Team:KULeuven/Biobricks

Biobricks

The biobricks that we used are on the iGEM registry which can be found here. Underneath you can see several short videos in which we added different kinds of E.D Frosti to supercooled (-6°C) ultrapure “milli-Q” water (Millipore Corporation). The first video (GFP-control), is a control: E.D. Frosti only contains a GFP construct. When we add it to the supercooled water, it does not freeze.

In the second video (IPTG), E.D. Frosti contains BBa_K584028 which contains the INP gene, coupled to the lactose inducible promoter. We added 1mM IPTG to E.D. Frosti. When we added it to supercooled water, it froze immediately, prooving the functionallity of our biobrick.

In the third video (INP), E.D. Frosti contains a constitutively active INP producing biobrick ( BBa_K584027). When we added this to the supercooled water, it froze immediately.



The fourth video (No IPTG) shows what happened when we added E.D. Frosti with the BBa_K584028 biobrick, but that did not receive a lactose (IPTG) stimulus. Although our GFP-characterization of this biobrick showed that the promoter was not leaky, the water still froze. This can have several causes, two examples are: the coupling of the INP gene to the promoter made the system leaky. It is also possible that there is a very low (basal) expression of the INP gene and that this small amount of INP is enough to induce ice nucleation.