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<li><a href="https://2016.igem.org/Team:BGU_ISRAEL/Design">Design</a></li> | <li><a href="https://2016.igem.org/Team:BGU_ISRAEL/Design">Design</a></li> | ||
<li><a href="https://2016.igem.org/Team:BGU_ISRAEL/Experiments">Experiments</a></li> | <li><a href="https://2016.igem.org/Team:BGU_ISRAEL/Experiments">Experiments</a></li> | ||
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<li><a href="https://2016.igem.org/Team:BGU_ISRAEL/Results">Results</a></li> | <li><a href="https://2016.igem.org/Team:BGU_ISRAEL/Results">Results</a></li> | ||
<li><a href="https://2016.igem.org/Team:BGU_ISRAEL/Protocols">Protocols</a></li> | <li><a href="https://2016.igem.org/Team:BGU_ISRAEL/Protocols">Protocols</a></li> | ||
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<ul class="dropdown-menu submenubgu"> | <ul class="dropdown-menu submenubgu"> | ||
<li><a href="https://2016.igem.org/Team:BGU_ISRAEL/Parts#Overview">Overview</a></li> | <li><a href="https://2016.igem.org/Team:BGU_ISRAEL/Parts#Overview">Overview</a></li> | ||
− | <li><a href="https://2016.igem.org/Team:BGU_ISRAEL/Basic_Part">Basic | + | <li><a href="https://2016.igem.org/Team:BGU_ISRAEL/Basic_Part">Basic Parts</a></li> |
<li><a href="https://2016.igem.org/Team:BGU_ISRAEL/ImprovedParts">Improved Parts</a></li> | <li><a href="https://2016.igem.org/Team:BGU_ISRAEL/ImprovedParts">Improved Parts</a></li> | ||
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</ul> | </ul> | ||
</li> | </li> | ||
− | <li class="dropdown" id="humanB"><a class="dropdown-toggle" href="https://2016.igem.org/Team:BGU_ISRAEL/ | + | <li class="dropdown" id="humanB"><a class="dropdown-toggle" href="https://2016.igem.org/Team:BGU_ISRAEL/Integrated_Practices">Human Practices</a> |
<ul class="dropdown-menu submenubgu"> | <ul class="dropdown-menu submenubgu"> | ||
− | <li><a href="https://2016.igem.org/Team:BGU_ISRAEL/ | + | <li><a href="https://2016.igem.org/Team:BGU_ISRAEL/Integrated_Practices">Integrated Practices</a></li> |
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<li><a href="https://2016.igem.org/Team:BGU_ISRAEL/PlasticArt">Plastic Art</a></li> | <li><a href="https://2016.igem.org/Team:BGU_ISRAEL/PlasticArt">Plastic Art</a></li> | ||
− | <li><a href="https://2016.igem.org/Team:BGU_ISRAEL/ | + | <li><a href="https://2016.igem.org/Team:BGU_ISRAEL/EthicsAndSafety">Ethics & Safety</a></li> |
</ul> | </ul> | ||
</li> | </li> | ||
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<ul class="dropdown-menu submenubgu"> | <ul class="dropdown-menu submenubgu"> | ||
<li><a href="https://2016.igem.org/Team:BGU_ISRAEL/Entrepreneurship">Entrepreneurship</a></li> | <li><a href="https://2016.igem.org/Team:BGU_ISRAEL/Entrepreneurship">Entrepreneurship</a></li> | ||
− | <li><a href="https://2016.igem.org/Team:BGU_ISRAEL/ | + | <li><a href="https://2016.igem.org/Team:BGU_ISRAEL/Measurements">Measurements</a></li> |
+ | <li><a href="https://2016.igem.org/Team:BGU_ISRAEL/Proof">Proof Of Concept</a></li> | ||
+ | <li><a href="https://2016.igem.org/Team:BGU_ISRAEL/Demonstrate">Demonstrate</a></li> | ||
+ | <li><a href="https://2016.igem.org/Team:BGU_ISRAEL/HP/Silver">HP - Silver</a></li> | ||
+ | <li><a href="https://2016.igem.org/Team:BGU_ISRAEL/HP/Gold">HP - Gold</a></li> | ||
+ | <li><a href="https://2016.igem.org/Team:BGU_ISRAEL/Engagement">Engagements</a></li> | ||
</ul> | </ul> | ||
</li> | </li> | ||
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</nav> | </nav> | ||
− | <div class="container-fluid"> | + | <div class="container-fluid"> |
<div class="row"> | <div class="row"> | ||
<div class="PartColor"> | <div class="PartColor"> | ||
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<div class="container-fluid"> | <div class="container-fluid"> | ||
<div class='row whiteback'> | <div class='row whiteback'> | ||
− | + | <div class="col-lg-2"></div> | |
− | + | <div class="col-lg-8"> | |
− | + | <p class="bigAssHeader">Protocatechuate Degradation Pathway</p> | |
− | + | <hr> | |
− | + | <div class="row protoText"> | |
− | + | <div class="col-lg-6"> | |
− | + | <p> | |
− | + | <i>Pseudomonas putida</i> is a bacterium capable of utilizing protocatechuate, a toxic molecule for most bacteria. The protocatechuate degradation pathway converts protocatechuate into 3-oxoadipate, which is further metabolized by the bacteria and enters the TCA cycle. <br> | |
− | + | We chose to submit the genes encoding the enzymes in this pathway. | |
− | + | We also characterized the utilization of protocatechuate by bacterium using M9 minimal medium with protocatechuate as a sole carbon source. | |
− | + | We have not been able to submit the pcaB gene, encoding the 3-Carboxymuconate Cycloisomerase, since the sequence contained 4 PstI restriction cut sites, which we did not have time to alter using point mutations. | |
− | + | </p> | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
</div> | </div> | ||
− | <table class="table table-bordered"> | + | <div class="col-lg-6"> |
− | + | <img class="partPutipic" src="https://static.igem.org/mediawiki/2016/9/9e/Puti_biobricksBGU2016.png"> | |
− | + | </div> | |
− | + | </div> | |
− | + | <table class="table table-bordered"> | |
− | + | <thead> | |
− | + | <tr> | |
− | + | <th>Part Name</th> | |
− | + | <th>Type</th> | |
− | + | <th>Description</th> | |
− | + | <th>Designer</th> | |
− | + | <th>Length</th> | |
− | + | </tr> | |
− | + | </thead> | |
− | + | <tbody> | |
− | + | <tr> | |
− | + | <td><a href="http://parts.igem.org/Part:BBa_K2091000">BBa_K2091000</a></td> | |
− | + | <td>Coding</td> | |
− | + | <td>Protocatechuate 3,4-dioxygenase Alpha Subunit</td> | |
− | + | <td>B. Vaknin</td> | |
− | + | <td>606</td> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td><a href="http://parts.igem.org/Part:BBa_K2091001">BBa_K2091001</a></td> | |
− | + | <td>Coding</td> | |
− | + | <td>Protocatechuate 3,4-dioxygenase Beta Subunit</td> | |
− | + | <td>B. Vaknin</td> | |
− | + | <td>720</td> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td><a href="http://parts.igem.org/Part:BBa_K2091002">BBa_K2091002</a></td> | |
− | + | <td>Coding</td> | |
− | + | <td>4-Carboxymuconolactone Decarboxylase</td> | |
− | + | <td>B. Vaknin</td> | |
− | + | <td>393</td> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td><a href="http://parts.igem.org/Part:BBa_K2091003">BBa_K2091003</a></td> | |
− | + | <td>Coding</td> | |
− | + | <td>3-Oxoadipate enol-lactonase</td> | |
− | + | <td>B. Vaknin</td> | |
+ | <td>792</td> | ||
+ | </tr> | ||
+ | </tbody> | ||
+ | </table> | ||
</div> | </div> | ||
</div> | </div> | ||
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<div class="container-fluid"> | <div class="container-fluid"> | ||
<div class='row lightgold'> | <div class='row lightgold'> | ||
− | + | <div class="col-lg-2"></div> | |
− | + | <div class="col-lg-8"> | |
− | + | <p class="bigAssHeader">LC-Cutinase Protein Variants</p> | |
− | + | <hr> | |
− | + | <div class="row CutiText"> | |
− | + | <div class="col-lg-6"> | |
− | + | <p> | |
− | + | LC-cutinase is an enzyme found to have the ability to degrade PET. We have chosen to improve this enzyme using rational mutagenesis. 4 mutant variants of the LC-cutinase protein were produced using the PROSS algorithm and one Codon-optimized version of the protein was designed using the Genome Compiler software. <br> | |
− | + | We have tested and characterized these proteins using kinetic trials, bacterial growth assays and scanning electron microscopy imaging. <br> | |
− | + | </p> | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
</div> | </div> | ||
− | <table class='table table-bordered'> | + | <div class="col-lg-6"> |
− | + | <img class="partCutipic" src="https://static.igem.org/mediawiki/2016/0/00/Cuti_design_BGU2016.png"> | |
− | + | </div> | |
− | + | </div> | |
− | + | <table class='table table-bordered'> | |
− | + | <thead> | |
− | + | <tr> | |
− | + | <th>Part Name</th> | |
− | + | <th>Type</th> | |
− | + | <th>Description</th> | |
− | + | <th>Designer</th> | |
− | + | <th>Length</th> | |
− | + | </tr> | |
− | + | </thead> | |
− | + | <tbody> | |
− | + | <tr> | |
− | + | <td><a href="http://parts.igem.org/Part:BBa_K2091004">BBa_K2091004</a></td> | |
− | + | <td>Coding</td> | |
− | + | <td>LC-Cutinase Codon Optimized</td> | |
− | + | <td>I. Bariah, E. Zajfman, I. Segal</td> | |
− | + | <td>948</td> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td><a href="http://parts.igem.org/Part:BBa_K2091005">BBa_K2091005</a></td> | |
− | + | <td>Coding</td> | |
− | + | <td>LC-Cutinase Variant F4</td> | |
− | + | <td>I. Bariah, E. Zajfman, I. Segal</td> | |
− | + | <td>948</td> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td><a href="http://parts.igem.org/Part:BBa_K2091006">BBa_K2091006</a></td> | |
− | + | <td>Coding</td> | |
− | + | <td>LC-Cutinase Variant F7</td> | |
− | + | <td>I. Bariah, E. Zajfman, I. Segal</td> | |
− | + | <td>948</td> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td><a href="http://parts.igem.org/Part:BBa_K2091007">BBa_K2091007</a></td> | |
− | + | <td>Coding</td> | |
− | + | <td>LC-Cutinase Variant R4</td> | |
− | + | <td>I. Bariah, E. Zajfman, I. Segal</td> | |
− | + | <td>948</td> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td><a href="http://parts.igem.org/Part:BBa_K2091008">BBa_K2091008</a></td> | |
− | + | <td>Coding</td> | |
− | + | <td>LC-Cutinase Variant R7</td> | |
− | + | <td>I. Bariah, E. Zajfman, I. Segal</td> | |
+ | <td>948</td> | ||
+ | </tr> | ||
+ | </tbody> | ||
+ | </table> | ||
</div> | </div> | ||
</div> | </div> | ||
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}); | }); | ||
</script> | </script> | ||
+ | |||
<div class="container-fluid"> | <div class="container-fluid"> | ||
− | <div class="row"> | + | <div class="row footerColor"> |
− | < | + | <div class="col-lg-2"></div> |
+ | <div class="col-lg-4 text-center" > | ||
+ | <h4>Address:</h4> | ||
+ | <p> | ||
+ | Ben-Gurion University of the Negev<br> | ||
+ | Ben Gurion 1, Beer Sheva 8410501, Israel | ||
+ | </p> | ||
+ | <h5> | ||
+ | <b><u>Mail:</u></b> igembgu2016@gmail.com | ||
+ | </h5> | ||
+ | </div> | ||
+ | <div class="col-lg-4 text-center"> | ||
+ | <h4>Connect With Us!</h4> | ||
<div class="row"> | <div class="row"> | ||
− | <div class="col- | + | <div class="col-lg-3"></div> |
− | + | <div class="col-lg-6"> | |
− | + | <div class="row"> | |
− | + | <div class="col-lg-4 socialWrapper"> | |
− | + | <a href="https://www.facebook.com/iGEMBGU/?fref=ts" target="_blank"><img class="socialMediaIcon" src="https://static.igem.org/mediawiki/2016/7/7d/FacebookFlatIcon.png" alt="facebook"></a> | |
− | + | </div> | |
− | + | <div class="col-lg-4 socialWrapper"> | |
− | < | + | <a href=""><img class="socialMediaIcon" src="https://static.igem.org/mediawiki/2016/1/15/InstagramFlatIcon.png" alt="instagram"></a> |
− | + | </div> | |
− | + | <div class="col-lg-4 socialWrapper"> | |
− | + | <a href="https://twitter.com/igembgu2016" target="_blank"><img class="socialMediaIcon" src="https://static.igem.org/mediawiki/2016/3/34/TwitterFlatIcon.png" alt="twitter"></a> | |
− | + | </div> | |
− | + | </div> | |
− | + | ||
− | + | ||
− | + | ||
− | <a href="https://www.facebook.com/iGEMBGU/?fref=ts" target="_blank"><img class="socialMediaIcon" src=" | + | |
− | </ | + | |
− | < | + | |
− | <a ><img class="socialMediaIcon" src=" | + | |
− | </ | + | |
− | < | + | |
− | <a href="https://twitter.com/igembgu2016" target="_blank"><img class="socialMediaIcon" src=" | + | |
− | </ | + | |
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− | + | ||
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</div> | </div> | ||
</div> | </div> | ||
− | </ | + | </div> |
</div> | </div> | ||
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</html> | </html> |
Latest revision as of 14:55, 19 October 2016
Basic Part
Protocatechuate Degradation Pathway
Pseudomonas putida is a bacterium capable of utilizing protocatechuate, a toxic molecule for most bacteria. The protocatechuate degradation pathway converts protocatechuate into 3-oxoadipate, which is further metabolized by the bacteria and enters the TCA cycle.
We chose to submit the genes encoding the enzymes in this pathway.
We also characterized the utilization of protocatechuate by bacterium using M9 minimal medium with protocatechuate as a sole carbon source.
We have not been able to submit the pcaB gene, encoding the 3-Carboxymuconate Cycloisomerase, since the sequence contained 4 PstI restriction cut sites, which we did not have time to alter using point mutations.
Part Name | Type | Description | Designer | Length |
---|---|---|---|---|
BBa_K2091000 | Coding | Protocatechuate 3,4-dioxygenase Alpha Subunit | B. Vaknin | 606 |
BBa_K2091001 | Coding | Protocatechuate 3,4-dioxygenase Beta Subunit | B. Vaknin | 720 |
BBa_K2091002 | Coding | 4-Carboxymuconolactone Decarboxylase | B. Vaknin | 393 |
BBa_K2091003 | Coding | 3-Oxoadipate enol-lactonase | B. Vaknin | 792 |
LC-Cutinase Protein Variants
LC-cutinase is an enzyme found to have the ability to degrade PET. We have chosen to improve this enzyme using rational mutagenesis. 4 mutant variants of the LC-cutinase protein were produced using the PROSS algorithm and one Codon-optimized version of the protein was designed using the Genome Compiler software.
We have tested and characterized these proteins using kinetic trials, bacterial growth assays and scanning electron microscopy imaging.
Part Name | Type | Description | Designer | Length |
---|---|---|---|---|
BBa_K2091004 | Coding | LC-Cutinase Codon Optimized | I. Bariah, E. Zajfman, I. Segal | 948 |
BBa_K2091005 | Coding | LC-Cutinase Variant F4 | I. Bariah, E. Zajfman, I. Segal | 948 |
BBa_K2091006 | Coding | LC-Cutinase Variant F7 | I. Bariah, E. Zajfman, I. Segal | 948 |
BBa_K2091007 | Coding | LC-Cutinase Variant R4 | I. Bariah, E. Zajfman, I. Segal | 948 |
BBa_K2091008 | Coding | LC-Cutinase Variant R7 | I. Bariah, E. Zajfman, I. Segal | 948 |