Team:Gothenburg/Parts/Results

Results

In this section we present our laboratory victories!
Unfortunately, not all parts could be created and successfully transformed. All the genetic fragments on their own could be amplified and purified. Problems accured during the transformations into yeast. Several different transformation protocols were used but the success rate was disappointing.

Constructs

In the end only the two counting constructs could be isolated.
We were able to successfully transform the corresponding fragments into yeast cells, were the homologous recombinase activity merged them together. The positive results by growth on the selective medium were confirmed by a colony PCR.

Figure 1 and 2. UV-Picture of the colony PCR gel. The first seven bands in Figure 1 as well as the first four bands in Figure 2 correspond to the correct transformed colonies.

Finally, we also observed our transformed yeast cells under a fluorescent microscope to verify the function of the construct.

Figure 3. Fluorescent microscopy image of transformed yeast cells with the first construct. Daughter cells show a yellow fluorescence.

Figure 3 shows clearly yellow fluorescence, stating not only the successful transformation but also confirms the function of the ribozymes. Without the cleavage caused by them the fluorescence protein would not be folded properly and no fluorescence would be detectable.
As our construct shows fluorescence in the daughter cells but not in the mother cells we know that the cyclin activation is functioning and also that the degradation tag (D-tag) also is working: The D-tag is our submission to the parts registry.

One of the major issues we encountered was a problem in our concept plan itself: To make the desired process work all of the constructs need to be created and to be transformed into one cell. Successful transformations of a single part into one cell do not proof the concept.