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Team:DTU-Denmark-2/Project/achievements
From 2011.igem.org
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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 | + | 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 Plug'n Play assembly consist of: |
<li> 49 BioBricks and 21 plasmids- All ready to use!. | <li> 49 BioBricks and 21 plasmids- All ready to use!. | ||
<li> Back-up plasmide - To ensure mutation free amplification. | <li> Back-up plasmide - To ensure mutation free amplification. | ||
| - | <li> Guide on customization - All procedures only require 1 round of PCR and assembly | + | <li> Guide on customization - All procedures only require 1 round of PCR and 1 round of assembly. |
<br><br> | <br><br> | ||
| - | To verify the function of our design we examined various Biobriks, parts and devices, by experimental procedures described under protocols. | + | To verify the function of our design we examined various Biobriks, parts and devices, by experimental procedures described under <a href=" https://2011.igem.org/Team:DTU-Denmark-2/Team/Protocols" >protocols</a>. <br> |
We have characterized the two fungal promoters, PalcA and DMKP-P6, and proved function of three mammalian promoters, SV40, PGK, CMV. <br> | We have characterized the two fungal promoters, PalcA and DMKP-P6, and proved function of three mammalian promoters, SV40, PGK, CMV. <br> | ||
The <a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Characterisation">characterization</a> of the fungal promoters was qualitative evaluated with an X-gal analysis, where the expression of ß-galactosidase resulted in blue colonies for both PalcA and DMKP-6. Furthermore, quantitative ß-galactosidase and Bradford assay was performed to measure the protein production and ß-galactosidase activity. The activity of ß-galactosidase was highest for DMKP-6 than PalcA, which was expected to have same activity.<br> | The <a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Characterisation">characterization</a> of the fungal promoters was qualitative evaluated with an X-gal analysis, where the expression of ß-galactosidase resulted in blue colonies for both PalcA and DMKP-6. Furthermore, quantitative ß-galactosidase and Bradford assay was performed to measure the protein production and ß-galactosidase activity. The activity of ß-galactosidase was highest for DMKP-6 than PalcA, which was expected to have same activity.<br> | ||
Revision as of 17:20, 17 September 2011
Achievements
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 Plug'n Play assembly consist of:
To verify the function of our design we examined various Biobriks, parts and devices, by experimental procedures described under protocols.
We have characterized the two fungal promoters, PalcA and DMKP-P6, and proved function of 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 resulted in blue colonies for both PalcA and DMKP-6. Furthermore, quantitative ß-galactosidase and Bradford assay was performed to measure the protein production and ß-galactosidase activity. The activity of ß-galactosidase was highest for DMKP-6 than PalcA, which was expected to have same 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
Through this study, we have registered 70 parts.... Name part -favorite.
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 BBa_K678001.
Gold
1:
2: We have collaborated with iGEM Copenhagen, where we help designing and constructing some of their BioBricks with our Plug'nPlay system.
3:
