Difference between revisions of "Team:ColegioFDR Peru/Project"

Construct Overview

The first accumulator construct is a theoretical biobrick in which we attempted to make using BBa_K1355003 designed by the iGEM14_UFAM_BRAZIL team. Our goal was to introduce a Green Fluorescence Protein part before the terminator of the mercury accumulating device. The first problem that we encountered was that there were no restriction sites present inside the biobrick so we could not open the plasmid. In order to insert the GFP, we used the overlapping PCR protocol to insert the GFP. The way that it would have worked was by designing a primer that had an extra sequence attached that would hang freely when the primer is attached. This free hanging sequence would code for the GFP. We would use that GFP primer as a primer for the accumulator device so that when the DNA polymerase reads and copies the device sequence, it would add on the extra GFP sequence. Unfortunately, after three tried of this method, we were unable to get good results.

The Killer Red Construct is a double condition suicide sequence. The first condition is the presence of mercury. When the bacteria are in an environment containing mercury, the production of RBS (BBa_B0030) is initiated through the Promotor (BBa_K346002). Then, the Killer Red protein (BBa_K1184000) is produced. The second condition is the presence of light. In order for the Killer red to activate, light (540-585 nm) needs to be shined onto the bacteria to initiate the reaction.