Team:KULeuven/Data

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 in more detail, with the favorite new parts and the characterized pre-existing parts.


Plasmid 1 contains the bricks necessary to induce expression of the red color (via the CrtEBI system) and ice-nucleating protein (INP) in a lactose-dependent manner. For this plasmid, the INP has been cloned (BBa_K584024) and tested for functionality by putting it behind the constitutive (BBa_K584027) and lactose-inducible ( BBa_K584028 ) promoter.

Plasmid 2 contains the bricks necessary to induce expression of the black color (via the melA system) and antifreeze protein (AFP) in a arabinose-dependent manner. For this plasmid, the AFP has been cloned ( BBa_K584020) and tested for functionality by putting it behind the constitutive promoter (BBa_K584026).

Plasmid 3 contains the necessary bricks for our cell death mechanism. Briefly, both lactose and arabinose will generate a ribolock-ceaB construct ( BBa_K584015 and BBa_K584016), which can be activated by a cold shock induced-expression of the ribokey ( BBa_K584014). For this plasmid we cloned the lactose- and arabinose-inducible ceaB construct ( BBa_K584016 and BBa_K584015 resp.) and characterized the cold shock promoter using the Biobrick BBa_K584004. BBa_K584027 was inserted in plasmid 1 to characterize INP.

Data for our favorite new parts

1. BBa_K584020 - AFP extracellular protein generator (K.U.Leuven, iGEM 2011): this biobrick generates AFP that will be expressed extracellularly. We tested this biobrick by putting it behind a constitutive promoter (which yielded BBa_K584026 ).

2. BBa_K584024 - INP generator (K.U.Leuven, iGEM 2011): this biobrick generates INP that is expressed extracellularly. We tested this biobrick by putting it behind a constitutive promoter (which yielded BBa_K584027).

Data for pre-existing parts

1. BBa_I13453 - pBAD promoter (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 the promoter.

2. BBa_J23119 - constitutive promoter (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 the promoter.

We've also characterized the following parts

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

2. BBa_K584027, Constitutive promoter + INP (K.U.Leuven, iGEM 2011): we characterized this promoter by coupling it in front of a GFP. The characterization can be found on the registry page of the original biobrick: BBa_J23119 .

3. BBa_K584026, Constitutive promoter + AFP (K.U.Leuven, iGEM 2011): For this biobrick, we used the same constitutive promoter ( BBa_J23119 ), the characterization of this biobrick was the same.