Team:DTU-Denmark-2/Project/achievements

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<b>Achievements</b><br> <br>
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<b></b>Achievements <br>
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<a href="https://2011.igem.org/Team:DTU-Denmark-2/Project/achievements#Achievements" class="h1"> Achievements</a><br><br>
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<a href="https://2011.igem.org/Team:DTU-Denmark-2/Project/achievements#Fulfilment of medal requirements" class="h1"> Fulfilment of medal requirements</a><br><br>
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<a name="Achievements"></a><h1><b>Achievements</b></h1>
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The team came together during a course in mammalian cell biology in June, and that was when the idea of iGEM popped into our heads. Even though we started the iGEM project late June, we have managed to design a fully standardized cloning system called Plug'n Play with DNA. We thought that time should be spent on making cool iGEM projects, instead of struggling with unwanted restriction sites and cumbersome assembly systems. This led us to standardize USER fusion by designing the<a hret="https://2011.igem.org/Team:DTU-Denmark-2/Project/overview"> Plug'n'Play with DNA </a>assembly system.</p><br>
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<b>Our Plug 'n' Play with DNA kit consist of:</b><br>
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<li> 49 ready-to-use BioBricks.</li>
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To ensure that the BioBricks and devices have been assembled correctly we have tested the functionality of them in <i>Aspergillus nidulans</i> and in mammalian cells.
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We have characterized the two fungal promoters, P<i>alcA</i> and DMKP-P6,  and demonstrated function of the cytomegalovirus (CMV) promoter in mammalian cells.
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The <a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Characterisation">characterization</a> of the fungal promoters was successfully performed with an X-gal analysis, where the expression of ß-galactosidase results in blue colonies. Furthermore, quantitative ß-galactosidase and Bradford assays were performed in order to measure protein production and ß-galactosidase activity. Both promoters proved to be of medium strength.
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For demonstration of the Plug' Play with DNA system in <a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/mammalian">mammalian cell</a> lines, U-2 OS was transfected with plasmids containing genes encoding fluorescent proteins. The transfected cells capability to express the fluorescent proteins was investigated with confocal microscopy, which resulted in amazing <a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/mammalian">pictures</a> of fluorescent peroxisomes and coloured U-2 SO cells.
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Some of the submitted BioBricks have been successfully sequenced, and the data has been examined in order to verify that no mutations or incorrect insertions had occurred. The analysis of the sequencing data showed that all the sequenced Plug 'n' Play BioBricks had been correctly assembled without mutations. The sequencing and the characterization both showed that the system works as expected in <i>A. nidulans</i> and in the mammalian cell line U-2 OS.
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<a name="Fulfilment of medal requirements"></a><h1><b>Fulfilment of medal requirements</b></h1>
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<b>Bronze</b><br><br>
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<input type="checkbox" checked disabled="true"><b>Team registration</b><br><br>
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<input type="checkbox" checked disabled="true"><b>Complete Judging form</b><br><br>
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<input type="checkbox" checked disabled="true"><b>Team Wiki</b><br>
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We have made a description of our project (see The project), and submitted parts can be found under Results.<br><br>
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<input type="checkbox" checked disabled="true"><b> Present a poster and a talk at the iGEM Jamboree</b><br>
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We plan to make an excellent presentation and a magnificent poster that will be transported on first class from Denmark to Amsterdam right before the Jamboree. <br><br>
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<input type="checkbox" checked disabled="true"> <b>At least one new submitted and well-characterized standard BioBrick Part or Device. A new application of and outstanding documentation (quantitative data showing the Part’s/ Device’s function) of a previously existing BioBrick part in the “Experience” section of that BioBrick’s Registry entry also counts.</b><br>
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We have submitted 49 new <a hret="https://2011.igem.org/Team:DTU-Denmark-2/results/submitted_biobricks">BioBricks</a>.
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<b>Silver</b><br> <br>
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<input type="checkbox" checked disabled="true"><b>Demonstrate that at least one new BioBrick Part or Device of your own design and construction works as expected</b><br>
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We have demonstrated the function of the device, <a hret=!http://partsregistry.org/Part:BBa_K678002"> BBa_K678002</a>, that has been transfected into the mammalian cell line, U-2 OS. Furthermore, characterization of fungal promoter parts have been executed. These results can be viewed at our homepage or in the BioBrick registry for <a href="http://partsregistry.org/Part:BBa_K678000">BBa_K678000</a> and <a href="http://partsregistry.org/Part:BBa_K678001">BBa_K678001</a><br><br>
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<input type="checkbox" checked disabled="true"><b>Characterize the operation of at least one new BioBrick Part or Device and enter this information in the “Main Page” section of that Part’s/Device’s Registry entry.</b><br>
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<b>Gold</b> <br>
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<input type="checkbox" checked disabled="true"><b>Improve the function of an existing BioBrick Part or Device (created by another team or your own institution in a previous year) and enter this information in the Registry (in the “Experience” section of that BioBrick’s Registry entry), and don't forget to create a new registry page for the improved part.</b><br><br>
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<input type="checkbox" checked disabled="true"><b>Help another iGEM team by, for example, characterizing a part, debugging a construct, or modeling or simulating their system.</b><br>We have helped the iGEM team Copenhagen with the design and assembly of some of their <a href="https://2011.igem.org/Team:Copenhagen/Project/Data">BioBricks</a>.<br><br>
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<input type="checkbox" disabled="true"><b>Outline and detail a new approach to an issue of Human Practice in synthetic biology as it relates to your project, such as safety, security, ethics, or ownership, sharing, and innovation.</b><br><br>
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Even though we first started the iGEM project in the end of June, we have managed to design a fully standardized cloning system called Plug'n Play with DNA. We think that iGEM should be about fast and easy assembling of BioBricks, which led us to use features from the USER fusion standard assembly to model our <a hret="https://2011.igem.org/Team:DTU-Denmark-2/Project/overview"> Plug'Play with DNA </a>assembly system. Our system consist of:
 
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<li> Ready PCR products - All ready to use!
 
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<li> Back-up plasmide - To ensure mutation free amplification.
 
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<li> Guide on customization - All procedures only require 1 round of PCR and assembly
 
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To proof our concept, we have characterized the two fungal promoters, PalcA and DMKP-P6,  and ___ mammalian promoters, ________ __________ . The characterization of the fungal promoters was performed by evaluation of the qualitative X-gal analysis, where the expression of ß-galactosidase results in blue colonies. Furthermore, quantitative ß-galactosidase and Bradford assay was performed to measure the protein production.
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Latest revision as of 00:38, 22 September 2011




Achievements

Achievements


The team came together during a course in mammalian cell biology in June, and that was when the idea of iGEM popped into our heads. Even though we started the iGEM project late June, we have managed to design a fully standardized cloning system called Plug'n Play with DNA. We thought that time should be spent on making cool iGEM projects, instead of struggling with unwanted restriction sites and cumbersome assembly systems. This led us to standardize USER fusion by designing the Plug'n'Play with DNA assembly system.


Our Plug 'n' Play with DNA kit consist of:
  • 49 ready-to-use BioBricks.
  • 21 backup plasmids.
  • A quick 'n' easy guide on how to customize the standard

    • To ensure that the BioBricks and devices have been assembled correctly we have tested the functionality of them in Aspergillus nidulans and in mammalian cells. We have characterized the two fungal promoters, PalcA and DMKP-P6, and demonstrated function of the cytomegalovirus (CMV) promoter in mammalian cells. The characterization of the fungal promoters was successfully performed with an X-gal analysis, where the expression of ß-galactosidase results in blue colonies. Furthermore, quantitative ß-galactosidase and Bradford assays were performed in order to measure protein production and ß-galactosidase activity. Both promoters proved to be of medium strength. For demonstration of the Plug' Play with DNA system in mammalian cell lines, U-2 OS was transfected with plasmids containing genes encoding fluorescent proteins. The transfected cells capability to express the fluorescent proteins was investigated with confocal microscopy, which resulted in amazing pictures of fluorescent peroxisomes and coloured U-2 SO cells. Some of the submitted BioBricks have been successfully sequenced, and the data has been examined in order to verify that no mutations or incorrect insertions had occurred. The analysis of the sequencing data showed that all the sequenced Plug 'n' Play BioBricks had been correctly assembled without mutations. The sequencing and the characterization both showed that the system works as expected in A. nidulans and in the mammalian cell line U-2 OS.



    Fulfilment of medal requirements

    Bronze

     Team registration

    Complete Judging form

    Team Wiki
    We have made a description of our project (see The project), and submitted parts can be found under Results.

    Present a poster and a talk at the iGEM Jamboree
    We plan to make an excellent presentation and a magnificent poster that will be transported on first class from Denmark to Amsterdam right before the Jamboree.

    At least one new submitted and well-characterized standard BioBrick Part or Device. A new application of and outstanding documentation (quantitative data showing the Part’s/ Device’s function) of a previously existing BioBrick part in the “Experience” section of that BioBrick’s Registry entry also counts.
    We have submitted 49 new BioBricks.

    Silver

     Demonstrate that at least one new BioBrick Part or Device of your own design and construction works as expected
    We have demonstrated the function of the device, BBa_K678002, that has been transfected into the mammalian cell line, U-2 OS. Furthermore, characterization of fungal promoter parts have been executed. These results can be viewed at our homepage or in the BioBrick registry for BBa_K678000 and BBa_K678001

    Characterize the operation of at least one new BioBrick Part or Device and enter this information in the “Main Page” section of that Part’s/Device’s Registry entry.


    Gold

    Improve the function of an existing BioBrick Part or Device (created by another team or your own institution in a previous year) and enter this information in the Registry (in the “Experience” section of that BioBrick’s Registry entry), and don't forget to create a new registry page for the improved part.

    Help another iGEM team by, for example, characterizing a part, debugging a construct, or modeling or simulating their system.
    We have helped the iGEM team Copenhagen with the design and assembly of some of their BioBricks.

    Outline and detail a new approach to an issue of Human Practice in synthetic biology as it relates to your project, such as safety, security, ethics, or ownership, sharing, and innovation.

    Retrieved from "http://2011.igem.org/Team:DTU-Denmark-2/Project/achievements"