Team:BIT/BA2
Necessity of E.coli ΔLysA
After we have verified that lysine can be the signal upstream the sensor, what we need to do next is to conform the connection between the lysine concentration and the growth of E.coli ΔLysA. Before that, we have to make sure that the growth of wild type E.coli has no connection with lysine concentration. Part BBa_K876070 was transformed into wild type E.coli to test its OD600 and GFP fluorescence every hour with microplate reader. Lysine was added in increments of 0.5ug/mL to establish a range of 0-3ug/mL. It shows that the growth of wild type E.coli has no connection with lysine concentration. From this, we can say that it’s necessary for E.coli ΔLysA to be the signal receiver of lysine.
Confirmation in Models
After we have confirmed that the E.coli ΔLysA can response and transform lysine signal to the fluorescent signal, what we need to do is to amplify the fluorescent signal and improve the sensitivity. To solve this problem and make sure that our own-designed circuit works well, we need to test the effect of the strong promoter, cyclic amplifier and dual fluorescence system respectively. The verified experiments are as follows.
Strong Promoter
To confirm that the strong constitutive promoter can induce plux promoter, we designed the biobrick BBa_K2305004. The purpose of this biobrick is to produce as much green fluorescence as possible by inducing the induced promoter plux, to ensure a stronger expression in E.coli.
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