Team:Bordeaux/Results

iGEM - Bordeaux - Biology - Results

We successfully obtain the biobricks needed for our project by transformation. As shown in figure one, our project requires 20 parts to assemble to reach our goal. As a lot of cloning is required, we made a cloning plan (fig.1). There were three steps to assemble properly each operon and one more step to pair the operons in order to have only two plasmids to insert during a final transformation.

Figure 1: from biobricks to operon. The first line shows the parts we get from the iGEM. The small squares represent an assembly step.

This plan shows that 20 assemblies must be done to achieve the work. As we progress through time, we experienced some troubles. At first, the transformation efficiency did not allow to obtain a lot of new parts. After troubleshooting, we proceed as described in the material and methods section. Nevertheless, time runs out fast, and figure 2 displays our work process at the time.

Figure 2: assembly processing. Boxes in green are for part or assembly we get. Red boxes symbolize the missing one either because the assembly won’t work, either because the previous parts are not available yet.

This allows us to present 5 new biobricks this year (our biobricks), with the hope it will help other teams to complete their project in the future year.
Considering the short amount of time left, we decided to aim at something a little smaller. As the operon I was the hardest to get, we gave up on it and decide to continue with only three operons and thus, fewer parts to assemble (fig 3).

Figure 3: A: operon 1 is suppressed, and operon 2 promoter is replaced with part of the promoter from operon 1 (shown in blue to underline the change). B: new regulation pathway, justifying some biobricks elimination. Bold pink arrows represent the signals sent to the neighbor cells, black lines represent internal signal sent.

Our main goal, ie creating a regulation network in a unicellular organism, remains the priority. Only the no more necessary biobricks were eliminated from the pathway. So, instead of having three colors on the dishes, only two should appear. Moreover, our first project provided some assemblies we were able to reuse in our attempt to make our project work. At the time we submit these results, the constructions are not over yet, as shown in figure 4.

Figure 4: assembly processing of the reduce project. Boxes in green are for part or assembly we get. Red boxes symbolize the missing one either because the assembly won’t work, either because the previous parts are not available yet.

iGEM - Bordeaux - Biology - Conclusion

2012 is the first year a team enter the iGEM competition in Bordeaux. All volunteers were eager to have fun and learn from the experience. At least, this part of the whole project was fulfilled.
The iGEM competition requires a lot of lab work, but also a lot of research before getting started. Even if we didn’t complete everything we aimed at, we hope the submitted biobricks will help other teams to realize their projects in the future years. We provided to the iGEM registry functional transcription units we obtained, and an operon under the control of two different promoters. As we feel we are very close, we will carry on with our experiments in the wish to really finish this project before starting another one for the 2013 edition !