Team:Trieste/data

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        <h1 id="h1_lf" class="main_tit"><div>Data</div></h1>
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<h1 id="h1_lf" class="main_tit"><div>Data</div></h1>
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            <h1 id="h1_rt" class="main_tit"><div>More</div></h1>
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<h1 id="h1_rt" class="main_tit"><div>More</div></h1>
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                <div class="box_contenuti">
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<h1>An overview on our system</h1>
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<h1>An overview of our system</h1>
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<h2></h2>
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<center><img src="https://static.igem.org/mediawiki/2012/a/a8/Progetto_copia.jpg" width="700px"/></center>
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<br/>
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<p>
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<center><img src="https://static.igem.org/mediawiki/2012/a/a8/Progetto_copia.jpg" width="700px"/></center>
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<strong>This is a diagram summarising our system. It consists of two parts: the production of Ab against NoroVirus and the gene guard.</strong><br/>
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<br/>
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Concerning the gene guard, CymR inhibits the T5 Cumate Operator Promoter avoiding the production of the T4 Holin (permeabilizes the cytoplasmatic membrane) and the LL 37 cathelicidin (human antimicrobial peptide able to disrupt the outer membrane).<br/>
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<strong>This is a diagram summarising our system. It consists of two parts: the production of Ab against NoroVirus and the gene guard.</strong><br/>
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When the cumate is given, it binds and inactivates the CymR, derepressing the T5 Cumate Operator Promoter and allowing the expression of T4 Holin, that permeabilizes the cytoplasmatic membrane, allowing the secretion of LL 37 that disrupt the outer membrane.<br/>
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Concerning the gene guard, CymR inhibits the T5 Cumate Operator Promoter avoiding the production of the T4 Holin (permeabilizes the cytoplasmatic membrane) and the LL 37 cathelicidin (human antimicrobial peptide able to disrupt the outer membrane).<br/>
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Moreover, the horizontal transfer from our Nissle into another bacteria is avoided thanks to our gene guard. The receiving bacteria do not produce the CymR repressor, so the T5 Cumate Operator Promoter is activated allowing T4 Holin and LL 37 expression. Bacteria die!
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When the cumate is given, it binds and inactivates the CymR, derepressing the T5 Cumate Operator Promoter and allowing the expression of T4 Holin, that permeabilizes the cytoplasmatic membrane, allowing the secretion of LL 37 that disrupt the outer membrane.<br/>
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</p>
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Moreover, the horizontal transfer from our Nissle into another bacteria is avoided thanks to our gene guard. The receiving bacteria do not produce the CymR repressor, so the T5 Cumate Operator Promoter is activated allowing T4 Holin and LL 37 expression. Bacteria die!
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<h2> Our favourite parts:</h2>
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<br/>
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<table>
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<br/>
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<tbody>
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<h2> Our favourite parts:</h2>
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<tr>
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                          <table>
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<td><img src="http://mmcpoker.net/images/cuori.gif" width="50px"/></td>  
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    <tbody>
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<td>
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<tr>
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<h3><a href="https://2012.igem.org/Team:Trieste/parts/1">BBa_K875001</a>, T5 cumate operator:</h3>  
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    <td><img src="http://mmcpoker.net/images/cuori.gif" width="50px"/></a></td>  
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We demonstrated that this promoter is very strong and strictly regulated. Thanks to its rapid responsiveness it's one of the first candidates for the toxin expression and production in case of necessity.
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    <td><h3>BBa_K875001, T5 cumate operator:</h3>  
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</td>
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<br/>
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</tr>
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We have demonstrated that it is a very strong and strictly regulated promoter. Thanks to its rapid reactivity it's one of the first candidate for the expression and production of toxin in case of necessity.<br/>
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<tr>
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<br/></td>
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<td><img src="http://mmcpoker.net/images/cuori.gif" width="50px"/></td>  
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                </tr>
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<td>
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<tr>
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<h3><a href="https://2012.igem.org/Team:Trieste/parts/3">BBa_K875003</a>, CymR:</h3>
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    <td><img src="http://mmcpoker.net/images/cuori.gif" width="50px"/></a></td>  
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We expressed succesfully the CymR. We also showed his stringency as repressor of the T5 Cumate Operator. Together, CymR and T5 Cumate Operator, create a safe and tightly regulated gene guard system, which keeps under control the bacterial growth.
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    <td><h3>BBa_K875003, CymR:</h3>
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</td>
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<br/>
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</tr>
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We have succesfully expressed the CymR. We have also showed his stringency as repressor of the T5 Cumate Operator that makes the all system a very useful mechanism for an high specificity expression.
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<tr>
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<br/><br/></td>
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<td><img src="http://mmcpoker.net/images/cuori.gif" width="50px"/></td>  
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                </tr>
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<td>
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<tr>
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<h3><a href="ttp://2012.igem.org/Team:Trieste/parts/4">BBa_K875004</a>, LPP-OmpA-scFv:</h3>  
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    <td><img src="http://mmcpoker.net/images/cuori.gif" width="50px"/></a></td>  
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We verified the production of this protein at high yield. LPP-OmpA system is a very powerful method for protein display on the outer membrane, in this case the antibody scFv 54.6.
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    <td><h3>BBa_K875004, LPP-OmpA-scFv:</h3>  
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</td>
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<br/>
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</tr>
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We have verified the production of this protein at high yield. OmpA is a very powerful method for displaying proteins on the external membrane, in this case the antibody scFv 54.6.<br/><br/></td>
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</tbody>
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                </tr>
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</table>
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<tr>
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<h2>The part we improved:</h2>
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<br/>
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<table>
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<tbody>
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<br/></td>
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<tr>
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<td><img src="http://www.superscommesse.it/images/stella_piena.jpg" width="50px"/></a></td>  
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                            </tbody>
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<td>
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                          </table>
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<h3><a href="https://2012.igem.org/Team:Trieste/parts/10">BBa_K875020</a>, Beta-Glucosidase (Osaka 2010):</h3>  
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This composite part is developed by previous work by UNITS iGEM team 2011 and Edinburgh team 2011. We improved and characterized this part demonstrating its efficiency.
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</td>
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</tr>
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</tbody>
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</table>
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<h2>Other parts that we developed:</h2>
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<p>
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<h2>The part we have improved:</h2>
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<h3><a href="https://2012.igem.org/Team:Trieste/parts/2">BBa_K875002</a>, T5 Lac Operator:</h3>
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<table>
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We showed that this promoter comes very handy for protein expression tests.
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<tbody>
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</p>
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<tr>
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    <td><img src="http://www.superscommesse.it/images/stella_piena.jpg" width="50px"/></a></td>  
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<p>
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    <td><h3>BBa_K875020, Beta-Glucosidase (Osaka 2010):</h3>  
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<h3><a href="https://2012.igem.org/Team:Trieste/parts/5">BBa_K875005</a>, LPP-OmpA-SIP:</h3>
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<br/>
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We designed this biobrick containing SIP as a valid alternative to scFv, because SIP confers bivalent binding properties to the antibody.
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This composite part is developed by previous work by UNITS iGEM team 2011 and Edinburgh team 2011. We improved and characterized this part demonstrating its efficiency.<br/><br/></td>
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</p>
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                </tr>
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<p>
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</tbody>
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<h3><a href="https://2012.igem.org/Team:Trieste/parts/6">BBa_K875006</a>, PelB-scFv:</h3>
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</table>
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We created this composit biobrick to use as a system which allows secretion of proteins, in this case the antibody, first into periplasm and then extracellularly.
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</p>
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<br/>
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<p>
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<h3><a href="https://2012.igem.org/Team:Trieste/parts/7">BBa_K875007</a>, PelB-SIP:</h3>
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We built this biobrick (secreted version of SIP) because soluble form of SIP plays an important role in enteric infections.
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<h2>Other parts that we have developed:</h2>
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</p>
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<br/>
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<p>
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<h3>BBa_K875002, T5 Lac Operator:</h3>
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<h3><a href="https://2012.igem.org/Team:Trieste/parts/8">BBa_K875008</a>, Tse2 toxin:</h3>
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<br/>
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We demonstrated that this composit is an important suicide mechanism able to arrest the growth of prokaryotic cells.
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We have shown that this promoter is a very useful biobricks in the primary testing of protein expression thanks to his handiness.
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</p>
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<br/>
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<p>
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<br/>
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<h3><a href="https://2012.igem.org/Team:Trieste/parts/9">BBa_K875009</a>, LL 37:</h3>
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<h3>BBa_K875005, LPP-OmpA-SIP:</h3>
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The LL 37 is a very powerful human antimicrobial peptide that kills bacteria.
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<br/>
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We are developing a new construct combining the toxin with the T4 holin which breaks the inner membrane allowing the LL 37 to reach his target.
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We have built this biobrick as a valid alternative to the LPP-OmpA-scFv because it has an higher stability, even if its size is bigger than the scFv.
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</p>
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<br/>
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</div>
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</div> <!-- end box_main -->
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<h3>BBa_K875006, PelB-scFv:</h3>
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<div id="box_right"> <!-- start box_right -->
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<ul id="sub_menu">
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We have created this composit as an optimum system to allow the entry of proteins, in this case the antibody, in the bacterial periplasm and outside the outer mambrane according to the protein size.
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<li class="select"><a href="https://2012.igem.org/Team:Trieste/data">Data</a></li>
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</ul>
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<img src="https://static.igem.org/mediawiki/2012/b/b0/Team_trieste.jpg" alt="Team iGEM 2012" id="igem_team" />
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<h3>BBa_K875007, PelB-SIP:</h3>
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<div class="box_contacts">
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<br/>
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<h2>Contact us</h2>
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We have built this biobrick according to the trait of the SIP that acts an important role against the enteric infection when secreted.
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<p>For other information, write to:</p>
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<br/>
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<a href="mailto:igem2012@gmail.com" class="btn">igem2012@gmail.com</a>
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<br/>
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<div class="social">
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<h3>BBa_K875008, Tse2 toxin:</h3>
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Follow us also:
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<br/>
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<a href="https://www.facebook.com/IGEMUNITS?ref=ts" target="_blank"><img src="https://static.igem.org/mediawiki/2012/f/f4/Ico_fb.png" alt="Facebook" class="fb" /></a>
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We have demonstrated that this composit is a important mechanism through which we can eliminate bacteria from their intracellular space.<br/>
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<a href="https://twitter.com/iGEMTrieste" target="_blank"><img src="https://static.igem.org/mediawiki/2012/2/21/Ico_twitter.png" alt="twitter" class="tw" /></a>
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<h3>BBa_K875009, LL37-T4 holin:</h3>
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</div> <!-- end box_right -->
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We have projected this composit using  LL-37 which is a very powerful human antimicrobial peptide. LL-37 kills bacteria destroying the outer membrane, this toxin acts in combination with the T4 holin which breaks the inner membrane allowing the contact between LL-37 and the outer membrane.
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                <li class="select"><a href="https://2012.igem.org/Team:Trieste/data">Data</a></li>
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                        <li><a href="https://2012.igem.org/Team:Trieste/project/mainres">Main Results</a></li>
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<h2>Contact us</h2>
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<p>For other information, write to:</p>
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                    <a href="mailto:igem2012@gmail.com" class="btn">igem2012@gmail.com</a>
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                    <div class="social">
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                    Follow us also:
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<a href="https://www.facebook.com/IGEMUNITS?ref=ts" target="_blank"><img src="https://static.igem.org/mediawiki/2012/f/f4/Ico_fb.png" alt="Facebook" class="fb" /></a>
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                        <a href="https://twitter.com/igemunits" target="_blank"><img src="https://static.igem.org/mediawiki/2012/2/21/Ico_twitter.png" alt="twitter" class="tw" /></a>
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Latest revision as of 12:23, 15 October 2012

Data

More

An overview of our system

This is a diagram summarising our system. It consists of two parts: the production of Ab against NoroVirus and the gene guard.
Concerning the gene guard, CymR inhibits the T5 Cumate Operator Promoter avoiding the production of the T4 Holin (permeabilizes the cytoplasmatic membrane) and the LL 37 cathelicidin (human antimicrobial peptide able to disrupt the outer membrane).
When the cumate is given, it binds and inactivates the CymR, derepressing the T5 Cumate Operator Promoter and allowing the expression of T4 Holin, that permeabilizes the cytoplasmatic membrane, allowing the secretion of LL 37 that disrupt the outer membrane.
Moreover, the horizontal transfer from our Nissle into another bacteria is avoided thanks to our gene guard. The receiving bacteria do not produce the CymR repressor, so the T5 Cumate Operator Promoter is activated allowing T4 Holin and LL 37 expression. Bacteria die!

Our favourite parts:

BBa_K875001, T5 cumate operator:

We demonstrated that this promoter is very strong and strictly regulated. Thanks to its rapid responsiveness it's one of the first candidates for the toxin expression and production in case of necessity.

BBa_K875003, CymR:

We expressed succesfully the CymR. We also showed his stringency as repressor of the T5 Cumate Operator. Together, CymR and T5 Cumate Operator, create a safe and tightly regulated gene guard system, which keeps under control the bacterial growth.

BBa_K875004, LPP-OmpA-scFv:

We verified the production of this protein at high yield. LPP-OmpA system is a very powerful method for protein display on the outer membrane, in this case the antibody scFv 54.6.

The part we improved:

BBa_K875020, Beta-Glucosidase (Osaka 2010):

This composite part is developed by previous work by UNITS iGEM team 2011 and Edinburgh team 2011. We improved and characterized this part demonstrating its efficiency.

Other parts that we developed:

BBa_K875002, T5 Lac Operator:

We showed that this promoter comes very handy for protein expression tests.

BBa_K875005, LPP-OmpA-SIP:

We designed this biobrick containing SIP as a valid alternative to scFv, because SIP confers bivalent binding properties to the antibody.

BBa_K875006, PelB-scFv:

We created this composit biobrick to use as a system which allows secretion of proteins, in this case the antibody, first into periplasm and then extracellularly.

BBa_K875007, PelB-SIP:

We built this biobrick (secreted version of SIP) because soluble form of SIP plays an important role in enteric infections.

BBa_K875008, Tse2 toxin:

We demonstrated that this composit is an important suicide mechanism able to arrest the growth of prokaryotic cells.

BBa_K875009, LL 37:

The LL 37 is a very powerful human antimicrobial peptide that kills bacteria. We are developing a new construct combining the toxin with the T4 holin which breaks the inner membrane allowing the LL 37 to reach his target.

Team iGEM 2012

Contact us

For other information, write to:

igem2012@gmail.com
Università degli studi di Trieste ICGEB Illy Fondazione Cassa di Risparmio
iGEM 2012 iGEM 2012 iGEM 2012 iGEM 2012 iGEM 2012 iGEM 2012
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