Difference between revisions of "Team:TU-Eindhoven/Basic Part"

 
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                 <h1 style="text-align: center;">Parts overview</h1>
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                 <h1 style="text-align: center;">Basic Parts</h1>
 
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                            <p>The best that most of us can hope to achieve in physics is simply to misunderstand at a deeper level.</p>
 
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                     <p>To realize our designed parts, we gratefully made use of the IDT Synthesis offer,  
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                     <p>The list below shows our Basic Parts. For the medal requirements, we nominated <a href="http://parts.igem.org/Part:BBa_K2812000">BBa_K2812000</a> and <a href="http://parts.igem.org/Part:BBa_K2812004">BBa_K2812004</a> for the silver and gold medal, respectively. As new part for the silver medal, we created the carbohydrate binding domain, <a href="http://parts.igem.org/Part:BBa_K2812000">BBa_K2812000</a>, an adhesin binding strongly to glucose molecules. For the gold medal, TU-Eindhoven 2018 functionally improved on the original truncated lysostaphin <a href="http://parts.igem.org/Part:BBa_K748002">BBa_K748002</a> created by HIT-Harbin 2012 team in multiple ways. Through codon optimization, expression of the lysostaphin domain has been improved considerably. The other major functional improvement was realized by fusing the truncated lysostaphin to the HlyA secretion domain via a cleavable thrombin linker. In this way, (continuous) secretion via the Type I pathway can be achieved by co-expressing it with the HlyB/D secretion proteins. This avoids the requirement of cell lysis for release of the truncated lysostaphin, improving on the project of iGEM Harbin 2012, as self-destruction of their killer device is no longer necessary. Additionally, this new construct simplifies purification of truncated lysostaphin as running the medium over a nickel affinity column followed by thrombin cleavage results in pure truncated lysostaphin. The part <a href="http://parts.igem.org/Part:BBa_K2812004">BBa_K2812004</a> has been characterized extensively and the expression, secretion and selective activity of the truncated lysostaphin against S. aureus is demonstrated successfully.</p>
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                        allowing easy and fast access to the desired DNA constructs. All parts are codon
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                        optimized for expression in E. coli, the workhorse of our project. For all our composite parts,  
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                        the comprising basic parts were registered separately on the Registry, to allow future iGEM teams to
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                        use our constructs in new assemblies. All parts we shipped to the registry are sequence confirmed.
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                        We nominated <a href="http://parts.igem.org/Part:BBa_K2812004">BBa_K2812004</a>, lysostaphin-thrombin linker-HlyA-His tag, as Best New Basic Part as we think
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                        that the realisation of secretable lysostaphin opens up a whole new area of projects and applications,  
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                        since the issues and limitations associated with cell lysis are avoided. The T7-Lysostaphin-thrombin
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                        linker-HlyA-His part, <a href="http://parts.igem.org/Part:BBa_K2812005">BBa_K2812005</a>, has been nominated for Best New Composite Part, as it allows direct
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                        secretion of lysostaphin from the iGEM shipping backbone. This enables future teams to have a head start in
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                        their project.</p>
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                    <table class="parts-table">
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                      <caption>All Parts</caption>
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                      <tr>
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                        <th>Fave Part</th>
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                        <th>Name</th>
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                        <th>Type</th>
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                        <th>Description</th>
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                        <th>Designers</th>
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                        <th>Length</th>
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                      </tr>
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                      <tr class="odd-tr">
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                        <td><span style="color:red">❤</span></td>
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                        <td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2812000">BBa_K2812000</a></td>
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                        <td>Basic</td>
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                        <td>Carbohydrate-binding domain from MpIBP</td>
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                        <td>Guido Oerlemans, Maxime van den Oetelaar & Mariska Brüls</td>
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                        <td>576</td>
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                      </tr>
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                      <tr class="even-tr">
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                        <td></td>
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                        <td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2812001">BBa_K2812001</a></td>
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                        <td>Basic</td>
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                        <td>Coding sequence for trunctated Lysostaphin</td>
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                        <td>Guido Oerlemans, Maxime van den Oetelaar & Mariska Brüls</td>
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                        <td>738</td>
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                      </tr>
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                      <tr class="odd-tr">
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                        <td></td>
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                        <td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2812002">BBa_K2812002</a></td>
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                        <td>Basic</td>
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                        <td>pBAD-ara promoter - Arabinose inducible regulatory promoter </td>
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                        <td>Guido Oerlemans, Maxime van den Oetelaar and Mariska Brüls</td>
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                        <td>71</td>
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                      </tr>
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                      <tr  class="even-tr">
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                        <td></td>
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                        <td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2812003">BBa_K2812003</a></td>
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                        <td>Composite</td>
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                        <td>Coding sequence for trunctated Lysostaphin regulated by T7-promoter</td>
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                        <td>Guido Oerlemans, Maxime van den Oetelaar and Mariska Brüls</td>
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                        <td>776</td>
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                      </tr>
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                      <tr class="odd-tr">
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                        <td><span style="color:red">❤</span></td>
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                        <td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2812004">BBa_K2812004</a></td>
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                        <td>Basic</td>
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                        <td>Coding sequence for trunctated Lysostaphin fused to His-tagged HlyA</td>
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                        <td>Guido Oerlemans, Maxime van den Oetelaar & Mariska Brüls</td>
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                        <td>1458</td>
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                      </tr>
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                        <td><span style="color:red">❤</span></td>
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                        <td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2812005">BBa_K2812005</a></td>
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                        <td>Composite</td>
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                        <td>Coding sequence for trunctated Lysostaphin with HlyA and His6-tag regulated by T7-promoter</td>
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                        <td>Guido Oerlemans, Maxime van den Oetelaar & Mariska Brüls</td>
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                        <td>1496</td>
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                      </tr>
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                      <tr class="odd-tr">
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                        <td></td>
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                        <td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2812006">BBa_K2812006</a></td>
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                        <td>Basic</td>
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                        <td>Coding sequence for Pyocin S5 with HlyA and His6-tag </td>
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                        <td>Guido Oerlemans, Maxime van den Oetelaar & Mariska Brüls</td>
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                        <td>2217</td>
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                      </tr>
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                      <tr  class="even-tr">
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                        <td></td>
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                        <td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2812007">BBa_K2812007</a></td>
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                        <td>Composite</td>
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                        <td>Coding sequence for Pyosin S5 with HlyA and His6-tag regulated by AraC promoter</td>
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                        <td>Guido Oerlemans, Maxime van den Oetelaar & Mariska Brüls</td>
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                        <td>2288</td>
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                      </tr>
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                      <caption>All Parts</caption>
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                        <th>Fave Part</th>
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                        <th>Name</th>
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                        <th>Type</th>
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                        <td><span style="color:red">❤</span></td>
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                        <td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2812005">BBa_K2812005</a></td>
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                        <td>Composite</td>
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                        <td>Coding sequence for trunctated Lysostaphin with HlyA and His6-tag regulated by T7-promoter</td>
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−
                        <td>Guido Oerlemans, Maxime van den Oetelaar & Mariska Brüls</td>
+
−
                        <td>1496</td>
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                      </tr>
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                      <tr  class="odd-tr">
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                        <td></td>
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                        <td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2812003">BBa_K2812003</a></td>
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                        <td>Composite</td>
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−
                        <td>Coding sequence for trunctated Lysostaphin regulated by T7-promoter</td>
+
−
                        <td>Guido Oerlemans, Maxime van den Oetelaar and Mariska Brüls</td>
+
−
                        <td>776</td>
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−
                      </tr>
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−
 
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                      <tr  class="even-tr">
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                        <td></td>
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                        <td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2812007">BBa_K2812007</a></td>
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−
                        <td>Composite</td>
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−
                        <td>Coding sequence for Pyosin S5 with HlyA and His6-tag regulated by AraC promoter</td>
+
−
                        <td>Guido Oerlemans, Maxime van den Oetelaar & Mariska Brüls</td>
+
−
                        <td>2288</td>
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−
                      </tr>
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                    </table>
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{{:Team:TU-Eindhoven/footer}}

Latest revision as of 02:58, 18 October 2018

Basic Parts

The list below shows our Basic Parts. For the medal requirements, we nominated BBa_K2812000 and BBa_K2812004 for the silver and gold medal, respectively. As new part for the silver medal, we created the carbohydrate binding domain, BBa_K2812000, an adhesin binding strongly to glucose molecules. For the gold medal, TU-Eindhoven 2018 functionally improved on the original truncated lysostaphin BBa_K748002 created by HIT-Harbin 2012 team in multiple ways. Through codon optimization, expression of the lysostaphin domain has been improved considerably. The other major functional improvement was realized by fusing the truncated lysostaphin to the HlyA secretion domain via a cleavable thrombin linker. In this way, (continuous) secretion via the Type I pathway can be achieved by co-expressing it with the HlyB/D secretion proteins. This avoids the requirement of cell lysis for release of the truncated lysostaphin, improving on the project of iGEM Harbin 2012, as self-destruction of their killer device is no longer necessary. Additionally, this new construct simplifies purification of truncated lysostaphin as running the medium over a nickel affinity column followed by thrombin cleavage results in pure truncated lysostaphin. The part BBa_K2812004 has been characterized extensively and the expression, secretion and selective activity of the truncated lysostaphin against S. aureus is demonstrated successfully.

Basic Parts
Fave Part Name Type Description Designers Length
❤ BBa_K2812004 Basic Coding sequence for trunctated Lysostaphin fused to His-tagged HlyA Guido Oerlemans, Maxime van den Oetelaar & Mariska Brüls 1458
❤ BBa_K2812000 Basic Carbohydrate-binding domain from MpIBP Guido Oerlemans, Maxime van den Oetelaar & Mariska Brüls 576
BBa_K2812001 Basic Coding sequence for trunctated Lysostaphin Guido Oerlemans, Maxime van den Oetelaar & Mariska Brüls 738
BBa_K2812002 Basic pBAD-ara promoter - Arabinose inducible regulatory promoter Guido Oerlemans, Maxime van den Oetelaar and Mariska Brüls 71
BBa_K2812006 Basic Coding sequence for Pyocin S5 with HlyA and His6-tag Guido Oerlemans, Maxime van den Oetelaar & Mariska Brüls 2217
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