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Team:Groningen/project memory units
From 2011.igem.org
Memory Unit
As a memory units we are using bistable autoinducing loops that can be suppressed via reversed promoter jammer. Autoinducing loop play a role as an amplifier of input signal by regulating its own expression. To switch it off, we adopted the endogenous device from British Columbia 2009 team, which uses a reverse antisense promoter downstream a coding sequence (https://2009.igem.org/Team:British_Columbia/Jammer).
Our implementation
In our implementation the first memory unit is based around a modified lambda Prm promoter(Bba_I12006) producing its own inducer cI. By means of the jammer is can be repressed with LasT. The second memory unit uses the lasB promoter(BBa_R0079) producing its own inducer LasR. This second loop can be repressed by arabinose. Both our cI gene and our LasR have degredation tag that help us tweak their bistable behavoir. To study the dynamics of the expression and half-life of different autoinducing loop components, the transcription factor genes (cI and LasR) where fused in combinations with various RBS's and degradation tags, resulting 12 different construct variants.
Results
TODO Firstly, we would like to test the leakage of promoter by cotransform autoinducing loop plasmid and reporter plasmid under control of appropriate promoter. Secondly, cotransformation assembled input and output systems with plasmid bearing various-variants of autoinducing loops would provide information about memory unit behaviour in time. Finally, integration of experimental data and our model would lead us to fine-tuned the expected system behaviour.
