Memory Units

cI Autoinducing Loops

The autoinducing loop driven by cI will function as our first memory unit. We aimed to find a combination of RBS, transcription factor and degradation tag that enables us to toggle between the two steady states in a robust way. To estimate the rate in which single cells are activated by too much leakage of cI protein we measured the fluorescence in uninduced cells with a flow cytometer. To measure the activation we used a construct with the PRM promoter producing GFP with an LVA tag to make dynamic measurement possible.

cI autoinducing loop 1 (BBa_K00) after 4 hours of incubation
Figure 1. A dotblot of uninduced cells carrying the pSB1K3 plasmid with the biobrick BBa_ and the plasmid pSB1A3 with as well an input(PBAD controlling cI) as well as the output (PRM controlling GFP-LVA).	Number  % of total Mean fluorescence Visible 50000 100 --- Fluorescent 9180 18,36 11419 Non-Fluorescent 34424 68 5974
Two populations are visible on the dotblot. We seperated both populations on the histogram

cI autoinducing loop 6 (BBa_K00) after 4 hours of incubation
Figure 2. A dotblot of uninduced cells carrying the pSB1K3 plasmid with the biobrick BBa_ and the plasmid pSB1A3 with as well an input(PBAD controlling cI) as well as the output (PRM controlling GFP-LVA).	Number  % of total Mean fluorescence Visible 50000 100 --- Fluorescent 9180 18,36 11419 Non-Fluorescent 34424 68 5974
Only one population is visible blabla