After we confirmed the construction of our device we start planning our characterization tests to prove that our device really works. To fulfill this aim, our new device was transformed in E. coli C43 strain, an overexpression strain in which the T7 promoter can be induced by IPTG. We transformed cells with our device, BBa_K284022, and with the BBa_K112806(without promoter).
A transformed colony was incubated in LB medium with ampicillin at 37ºC overnight. The culture was diluted in new LB medium with ampicillin to an OD=0,2 and incubated at 37ºC until reach an OD=0,8. At this point, the cells were induced with IPTG 1 mM and incubated again for 4 hours. After the end of the induction period the cells were plated in LB agar with ampicillin in order to confirm the diminished cellular growth. To have a definitive confirmation, we also performed SDS-PAGE to notice the protein overexpression. The results are shown in ColiGuard results.
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