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 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 that induce the 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, lactose or arabinose will generate a ribolock-ceaB construct ( BBa_K584015 and BBa_K584016).The Ribokey can be activated by a cold shock( 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_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 ) and the lactose-inducible promoter (yielding BBa_K584028).

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

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.

3. BBa_K091100 - Lac-Lux hybrid promoter (Davidson-Missouri_Western, iGEM 2008): fused a GFP to this promoter (which created biobrick BBa_K584002 - constitutive promoter). 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_K091107 - pLux/cI Hybrid Promoter (K.U.Leuven, iGEM 2011): We wanted to use this promoter at the inception of our project, however on a 1% agarose gel we saw one fragment of a different height than reported by previous iGEM competitions.

3. BBa_K091104 - pLac/Mnt Hybrid Promoter (K.U.Leuven, iGEM 2011): We wanted to use this promoter at the inception of our project, however on a 1% agarose gel we saw one fragment at a different height than reported by previous iGEM competitions.