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Team:Groningen/project characterisation mu
From 2011.igem.org
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 | |||||||||||||||||
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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). |
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| 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 | |||||||||||||||||
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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). |
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| Only one population is visible blabla | |||||||||||||||||
