Achievements

The team came together during a course in mammalian cell biology in June, and that's when the idea of iGEM popped into our heads. Eventhough we started the iGEM project in 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 systems, instead of struggeling with unwanted restriction sites and limited assembly systems. This led us to the USER fusion standard assembly as a model our Plug'Play with DNA assembly system.

49 ready-to-use BioBricks.

21 backup plasmids.

A quick 'n' easy guide on how to customize new BioBricks or introducing specific mutations

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 proved the function of the cytomegalovirus (CMV) promoter for 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 for fluorescence molecules. The transfected cells capability to express the fluorescence molecules was investigated with confocal microscopy, which resulted in amazing pictures of glowing peroxisomes and colored U-2 SO cells. Some of the submitted BioBricks have been successfully sequenced, and the data have been examinated in order to verify that no mutations or incorrect insertions had occurred. The analysis of the sequencing data showed that the Plug 'n' Play parts had been correctly assembled. The sequencing and the characterization both show that the system works as expected in A. nidulans and in the mammalian cell line U-2 OS without unwanted insertion and mutations.

Fullfillment of medal requirements

Bronze
1. Team Registration
All team members are registered

2. Complete judging form
Judgingform will be completed as soon as possible.

3. Create and share a description of the teams project using iGEM wiki and the Team's parts.
We have made a description of our project (see The project), and submitted parts can be found under Results.

4. Plan to present a poster and 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.

5. Enter information detailing at least one new standard BioBrick Part or Device in the Registry of Standard Biological Parts
We have submitted 49 new BioBricks.


Silver
1+2. Demonstrate that at least one new BioBrick Part or Device of your own design and construction works as expected; characterize the operation of your new part/device. Enter this information on the "Main Page" section of the registry
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


Gold
1:Improve an existing BioBrick Part or Device and enter this information back on the Experience Page of the Registry.
We used the CMV promoter, BBa_J52034 , as the regulator for our fluorescence reporters, and it worked well. The promotor and our experience is described in Experience.

2: 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.