillas</h2> </div>
−
                    <div class="block dropDown" id="Fight">
+
    <div class="team-member">
−
                                      <h4>FIGHT INFECTIONS : Click to see more</h4>
+
<div class="block half">
 +
      <img class="team-photo" src="https://static.igem.org/mediawiki/2018/7/7c/T--Pasteur_Paris--Team_Members_Andreas.jpg"></br></br></br><a class="linkedIn" href="https://www.linkedin.com/in/andreas-fyrillas-460732163/"><content >LinkedIn</content></a></div>
 +
    <div class="block half">
 +
      <content> Behind his calm and hard-working look, Andreas Fyrillas will not hesitate to throw a punchline at you. His studies at the engineering school ESPCI Paris give him the skills to be an expert on microfluidic chips. Not only does he work the oscilloscope like no other, but he also has a secret talent in the kitchen that he uses to delight us from time to time. Apart from coding the wiki, he enjoys sharing his self-made dubstep music with us. </br><i> By Alice</i></content>
 +
    </div>
  
−
                        </div>
+
<div class="block full">
  
−
                    <div class="block hiddenContent">
+
</div>
−
                        <span class="closeCross"><img src="https://static.igem.org/mediawiki/2018/6/67/T--Pasteur_Paris--CloseCross.svg"></span>
+
  
−
            <div class="block title" id="Fight">
+
  <!-- member--> 
−
                <h1>FIGHT INFECTIONS</h1><br>
+
<div class="block title"> <h2>Aymen Ghaneme</h2> </div>
−
            </div>
+
    <div class="team-member">
−
           
+
<div class="block half">
−
            <div class="block full">
+
      <content> Aymen, currently a second-year student at CentraleSupelec, is a smiling Morrocan who is always in for a break and above all a great fruit sugar-free ice cream (in fact, after a while, we realized the break is only a pretext for the icecream). However, when he gets sufficient sleep, this hyper-active guy can use all his mathematical abilities on making models and will work for hours to solve any software bug. </br> <i>By Thomas</i></content>
−
            <h2>RIP Secretion <a href="http://parts.igem.org/Part:BBa_K2616001"> BBa_K2616001</a></h2><br><br>
+
    </div>
 +
<div class="block half">
 +
      <img class="team-photo" src="https://static.igem.org/mediawiki/2018/f/fa/T--Pasteur_Paris--Team_Members_Aymen.jpg"></br></br></br><a class="linkedIn" href=""><content >LinkedIn</content></a></div>
 +
</div>
  
−
                <p>The <b>sequence</b> we designed contains two <b>RIP (RNAIII Inhibiting Peptide)</b> sequences fused to two different export signal peptides for <i>E. coli</i> Type II Secretion System: <b>DsbA</b>  and <b>MalE</b>, placed on N-terminal. (image: Figure 1. Schematic representation of the RIP production cassette. The cassette is composed of RIP sequence (blue) fused to DsbA signal (green) and further RIP sequence again (green) fused to MalE signal (red).)<br><br></p>
+
<div class="block full">
−
            <div class="block two-third center">
+
−
            <img src="https://static.igem.org/mediawiki/2018/f/fd/T--Pasteur_Paris--BBa_K2616001.png">
+
−
            <div class="legend"><b>Figure 11: </b>proNGF and TEV production cassette </div>
+
−
              </div>
+
−
                <p>Once we received the sequence encoding for this production cassette named Seq8 (461bp) in commercial plasmid pEX-A258 by gene synthesis. Plasmids was amplified in competent <i>E. coli</i> DH5alpha. After bacteria culture and plasmid DNA extraction, we digested commercial vector with <b>EcoRI</b> and <b>PstI</b> restriction enzymes. We extracted the inserts from the gel and performed a ligation by using specific overlaps into <b>linearized pBR322</b> for RIP expression and into <b>pSB1C3</b> for iGEM sample submission.<br>
+
  
 +
</div>
 +
 +
  <!-- member-->
 +
  <div class="block title"> <h2>Léa Guerassimoff</h2> </div>
 +
    <div class="team-member">
 +
<div class="block half">
 +
    <img class="team-photo" src="https://static.igem.org/mediawiki/2018/b/b3/T--Pasteur_Paris--Team_Members_Lea.jpg"></br></br></br><a class="linkedIn" href="https://www.linkedin.com/in/léa-guerassimoff-252816162/?originalSubdomain=fr"><content >LinkedIn</content></a></div>
 +
<div class="block half">
 +
      <content>Léa is our little chemist from Chimie ParisTech who specializes in making everything she does, cute. Her kindness and good mood always delight everyone. She is a real ray of sunshine even when she blows a fuse on the Wiki. She is very organized and serious in what she undertakes, but don’t bother her if she is watching a video of a toy goat: toy goats make her crazy, like really crazy. Regardless, she is one of the best people I have met, thanks to iGEM. We are now bosom buddies, that’s why people nickname us Tic & Tac. </br><i> By Ellyn</i></content>
 +
</div>
 
     </div>
 
     </div>
−
            <div class="block half">
+
 
−
            <img src="https://static.igem.org/mediawiki/2018/4/46/T--Pasteur_Paris--PSB1C3_RIP.png">
+
<div class="block full">
−
                <div class="legend"><b>Figure 12: </b> Agarose 1% gel after electrophoresis of digested pSB1C3 containing Seq8 (Bba_K2616001) with PstI and EcoRI. All colonies except 1, 3 and 7 contained the insert. </div>
+
−
  </div>
+
−
            <div class="block half">
+
−
            <img src="https://static.igem.org/mediawiki/2018/9/95/T--Pasteur_Paris--_pBR322_RIP.png">
+
−
              <div class="legend"><b>Figure 13: </b> Agarose 1% gel after electrophoresis of digested pBR322 containing Seq8 (Bba_K2616001) with NdeI (lane 1 to 7) All colonies except colonies 2 and 7 contained the insert.  </div>
+
  
 
</div>
 
</div>
 +
 +
    <!-- member-->
 +
<div class="block title"> <h2>Samuel Jaoui</h2> </div>
 +
    <div class="team-member">
 +
<div class="block half">
 +
      <content>Samuel is a 20 year-old engineering student from Chimie ParisTech. Despite being the youngest member of our team, Samuel was the first person to imagine and believe in this ambitious project and convinced us to create NeuronArch. His skills in biological engineering and programming help us to model scientific concepts of our project. </br> <i>By Etienne</i></content>
 +
</div>
 +
<div class="block half">
 +
    <img class="team-photo" src="https://static.igem.org/mediawiki/2018/f/f1/T--Pasteur_Paris--Team_Members_Samuel.jpg"></br></br></br><a class="linkedIn" href="https://www.linkedin.com/in/samuel-jaoui-3009bb14b/"><content >LinkedIn</content></a></div>
 +
    </div>
 +
 
<div class="block full">
 
<div class="block full">
−
<p>We repeated the procedure (transformation in <i>E. coli</i> Stellar competent cells, bacteria culture, plasmid DNA extraction, digestion) and we proved that our vectors contained the insert by electrophoresis (Figure 12,13).<br>
 
−
               
 
  
 
</div>
 
</div>
 +
 +
<!-- member-->
 +
<div class="block title"> <h2>Manon Madelenat</h2> </div>
 +
    <div class="team-member">
 +
<div class="block half">
 +
    <img class="team-photo" src="https://static.igem.org/mediawiki/2018/0/06/T--Pasteur_Paris--Team_Members_Manon.png"></br></br></br><a class="linkedIn" href="https://www.linkedin.com/in/manon-madelenat/"><content >LinkedIn</content></a></div>
 +
<div class="block half">
 +
      <content>Behind her strong character, Manon hides a very big heart! She is studying biotechnologies at Sup’Biotech Paris. She is in her last year of study and she actually works in health and production fields. Her next step is to work in neuroscience and understand neurodegenerative diseases! She is an essential member of the team and is the queen of mini and midi prep! Manon is truly as beautiful on the inside as she is on the outside, trust me! </br><i> By Léa</i></content>
 +
    </div></div>
 +
 
<div class="block full">
 
<div class="block full">
−
                <p>Alignment of <b>Sequencing</b> Results then confirmed that pSB1C3 contained Seq8, <a href="http://parts.igem.org/Part:BBa_K2616001"> Bba_K2616001 </a>. </p>
 
−
            </div>
 
−
            <div class="block two-third center">
 
−
                <img src="https://static.igem.org/mediawiki/2018/9/9f/T--Pasteur_Paris--SequencingRIP.PNG">
 
−
                <div class="legend"><b>Figure 14: </b> Alignment of sequencing results for BBa_K2616001. Sequencing perform in pSB1C3 plasmid and one primer was designed (FOR1) to cover the whole sequence. Image from Geneious. Pairwise % Identity: 100%.  </div>
 
−
            </div>
 
−
                <div class="block full">
 
−
                <p>Once checked, we cloned our construct into the <i>Escherichia coli</i> <b>BL21(DE3) pLysS</b> strain, a specific dedicated strain to produce high amounts of desired proteins under a T7 promoter. Bacteria were grown in 25 mL culture, and <b>protein expression</b> was induced with different IPTG concentration when bacteria have entered in a phase of exponential growth (approximately at 0.8 OD 600 nm) at 37°C. Pellet was sonicated and supernatant was kept<br>
 
−
                After two hours induction, we centrifuged and collect supernatant and pellet separately.<br><br></p></div>
 
  
 +
</div>
  
−
        <div class="block full">
+
    <!-- member-->
−
                <h4 style="text-align: left;">Fluorescence reading experiments</h4><br><br>
+
<div class="block title"> <h2>Jonathan Naccache</h2> </div>
−
                <p>Since RIP is only a seven-aminoacid peptide, we were not able to check its production by classic SDS-PAGE. Thus, we tried to check its expression by <b>observing its effect</b> on <i>Staphylococcus aureus</i> growth and adhesion. We grew a <i>S. aureus</i> strain expressing GFP (Green Fluorescent Protein), gently provided by Dr. Jean-Marc Ghigo on 96-well microtiter plates with different fractions of supernatant or pellet of our BL21(DE3) pLysS bacterial cultures containing BBa_K26160001.<br><br></p>
+
    <div class="team-member">
−
<p>After 48h or more incubation, we washed the plates in order to discard planktonic bacteria, and read fluorescence (excitation at 485 nm and measuring emission at 510 nm).<br><br></p>
+
<div class="block half">
−
          </div>
+
<content>Jon is really motivated and commited to the lab work! Studying biotechnologies at Sup’Biotech Paris for the past five years, he is very passionate about synthetic biology and would like to continue in this field of study by getting his Master of research next year. In the team, he spends a lot of time in the lab but also on illustrator to make amazing explanatory schemes!
 +
      </br> <i> By Manon </i></content>
 +
    </div>
 +
<div class="block half">
 +
    <img class="team-photo" src="https://static.igem.org/mediawiki/2018/0/07/T--Pasteur_Paris--Team_Members_Jonathan.jpg"></br></br></br><a class="linkedIn" href="https://www.linkedin.com/in/jonaccache/"><content >LinkedIn</content></a></div>
 +
     
 +
</div>
  
−
            <div class="block half">
+
<div class="block full">
−
                <img src="https://static.igem.org/mediawiki/2018/d/dd/T--Pasteur_Paris--FluorescenceResults1.png">
+
−
                <div class="legend"><b>Figure 15: </b>Measurement of GFP fluorescence from <i>S. aureus</i> biofilms cultivated with different IPTG induction concentrations of RIP peptide. Every measure was done eight times and the bars show the average fluorescence. CM= Culture Medium from the induced <i>E. coli</i> culture.. SL = Lysis Supernatant from the induced <i>E. coli</i> culture.</div>
+
−
            </div>
+
  
−
        <div class="block half">
+
</div>
−
                <p>Some of the results we got were extremely encouraging. For example, figure 5 shows an average 3-fold reduction of fluorescence from <i>S. aureus</i> biofilms when they were cultivated in presence of the bacterial lysate of an induced culture of BL-21 <i>E. coli</i> transformed with  BBa_K2616001. </p>
+
−
                <p>However, we performed experiments several times, and the results were not always as concluding. This variability is very likely due to a bias due regarding different approaches used for supernatant removal and washes. When using the flicking approach, we damaged our biofilm. Then, we removed planktonic cells by micropipette. </p>
+
−
            </div>
+
  
−
        <div class="block full">
 
−
                <h4 style="text-align: left;">Crystal violet staining</h4><br><br>
 
−
                <p>Since fluorescence measurements were not satisfying enough, we tried to improve our methods to quantify biofilm formation. Thus, we began to color biofilms by Crystal violet 0.1% staining and measuring absorbance at 570 nm. Again, the results were very heterogeneous between our different experiments, and between the different protocols. For instance, we tried to compare our protocol to WPI Worcester team's staining protocol, and the results, given in Figue 6 and 7 significantly differ.</p>
 
−
          </div>
 
  
−
            <div class="block half">
+
    <!-- member-->
−
                <img src="https://static.igem.org/mediawiki/2018/f/fa/T--Pasteur_Paris--CVPasteur.png">
+
  <div class="block title"> <h2>Éléa Paillares</h2> </div>
−
            </div>
+
    <div class="team-member">
 +
<div class="block half">
 +
    <img class="team-photo" src="https://static.igem.org/mediawiki/2018/8/86/T--Pasteur_Paris--Team_Members_Elea.jpg"></br></br></br><a class="linkedIn" href="https://www.linkedin.com/in/el%C3%A9a-paillares-b3305911a/"><content >LinkedIn</content></a></div>
 +
<div class="block half">
 +
      <content>Motivated and always smiling, Éléa always sets a happy mood in the room and never misses an occasion to crack you up. Although she spends her days locked in the laboratory working hard, making use of the skills she learned at Sup’Biotech, she has the spirit of a true traveler. Sometimes, she likes to go on spontaneous trips, not even bothering to book a hotel beforehand. She looks forward to the Giant Jamboree: she has never been to the United States!
 +
    </br> <i>By Andreas </i></content>
 +
    </div> </div>
  
−
            <div class="block half">
+
<div class="block full">
−
                <img src="https://static.igem.org/mediawiki/2018/9/9f/T--Pasteur_Paris--CVWPI.png">
+
−
            </div>
+
−
+
−
<div class="block full">
+
−
<div class="legend"><b>Figure 16: </b>Measurement of absorbance at 570 nm <i>S. aureus</i> biofilms cultivated with different IPTG induction concentrations of RIP peptide and stained with Crystal violet. Every measure was done eight times and the bars show the average fluorescence. CM= Culture Medium from the induced <i>E. coli</i> culture.. SL = Lysis Supernatant from the induced <i>E. coli</i> culture.</div>
+
−
            </div>
+
  
 +
</div>
  
−
        <div class="block two-third">
+
    <!-- member--> 
−
                <h4 style="text-align: left;">Biofilm PFA fixation before staining</h4><br><br>
+
<div class="block title"> <h2>Emma Picaud-Lucet</h2> </div>
−
<p>We wanted to avoid biofilm damage or loss during theses steps. In order to do that, we used Bouin solution to fix the formed biofilm after 24 and 48 hours of culture. Then biofilms were either stained with Crystal Violet 0.1% and resuspended in acetic acid 30% or resuspended in PBS 1X.  Surprisingly, with this method biofilm formation was higher when cultivated with cell extracts containing RIP. A that time, we are not able to explain why.</p>
+
    <div class="team-member">
 +
<div class="block half">
 +
      <content> What is more important to Emma than her cat’s happiness? IGEM of course! As a law student from the Faculté Jean Monnet, she is one of the members in charge of all of the legal aspects of the project. Never afraid of giving her opinions on various matters, she often deals with subjects beyond law, such as the human practices collaboration with other iGEM teams. Her secret talent: she’s a very talented cartoonist! </br><i> By Samuel </i></content>
 +
    </div>
 +
<div class="block half">
 +
 
 +
      <img class="team-photo" src="https://static.igem.org/mediawiki/2018/e/e8/T--Pasteur_Paris--Team_Members_Emma.jpg"></br></br></br><a class="linkedIn" href=""><content >LinkedIn</content></a></div>
 
</div>
 
</div>
  
−
            <div class="block one-third">
+
<div class="block full">
−
                <img src="https://static.igem.org/mediawiki/2018/f/f1/T--Pasteur_Paris--96-culture-wells-2.jpg">
+
−
                <div class="legend"><b>Figure 17: </b>Biofilm culture fixed with Bouin's solution in 96-well micrometer plate</div>
+
−
            </div>
+
  
 +
</div>
 +
 +
    <!-- member-->
 +
<div class="block title"> <h2>Sarah Porte</h2> </div>
 +
<div class="team-member">
 +
<div class="block half">
 +
    <img class="team-photo" src="https://static.igem.org/mediawiki/2018/6/64/T--Pasteur_Paris--Team_Members_Sarah.jpg"></br></br></br><a class="linkedIn" href="https://fr.linkedin.com/in/sporte"><content >LinkedIn</content></a></div>
 +
<div class="block half">
 +
      <content>Sarah is a biology major getting her Master’s at the sorbonne-Université Pierre et Marie Curie. She is highly organized and is very good at multi-tasking, which is very useful. Sarah appreciates her independence as she works best this way, but will lend a helping hand if she is needed. She will also gladly borrow other people’s sweaters if it helps her stay warm! </br>
 +
      <i> By Deshmukh </i></content>
 +
    </div> </div>
  
 
<div class="block full">
 
<div class="block full">
−
<p>With more time, we would certainly have been able to optimize our protocols to best fit with the strain we use, but for the time being, we are not able to give a final conclusion on whether or not our RIP peptide inhibits <i>S. aureus</i> biofilm formation.
 
−
<br><br></p></div>
 
  
−
            <div class="block full">
 
−
            <h2><i>S. aureus</i></b> Detection and RIP secretion <a href="http://parts.igem.org/Part:BBa_K2616003"> BBa_K2616003</a></h2><br><br>
 
−
<p>The sequence we designed contains the <i> agr </i> detection system from <i>S. aureus</i> and secretion of  RIP (RNAIII Inhibiting Peptide) sequences fused to two different export signal peptides for <i>E. coli</i> Type II Secretion System: DsbA and MalE, placed in N-terminal.</p>
 
 
</div>
 
</div>
  
−
            <div class="block two-third center">
+
    <!-- member-->
−
                <img src="https://static.igem.org/mediawiki/2018/7/77/T--Pasteur_Paris--ImproveParts.png">
+
<div class="block title"> <h2>Ellyn Redheuil</h2> </div>
−
                <div class="legend"><b>Figure 18: </b> <I> S. aureus </I> sensor device and RIP production cassette</div>
+
<div class="team-member">
−
            </div>
+
<div class="block half">
 +
      <content>Nicknamed Tic because of her inseparable bond with Léa (aka Tac), Ellyn lit up our summer with her continuous smiles, laughter, and amazing ideas and animations for our wiki page! She is an engineer student at ESPCI Paris, and you would be impressed by the range of her abilities: from coding the wiki to transforming bacteria. Whether in the lab or behind her computer, she seems to know how to do anything and everything. Her best specialty is her delicious cookies which are better than anyone else’s! </br><i> By Charlotte </i></content>
 +
    </div>
 +
<div class="block half">
 +
    <img class="team-photo" src="https://static.igem.org/mediawiki/2018/b/b1/T--Pasteur_Paris--Team_Members_Ellyn.jpg"></br></br></br><a class="linkedIn" href="https://www.linkedin.com/in/ellyn-redheuil-64a521b0/"><content >LinkedIn</content></a></div></div>
  
 
<div class="block full">
 
<div class="block full">
−
<p>Once we received the sequence encoding for this production cassette, named Seq5 (1422 bp), Seq6 (960 bp) and Seq7 (762 bp) in commercial plasmid pEX-A258 by gene synthesis.  Plasmids was amplified in competent <i>E. coli</i> DH5alpha. <br><br>
 
−
After bacterial culture and plasmid DNA extraction, we digested the commercial vector with XbaI and BamHI for Seq5, MscI and SphI for Seq6, HindII and SpeI for Seq7 restriction enzymes. We extracted the insert from the gel and ligated by specific overlaps into linearized pBR322 for expression and into pSB1C3 for iGEM sample submission.</p>
 
  
−
<p>We had trouble to proceed the ligation of the three inserts to linearized pBR322 and pSB1C3. We discussed with Takara Bio about our ligation issues, the GC percentage on our overlaps was to high to allow a good ligation.  Due to the lack of time we were not able to re design the overlaps for this construction.  </p>
+
</div>
−
            </div>
+
  
−
            <div class="block separator-mark"></div>
+
    <!-- member-->
−
                    </div>
+
<div class="block title"> <h2>Charlotte Richard</h2> </div>
 +
    <div class="team-member">
 +
<div class="block half">
 +
    <img class="team-photo" src="https://static.igem.org/mediawiki/2018/8/82/T--Pasteur_Paris--Team_Members_Charlotte.jpg"></br></br></br><a class="linkedIn" href=""><content >LinkedIn</content></a></div>
 +
<div class="block half">
 +
      <content>Engineer, biologist, adventurer and athlete, Charlotte has it all ! She is in her second year at Ecole Polytechnique, a military engineer school, and wants to specialize in Biology. In the lab, she is the best at Mini-preps, but she is also involved in the modeling part. In her spare time, she likes scouting and teaching the youth how to survive in the wild. That’s why she has the best BBQ skills!
 +
      </br> <i> By Gabriela </i> </content>
 +
    </div> </div>
  
−
        <div class="block full" style="background-color: transparent;">
+
<div class="block full">
−
                <i><p>Achievements:<br></i>
+
−
                    <ul style="text-align: left;">
+
−
                        <li>Successfully cloned a part coding for RIP secretion in pBR322 and in pSB1C3, creating a new part <a href="http://parts.igem.org/Part:BBa_K2616001"> Bba_K2616001 </a>.
+
−
                        <li>Successfully sequenced <a href="http://parts.igem.org/Part:BBa_K2616001"> Bba_K2616001 </a> in pSB1C3 and sent to iGEM registry.
+
−
                        <li>Successfully cultivated S. aureus biofilms in 96 well plates with different supernatants.</li>
+
−
                    </ul><br></p>
+
−
                    <p><i>Next steps:<br></i>
+
−
                    <ul style="text-align: left;">
+
−
                        <li>Clone the sensor device with inducible RIP production upon S. aureus detection.</li>
+
−
                        <li>Improve the characterization of RIP effect on biofilm formation.</li>
+
−
                    </ul>
+
−
                </p>
+
  
−
      </div>
+
</div>
−
      </div>                   
+
  
−
<!-- Third Onglet Kill switch-->
+
    <!-- member-->
−
                <div class="block full bothContent">
+
  <div class="block title"> <h2>Gabriela Sachet</h2> </div>
−
                    <div class="block dropDown" id="Kill">
+
    <div class="team-member">
−
                        <h4>KILL SWITCH: Click to see more </h4>
+
<div class="block half">
−
                    </div>
+
      <content>Studying biology at Sup’Biotech, Gabriela has played a key part in the laboratory. She has also contributed a lot to the bibliography, which helped keep the project on track. She has had the privilege of living in many different countries, which has made her a modern adventurer, always seeking new experiences. She recently participated in the 4L Rally (France to Morocco in a French retro car) before focusing on the iGEM competition !
 +
      </br> <i> By Claire </i> </content>
 +
    </div>
 +
<div class="block half">
 +
    <img class="team-photo" src="https://static.igem.org/mediawiki/2018/d/d8/T--Pasteur_Paris--Team_Members_Gabriela.jpg"></br></br></br><a class="linkedIn" href="https://www.linkedin.com/in/gabriela-sachet-570891b5/"><content >LinkedIn</content></a></div></div>
 +
 
 +
<div class="block full">
  
−
                    <div class="block hiddenContent">
+
</div>
−
                        <span class="closeCross"><img src="https://static.igem.org/mediawiki/2018/6/67/T--Pasteur_Paris--CloseCross.svg"></span>
+
 
−
                        <div class="block title">
+
<!-- member-->
−
                            <h1 style="padding-top: 50px;">KILL SWITCH</h1>
+
<div class="block title"> <h2>Thomas Starck</h2> </div>
 +
    <div class="team-member">
 +
<div class="block half">
 +
    <img class="team-photo" src="https://static.igem.org/mediawiki/2018/e/e5/T--Pasteur_Paris--Team_Members_Thomas.jpg"></br></br></br><a class="linkedIn" href="https://www.linkedin.com/in/thomas-starck-8b3828148/"><content >LinkedIn</content></a></div>
 +
<div class="block half">
 +
      <content>Spirit-son of Tony Stark, great defender of planet Earth and member of the Avengers, Thomas Starck chose a different path. Full of imagination and a practical spirit, Thomas is in his second year in Ecole Polytechnique, a military engineer school. In addition to his lab work, he also dedicated his time to the making of this wiki page. In his free time, he loves to help his “dad” save the world, but also go out and enjoy life.
 +
      </br><i> By Sarah </i></content>
 +
    </div> </div>
 +
 
 +
<div class="block full">
  
−
      </div>
 
−
      <div class="block half">
 
−
        <p>Once we received the sequences encoding for this production cassette (named construction Seq9) in commercial plasmids, in order to have more DNA, we transformed competent bacteria <i>E. coli</i> DH5alpha resulting in clones. After bacteria culture and plasmid DNA extraction, we digested commercial vectors with restriction enzymes, extracted the inserts from the gel, and ligated it into linearized pSB1C3 for iGEM submission and expression in BL21(DE)3.</p>
 
−
                <p>We repeated the procedure (transformation in <i>E. coli</i> Stellar competent cells, bacteria culture, plasmid DNA extraction, digestion) and we proved that our vector contained the insert by electrophoresis.</p>
 
−
      </div>
 
−
            <div class="block half">
 
−
        <img src="https://static.igem.org/mediawiki/2018/c/c5/T--Pasteur_Paris--GelKS.png">
 
−
        <div class="legend"><b>Figure 7: </b> Agar gel after electrophoresis of digested pSB1C3 containing Seq9 (Bba_K2616002) in columns 6 to 11. Colonies 2 and 6 have the correct plasmid. </div>
 
 
</div>
 
</div>
  
 +
  <!-- member-->
 +
<div class="block title"> <h2>Florian Thomas</h2> </div>
 +
    <div class="team-member">
 +
<div class="block half">
 +
      <content>Florian has many skills in his designer toolbox, he studied cabinet-making and he is now finishing his studies at ENSCi les Ateliers. He enjoys the beauty of raw material and well made products, which is reflected in his work. While being a calm and thoughtful person, Florian still knows how to make his point, especially when it comes to the topic of the use of animals in research. Do not let his backwards cap and youthful smile fool you, Florian is actually our oldest team member! </br> <i> By Éléa </i> </content>
 +
  </br>
 +
    </div>
 +
<div class="block half">
 +
    <img class="team-photo" src="https://static.igem.org/mediawiki/2018/2/23/T--Pasteur_Paris--Team_Members_Florian.jpg"></br></br></br><a class="linkedIn" href="https://www.linkedin.com/in/florian-thomas-80b401aa/"><content >LinkedIn</content></a></div></div>
 +
 
<div class="block full">
 
<div class="block full">
−
                <p>Alignment of <b>Sequencing</b> Results then confirmed that pSB1C3 contained Seq9, <a href="http://parts.igem.org/Part:BBa_K2616002"style="font-weight: bold ; color:#85196a;"target="_blank"> Bba_K2616002 </a>. </p>
+
 
−
            </div>
+
</div>
−
            <div class="block two-third center">
+
 
−
                <img src="https://static.igem.org/mediawiki/2018/d/d1/T--Pasteur_Paris--Sequencing-KS.PNG">
+
    <!-- member-->
−
                <div class="legend"><b>Figure 21: </b> Alignment of sequencing results for BBa_K2616002. Sequencing perform in pSB1C3 and two primers were designed (FOR1 and FOR2) to cover the whole sequence. Image from Geneious. Pairwise Identity: 96.9%.  </div>
+
<div class="block title"> <h2>Kelly Trang</h2> </div>
−
            </div>
+
    <div class="team-member">
 +
<div class="block half">
 +
    <img class="team-photo" src="https://static.igem.org/mediawiki/2018/f/f7/T--Pasteur_Paris--Team_Members_Kelly.jpg"></br></br></br><a class="linkedIn" href="https://www.linkedin.com/in/kelly-trang-134558123/"><content >LinkedIn</content></a></div>
 +
<div class="block half">
 +
      <content><p>Don’t let her fool you, behind her cute smile and freckles Kelly is actually a big fan of hardcore heavy metal. A first year Master’s student studying intellectual property law at the Faculté Jean Monnet, she is also a true scientist at heart and loves to follow the scientists around in the lab. When she is not baking something delicious or stealing food from her team members, she is very responsible and in charge of the legal work.</p> </br> <i> By Antoine </i></content>
 +
</content></a>
 +
    </div></div>
 +
 
 +
 
 +
 
 +
 
 +
</section>
 +
 
 +
 
 +
<section class="cf team-container">
 +
  <h1 class="team-h1">Supervisors</h1>
 +
 
 +
 
 +
  <!-- member-->
 +
<div class="block title" id="Deshmukh"> <h2>Deshmukh Gopaul</h2> </div>
 +
    <div class="team-member">
 +
<div class="block half">
 +
    <img class="team-photo" src="https://static.igem.org/mediawiki/2018/c/c0/T--Pasteur_Paris--Team_Members_Deshmukh.jpg"></br></br></br><a class="linkedIn" href=""><content >LinkedIn</content></a></div>
 +
<div class="block half">
 +
      <content>Deshmukh Gopaul, researcher and Head of Design in Biology at the Institut Pasteur, has been coaching the iGEM Pasteur Paris teams for 4 years now. Although he specializes in Biology, he is well versed in a variety of topics, allowing him to always have an interesting anecdote to share. He is very meticulous and although working with him may take a lot of time, patience, and tears, we know that it’s worth it in the end! </br><i> By Kelly </i></content>
 +
    </div></div>
  
 
<div class="block full">
 
<div class="block full">
−
                <p>The construction was successfully assembled. On Figure 21, mismatches are visible which correspond to the reduced precision of sequencing after 600 bp. To avoid this lack of precision, we used two different primers, allowing us to cover the whole sequence without mistakes. As visible, the mismatches are only present at the extremities of each primer sequencing. </p>
 
−
            </div>
 
  
 +
</div>
  
−
            <div class="block full">
+
    <!-- member-->
−
        <p>To test the efficiency of our kill-switch, we decided to cultivate BL21(DE)3 E. coli transformed with it at several temperatures (15°C, 20°C, 25°C and 37°C). The growth was followed by measuring the optical density at 600nm every 30 minutes for 6 hours, followed by two additional points at 18 hours and at 72 hours. Each experiment was done in a triplicate and the standard deviations were calculated for every point.  We show that the bacteria transformed with the kill-switch showed <b>no measurable growth</b> at 15°C and at 20°C during the 72 hours of the experiment, whereas the control population grew normally.</p>
+
<div class="block title" id="Guillian"> <h2>Guillian Graves</h2> </div>
−
                <p>At 25°C, the kill-switch population grew more slowly than the control for the first 18 hours, but the growth eventually started to reach normal values at 72 hours. </p>
+
    <div class="team-member">
−
                <p>Finally, at 37°C there was no difference in the growth of the kill-switch population compared to the control bacteria. </p>
+
<div class="block half">
−
      </div>
+
      <content> Guillian is an industrial designer specialized in biomimicry, working in Paris at his design studio BIG BANG Project. He also teaches at ENSCI-Les Ateliers (National School for Advanced Studies in Design). His professionalism and his high expectations go hand in hand with his sympathy. He helps our team think about the design of our device and uses his expertise to support our designers in creating a global project. </br> <i> By Claire </i></content>
−
            <div class="block two-third center">
+
−
        <img src="https://static.igem.org/mediawiki/2018/1/1b/T--Pasteur_Paris--kill-switch-graph-no-title.png">
+
−
        <div class="legend"><b>Figure 22: </b>Effect of different temperatures on the growth of Cryodeath kill-switch transformed BL21 <i>E. coli</i></div>
+
−
      </div>
+
−
            <div class="block full">
+
−
        <p>Thus, we successfully guarantee that our engineered bacteria will not be able to grow if they happened to be released in the environment.</p>
+
−
      </div>
+
−
            <div class="block separator-mark"></div>
+
 
     </div>
 
     </div>
−
        <div class="block full" style="background-color: transparent;">
+
<div class="block half">
−
<p><i><p>Achievements:<br></i>
+
      <img class="team-photo" src="https://static.igem.org/mediawiki/2018/b/be/T--Pasteur_Paris--Team_Members_Guillian.jpg"></br></br></br><a class="linkedIn" href=""><content >LinkedIn</content></a></div></div>
−
          <ul style="text-align: left;" style="text-align: left;">
+
−
            <li>Successfully cloned a part coding for toxin/antitoxin (CcdB/CcdA) system in  iGEM plasmid backbone, creating a <b>new composite part</b></li>
+
−
            <li>Successfully observe survival of our engineered bacteria at 25°C and 37°C and <b>absence of growth</b> at 18°C and 20°C, showing the <b>efficiency of the kill switch</b></li>
+
−
          </ul><br></p>
+
−
          <p><i>Next steps:</i><br>
+
−
          <ul style="text-align: left;">
+
−
            <li>Find a system that kills bacteria when released in the environment rather than just stopping their growth</li>
+
−
          </ul>
+
−
        </p></p></div>
+
−
    </div>  
+
  
 +
<div class="block full">
  
−
<!-- Fourth Onglet Membrane-->
+
</div>
−
                <div class="block full bothContent">
+
−
                    <div class="block dropDown" id="Membrane">
+
−
                        <h4>MEMBRANE: Click to see more</h4>
+
−
                    </div>
+
  
  
−
                    <div class="block hiddenContent">
+
    <!-- member-->
−
                        <span class="closeCross"><img src="https://static.igem.org/mediawiki/2018/6/67/T--Pasteur_Paris--CloseCross.svg"></span>
+
<div class="block title" id="Serena"> <h2>Serena Petracchini</h2> </div>
−
                        <div class="block title" id ="Membrane">
+
    <div class="team-member">
−
                            <h1 style="padding-top: 50px;">Membrane</h1>
+
  <div class="block half">
−
            </div>
+
    <img class="team-photo" src="https://static.igem.org/mediawiki/2018/b/b3/T--Pasteur_Paris--Team_Members_Serena.jpg"></br></br></br><a class="linkedIn" href=""><content >LinkedIn</content></a></div>
 +
<div class="block half">
 +
      <content>Serena has been the key to answering many of our question, thanks to her extensive experience in microbiology. Her major contribution to the project as a coach has been in the Interlab, along with the many great ideas she has! When she doesn’t agree with something you will hear her Italian come out as she says “ma no!” followed, of course, by some hand motion. If still you disagree with her, try her delicious tiramisu. I am sure she (it) will convince you! </br> <i> By Anna </i> </content>
 +
    </div> </div>
  
−
                    <div class="block two-third">
+
<div class="block full">
−
                        <p>The membrane filter is a key element of our prosthesis system, allowing the confinement of the genetically modified bacteria and the conduction of neuron impulses. We tested two types of membranes: Sterlitech Polycarbonate Gold-Coated Membrane Filters (pores diameter of 0.4 micrometer) and Sterlitech Alumina Oxide Membrane Filters (pores diameter of 0.2 micrometer).<br>
+
−
                        Sterlitech Alumina Oxide Membrane Filters were coated with different types of biocompatible conductive polymers: PEDOT:PSS (poly(3,4-ethylenedioxythiophene) polystyrene sulfonate), PEDOT:Cl and PEDOT:Ts.<br>
+
−
                        To characterize the potential of the different types of membranes to be integrated in our prosthesis system, we evaluated the coating of the alumina oxide membranes, their biocompatibility and their electrical conductivity.<br></p>
+
−
      </div>  
+
  
−
                    <div class="block one-third">
+
</div>
−
        <img src="https://static.igem.org/mediawiki/2018/9/9c/T--Pasteur_Paris--PEDOT-PSS.jpg">
+
−
        <div class="legend"><b>Figure 23: </b> PEDOT:PSS </div>
+
−
      </div>  
+
  
−
                    <div class="block full">
 
−
        <h2 style="text-align: left;">Biocompatibility</h2>
 
−
        <p></p>
 
−
      </div>
 
  
−
                    <div class="block full">
+
    <!-- member-->
−
        <h2 style="text-align: left;">Conductivity</h2>
+
    <div class="block title" id="Anna"> <h2>Anna Segú Cristina</h2> </div>
−
        <p></p>
+
    <div class="team-member">
−
      </div>  
+
<div class="block half">
 +
      <content>With a true catalan heart and a great french accent, Anna is putting the best of her energies in her tutoring role for iGem Pasteur. A soon-to-be second year PhD student, she finds time between a neuron an a miniprep to do sports and other extracurriculars, always leaving her friends astonished. Full of board game evenings and genetic engineering, she’s able to enjoy the best of Parisian life!  </br> <i>  By Serena </i> </content>
 +
    </div>
 +
<div class="block half">
 +
    <img class="team-photo" src="https://static.igem.org/mediawiki/2018/d/dd/T--Pasteur_Paris--Team_Members_Anna.jpg"></br></br></br><a class="linkedIn" href=""><content >LinkedIn</content></a></div></div>
  
−
                    <div class="block one-third">
+
<style>
−
        <img src="https://static.igem.org/mediawiki/2018/8/88/T--Pasteur_Paris--Well-chip.jpg" >
+
−
        <div class="legend"><b>Figure 24: </b> Hand-made PDMS well chip </div>
+
−
        </div> 
+
  
−
                    <div class="block two-third">
+
/* Team Member CSS
−
        <p>The conductivity of the membranes was measured on a self-made device. It consists of a culture well made of PDMS (polydimethylsiloxane), with a membrane filter at its bottom and a platinum wire linking the conductive membrane filter with the exterior.</p>
+
===========================*/
−
      </div>
+
  
−
                    <div class ="block full">
+
h1.team-h1 {
−
        <h3 style="text-align: left;">Platinum wire</h3>
+
  margin: 10;
−
        <p>The voltage difference between the two extremities of the wire was measured. </p>
+
  position:center;
−
      </div>  
+
  top: -16px;
 +
  text-align: center;
 +
  left: 40%;
 +
  font-family: raleway;
 +
  background-color: white;
 +
  padding: 0px 10px;
 +
  color: #000000;
 +
}
 +
 
 +
.cf:before,
 +
.cf:after {
 +
    content: " "; /* 1 */
 +
    display: table; /* 2 */
 +
}
 +
 
 +
.cf:after {
 +
    clear: both;
 +
}
 +
 
 +
/**
 +
* For IE 6/7 only
 +
* Include this rule to trigger hasLayout and contain floats.
 +
*/
 +
.cf {
 +
    *zoom: 1;
 +
}
 +
 
 +
.team-container {
 +
 
 +
  width: 100%;
 +
  border-top: 10px #e9e9e9 solid;
 +
  border-bottom: 10px #e9e9e9 solid;
 +
  padding-top: 5em;
 +
  padding-bottom: 5em;
 +
  margin-top: 3em;
 +
  position: relative;
 +
}
 +
 
 +
.team-member {
 +
  width:80%
 +
  float: left;
 +
  text-align: center;
 +
  margin-right: 0%;
 +
  margin-left: 0%
 +
}
 +
 
 +
.team-member:last-child {margin-right: 0;}
 +
 
 +
 
 +
.team-member content {
 +
  width: 30%;
 +
  font-family: raleway;
 +
  letter-spacing: -1px;
 +
  font-size: medium;
 +
 
 +
 
 +
}
 +
 
 +
.li {
 +
  color: #000000;
 +
 
 +
}
 +
 
 +
 
 +
 
 +
.team-photo {
 +
  border-radius: 50%;
 +
  text-align: center;
 +
  max-width: 60%;
 +
  height:auto;
 +
filter: opacity(50%);
 +
 +
  transition: 0.1s transform ease-in-out;
 +
}
 +
 
 +
 
 +
.team-photo:hover {
 +
  transform: scale(1.05);
 +
  filter: opacity(100%);
 +
}
 +
 
 +
 
 +
/* code for phone layout */
 +
@media (max-width:1000px){
 +
  .team-member {
 +
    width:25%;
 +
    margin-left: 4%;
 +
    margin-right: 4%;
 +
    margin-bottom: 40px;
 +
   
 +
  }
 +
 
 +
}
 +
 
 +
 
 +
 
 +
@media (max-width:1000px){
 +
  .team-member {
 +
    float: none;
 +
    display: block;
 +
    margin: 50px auto;
 +
    width: 100%;
 +
    text-align: center;
 +
  }
 +
 
 +
  .team-member content {
 +
    margin: 15px auto;
 +
  }
 +
 
 +
.team-photo {
 +
filter: opacity(100%);
 +
}
 +
 
 +
}
 +
 
 +
 
 +
/* ENDS Team Member CSS
 +
===========================*/
 +
 
 +
</style>
 +
 
 +
 
 +
 
 +
<!-- Inspired by
 +
MIT Licensed- Copyright (c) 2018 by Jose Morales-Mendizabal (https://codepen.io/moralejf/pen/YPxMpN)
 +
 
 +
 
 +
Permission is hereby granted, free of charge, to any person obtaining a copy of this software and associated documentation files (the "Software"), to deal in the Software without restriction, including without limitation the rights to use, copy, modify, merge, publish, distribute, sublicense, and/or sell copies of the Software, and to permit persons to whom the Software is furnished to do so, subject to the following conditions:
 +
 
 +
The above copyright notice and this permission notice shall be included in all copies or substantial portions of the Software.
 +
 
 +
THE SOFTWARE IS PROVIDED "AS IS", WITHOUT WARRANTY OF ANY KIND, EXPRESS OR IMPLIED, INCLUDING BUT NOT LIMITED TO THE WARRANTIES OF MERCHANTABILITY, FITNESS FOR A PARTICULAR PURPOSE AND NONINFRINGEMENT. IN NO EVENT SHALL THE AUTHORS OR COPYRIGHT HOLDERS BE LIABLE FOR ANY CLAIM, DAMAGES OR OTHER LIABILITY, WHETHER IN AN ACTION OF CONTRACT, TORT OR OTHERWISE, ARISING FROM, OUT OF OR IN CONNECTION WITH THE SOFTWARE OR THE USE OR OTHER DEALINGS IN THE SOFTWARE.
 +
-->
  
−
                    <div class="block two-third center">
 
−
        <img src="https://static.igem.org/mediawiki/2018/5/51/T--Pasteur_Paris--Wire-conductivity.PNG">
 
−
        <div class="legend"><b>Figure 25: </b> Voltage difference between the two extremities of the platinum wire.</div>
 
−
      </div>
 
  
−
                    <div class="block full">
 
−
        <p>The voltage difference between different platinum wires is pretty much the same. As we want to compare the differences between multiple membranes, we don't need to take into account the variability from one chip to another of the platinum wire's resistance. That means, it is meaningful to measure the voltage difference between a point on the membrane and the extremity of the wire outside the well, and use this data to compare the membranes. </p>
 
−
      </div>
 
  
−
                    <div class="block full">
 
−
        <h3 style="text-align: left;">Membranes</h3>
 
−
        <p>The voltage difference between a point on the membrane  (located near the border of the membrane filter) and the extremity of the platinum wire outside the well was measured.</p>
 
−
      </div>
 
  
−
                    <div class="block two-third center">
 
−
        <img src="https://static.igem.org/mediawiki/2018/2/2e/T--Pasteur_Paris--Membrane-conductivity.PNG">
 
−
        <div class="legend"><b>Figure 26: </b> Voltage difference between the extremity of the platinum wire outside the well and a point on the membrane.</div>
 
−
      </div>
 
  
  
−
      </div>
 
−
    </div>
 
−
  </div>
 
 
</html>
 
</html>

Latest revision as of 19:32, 16 October 2018

""

Let's meet our team!

Students

Etienne Axelos




LinkedIn
Etienne is one of our designers. He studies at the ENSCI-les Ateliers in Paris. He is always wearing black and fluorescent colors, which is very convenient because you can’t miss him. He is also a big music fan, and has special skills in rock’n’roll dance (very impressive!). As you can imagine, he is one of our funniest team members, you are never bored with him. But he is as funny as he is a hard worker, and we can always count on his help.
By Emma

Alice Dejoux

Calm, determined, attentive and engaged: that’s how we see Alice. This young Parisian born in Marseille is currently studying at the prestigious school Chimie Paristech. Talented and always smiling she was one of the leaders of the Parisian meet up organisation and she also attended the Mediterranean meet up in Marseille. Her scientific experience made her show great involvement and perseverance in the lab while doing the modeling of the project. She is part of the 23 strong links of the team iGEM Paris Pasteur 2018.
By Florian



LinkedIn

Claire Dumont




LinkedIn
Claire is one of our three marvelous designers from ENSCi les Ateliers. But that’s not it! She is also the key to keeping calm and focused the other two designers, who are more extroverted. Yet, she still loves to make movie references and silly jokes. In the team, she works very hard to create amazing graphics and she co-developed our final 3D-printed interface. It wouldn’t be an easy task to count the hours she spent on Rhino software to achieve her great results.
By Jonathan

Antoine Ehret

He is one of our dearest physicists studying at ESPCI Paris. He may seem like a shy person, but believe us: he is a man capable of wonders and knows a thing or two about inspirationnal speechs. In fact, you could consider him as our never ending source of encouragement. Each word that comes out of him fills you with determination. " You have to push back your limits each and everyday!"
By Aymen



LinkedIn

Andreas Fyrillas




LinkedIn
Behind his calm and hard-working look, Andreas Fyrillas will not hesitate to throw a punchline at you. His studies at the engineering school ESPCI Paris give him the skills to be an expert on microfluidic chips. Not only does he work the oscilloscope like no other, but he also has a secret talent in the kitchen that he uses to delight us from time to time. Apart from coding the wiki, he enjoys sharing his self-made dubstep music with us.
By Alice

Aymen Ghaneme

Aymen, currently a second-year student at CentraleSupelec, is a smiling Morrocan who is always in for a break and above all a great fruit sugar-free ice cream (in fact, after a while, we realized the break is only a pretext for the icecream). However, when he gets sufficient sleep, this hyper-active guy can use all his mathematical abilities on making models and will work for hours to solve any software bug.
By Thomas



LinkedIn

Léa Guerassimoff




LinkedIn
Léa is our little chemist from Chimie ParisTech who specializes in making everything she does, cute. Her kindness and good mood always delight everyone. She is a real ray of sunshine even when she blows a fuse on the Wiki. She is very organized and serious in what she undertakes, but don’t bother her if she is watching a video of a toy goat: toy goats make her crazy, like really crazy. Regardless, she is one of the best people I have met, thanks to iGEM. We are now bosom buddies, that’s why people nickname us Tic & Tac.
By Ellyn

Samuel Jaoui

Samuel is a 20 year-old engineering student from Chimie ParisTech. Despite being the youngest member of our team, Samuel was the first person to imagine and believe in this ambitious project and convinced us to create NeuronArch. His skills in biological engineering and programming help us to model scientific concepts of our project.
By Etienne



LinkedIn

Manon Madelenat




LinkedIn
Behind her strong character, Manon hides a very big heart! She is studying biotechnologies at Sup’Biotech Paris. She is in her last year of study and she actually works in health and production fields. Her next step is to work in neuroscience and understand neurodegenerative diseases! She is an essential member of the team and is the queen of mini and midi prep! Manon is truly as beautiful on the inside as she is on the outside, trust me!
By Léa

Jonathan Naccache

Jon is really motivated and commited to the lab work! Studying biotechnologies at Sup’Biotech Paris for the past five years, he is very passionate about synthetic biology and would like to continue in this field of study by getting his Master of research next year. In the team, he spends a lot of time in the lab but also on illustrator to make amazing explanatory schemes!
By Manon



LinkedIn

Éléa Paillares




LinkedIn
Motivated and always smiling, Éléa always sets a happy mood in the room and never misses an occasion to crack you up. Although she spends her days locked in the laboratory working hard, making use of the skills she learned at Sup’Biotech, she has the spirit of a true traveler. Sometimes, she likes to go on spontaneous trips, not even bothering to book a hotel beforehand. She looks forward to the Giant Jamboree: she has never been to the United States!
By Andreas

Emma Picaud-Lucet

What is more important to Emma than her cat’s happiness? IGEM of course! As a law student from the Faculté Jean Monnet, she is one of the members in charge of all of the legal aspects of the project. Never afraid of giving her opinions on various matters, she often deals with subjects beyond law, such as the human practices collaboration with other iGEM teams. Her secret talent: she’s a very talented cartoonist!
By Samuel



LinkedIn

Sarah Porte




LinkedIn
Sarah is a biology major getting her Master’s at the sorbonne-Université Pierre et Marie Curie. She is highly organized and is very good at multi-tasking, which is very useful. Sarah appreciates her independence as she works best this way, but will lend a helping hand if she is needed. She will also gladly borrow other people’s sweaters if it helps her stay warm!
By Deshmukh

Ellyn Redheuil

Nicknamed Tic because of her inseparable bond with Léa (aka Tac), Ellyn lit up our summer with her continuous smiles, laughter, and amazing ideas and animations for our wiki page! She is an engineer student at ESPCI Paris, and you would be impressed by the range of her abilities: from coding the wiki to transforming bacteria. Whether in the lab or behind her computer, she seems to know how to do anything and everything. Her best specialty is her delicious cookies which are better than anyone else’s!
By Charlotte



LinkedIn

Charlotte Richard




LinkedIn
Engineer, biologist, adventurer and athlete, Charlotte has it all ! She is in her second year at Ecole Polytechnique, a military engineer school, and wants to specialize in Biology. In the lab, she is the best at Mini-preps, but she is also involved in the modeling part. In her spare time, she likes scouting and teaching the youth how to survive in the wild. That’s why she has the best BBQ skills!
By Gabriela

Gabriela Sachet

Studying biology at Sup’Biotech, Gabriela has played a key part in the laboratory. She has also contributed a lot to the bibliography, which helped keep the project on track. She has had the privilege of living in many different countries, which has made her a modern adventurer, always seeking new experiences. She recently participated in the 4L Rally (France to Morocco in a French retro car) before focusing on the iGEM competition !
By Claire



LinkedIn

Thomas Starck




LinkedIn
Spirit-son of Tony Stark, great defender of planet Earth and member of the Avengers, Thomas Starck chose a different path. Full of imagination and a practical spirit, Thomas is in his second year in Ecole Polytechnique, a military engineer school. In addition to his lab work, he also dedicated his time to the making of this wiki page. In his free time, he loves to help his “dad” save the world, but also go out and enjoy life.
By Sarah

Florian Thomas

Florian has many skills in his designer toolbox, he studied cabinet-making and he is now finishing his studies at ENSCi les Ateliers. He enjoys the beauty of raw material and well made products, which is reflected in his work. While being a calm and thoughtful person, Florian still knows how to make his point, especially when it comes to the topic of the use of animals in research. Do not let his backwards cap and youthful smile fool you, Florian is actually our oldest team member!
By Éléa



LinkedIn

Kelly Trang




LinkedIn

Don’t let her fool you, behind her cute smile and freckles Kelly is actually a big fan of hardcore heavy metal. A first year Master’s student studying intellectual property law at the Faculté Jean Monnet, she is also a true scientist at heart and loves to follow the scientists around in the lab. When she is not baking something delicious or stealing food from her team members, she is very responsible and in charge of the legal work.


By Antoine

Supervisors

Deshmukh Gopaul




LinkedIn
Deshmukh Gopaul, researcher and Head of Design in Biology at the Institut Pasteur, has been coaching the iGEM Pasteur Paris teams for 4 years now. Although he specializes in Biology, he is well versed in a variety of topics, allowing him to always have an interesting anecdote to share. He is very meticulous and although working with him may take a lot of time, patience, and tears, we know that it’s worth it in the end!
By Kelly

Guillian Graves

Guillian is an industrial designer specialized in biomimicry, working in Paris at his design studio BIG BANG Project. He also teaches at ENSCI-Les Ateliers (National School for Advanced Studies in Design). His professionalism and his high expectations go hand in hand with his sympathy. He helps our team think about the design of our device and uses his expertise to support our designers in creating a global project.
By Claire



LinkedIn

Serena Petracchini




LinkedIn
Serena has been the key to answering many of our question, thanks to her extensive experience in microbiology. Her major contribution to the project as a coach has been in the Interlab, along with the many great ideas she has! When she doesn’t agree with something you will hear her Italian come out as she says “ma no!” followed, of course, by some hand motion. If still you disagree with her, try her delicious tiramisu. I am sure she (it) will convince you!
By Anna

Anna Segú Cristina

With a true catalan heart and a great french accent, Anna is putting the best of her energies in her tutoring role for iGem Pasteur. A soon-to-be second year PhD student, she finds time between a neuron an a miniprep to do sports and other extracurriculars, always leaving her friends astonished. Full of board game evenings and genetic engineering, she’s able to enjoy the best of Parisian life!
By Serena



LinkedIn