Team:DTU-Denmark-2/Project/achievements

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Achievements

  • We achieved to design a standardized cloning system, called Plug'n Play with DNA, for both mammalian cells and fungi.
  • We characterized BioBricks with ß-galactosidase assays, Bradford assay and fluorescence
  • We submitted 73 parts to the Registry of Standard Biological Parts requirements listed. 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 Plug'Play with DNA assembly system. Our ready system consist of:
  • 49 BioBricks and 21 plasmids- All ready to use!.
  • Back-up plasmide - To ensure mutation free amplification.
  • Guide on customization - All procedures only require 1 round of PCR and assembly

    To verify the function of our design we examined various Biobriks, parts and devices.
    We have characterized the two fungal promoters, PalcA and DMKP-P6, and three mammalian promoters, SV40, PGK, CMV.
    The characterization of the fungal promoters was qualitative evaluated with an 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 and ß-galactosidase activity.
    To demonstrate Plug' Play with DNA also was applicable in mammalian cell lines, the transfected cells capability to express the fluorescence molecules was investigated with confocal microscopy.


    To ensure that there had been no occurrence of mutation and incorrect insertions in the BioBricks, they was sequenced and the resulting result was examined. The results from the sequencing supported the evaluation and characterization, implying that the system works as expected in both mammalian cells and fungi

    Medal accomplishment
    Bronze
    1-3: Team registration, judging form and team wiki was completed in time. 4: Presentation is taken form and the rehearsal for the talk at iGEM is getting fine-tuned. 5: We used the CMV promoter, BBa_J52034 , as the regulator for our fluorescence reporters, and it worked well. Announced under Experience Silver 1-2: We have, among other, demonstrated the function of the device, BBa_K678002, transfected into mammalian cell lines. Furthermore, characterization of fungal promoter parts has been executed, and the results can be viewed at our homepage or in the BioBrick registry for BBa_K678000 and Retrieved from "http://2011.igem.org/Team:DTU-Denmark-2/Project/achievements"