Team:KULeuven/Data

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Data page

How does E.D. Frosti work?

The first illustration is a global overview of our project in which you can see how the plasmids (and the parts) function in our system. The next illustration shows the plasmids more in detail, with the favorite new parts and the characterized pre-existing parts.


Plasmid 1 contains BBa_K584028 that enables E.D. Frosti to immediately respond to lactose and produces INP. BBa_K584027 is a precursor of BBa_K584028 with a constitutive promoter.
Plasmid 2 can contain BBa_K584026 to test the activity of AFP. Due to lack of time we could not construct a biobrick in which the pBAD promoter is linked to the AFP.
Plasmid 3 contains BBa_K584004. This is is a GFP-construct to test the activity of the HybB promoter. Due to lack of time we could not test this biobrick. We wanted to engineer a biobrick in which the ribokey is controlled by the HybB promoter. BBa_K584027 was inserted in plasmid 1 to characterize INP.

Data for our favorite new parts

1. BBa_K584028 (Main Page) - pLac-Lux hybrid promoter + INP , BBa_K584028 (K.U.Leuven, iGEM 2011): Tested with MilliQ water (first purified via reversed osmosis over 0,45 µm then filtered through a 0,22µm filter to remove all nucleating agents) at -6°C. We added 1mM IPTG to E.D. Frosti's medium. Water freezes immediately upon addition. Addition of a control (GFP construct) E.D. Frosti did not result in freezing. Note: The promoter coupled to GFP did not show any leakiness, but when we coupled the promoter to the INP we noticed a certain leakiness. The dual inhibition system is a way to get around this leakiness.

2. BBa_K584027 (Main Page) - Constitutive Promoter + INP , BBa_K584027 (K.U.Leuven, iGEM 2011): Tested with MilliQ water (first purified via reversed osmosis over 0,45 µm then filtered through a 0,22µm filter to remove all nucleating agents) at -6°C. Water freezes immediately upon addition. Addition of a control (GFP construct) E.D. Frosti did not result in freezing.

3. BBa_K584026 (Main Page) - constitutive promoter + AFP , BBa_K584026 (K.U.Leuven, iGEM 2011): We used the same conditions for the MilliQ water, but now at -10°C. Unfortunately addition of E.D. Frosti coated with AFP (or nothing: GFP control) did not result in prevention of ice formation. Due to lack of time we could not test other conditions (different temperatures, different concentrations,...)

Data for pre-existing parts

1. BBa_I13453 (Experience) - pBAD promoter, BBa_I13453 (Endy Lab, iGEM 2005): Fused a GFP reporter to this promoter (which created biobrick: BBa_K584000). The results can be found at the experience page of that promoter.

2. BBa_J23119 (Experience) - constitutive promoter, BBa_J23119 (Berkeley, iGEM 2006): Fused a GFP reporter to this promoter (which created biobrick: BBa_K584001). The results can be found at the experience page of that promoter.

We've also characterized the following parts

1. BBa_K584004 (Main Page) - HybB promoter + GFP generator, BBa_K584004 (KULeuven, iGEM 2011): We created this biobrick but due to lack of time we could not test it.