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Team:Groningen/project memory units
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| - | + | === Memory Unit === | |
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| + | Our system composed of 2 memory units, consisting autoinducing loop and jammer-based off switch. Autoinducing loop play a role as an amplifier of input signal by regulating its own expression. To swith 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). First memory unit contain cI gene downstream modified lambda PRM promoter (Bba_I12006) and flanked-terminators in forward and reverse direction. This system is induced by CI (lambda repressor) and repressed by LasR. Whereas, second memory unit contain lasR gene downstream LasB promoter (BBa_R0079), which induced by LasR and repressed by arabinose. To study the dynamic expression and half-life of autoinducing loop components, which best fitted the expected system, the transcription factor gene (cI and LasR) was fused in combination with various RBS and degradation tag, resulting 12 different constract variants. 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-variant of autoinducing loops would lead us to obtain 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. | ||
Revision as of 19:42, 3 September 2011
Memory Unit
Our system composed of 2 memory units, consisting autoinducing loop and jammer-based off switch. Autoinducing loop play a role as an amplifier of input signal by regulating its own expression. To swith 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). First memory unit contain cI gene downstream modified lambda PRM promoter (Bba_I12006) and flanked-terminators in forward and reverse direction. This system is induced by CI (lambda repressor) and repressed by LasR. Whereas, second memory unit contain lasR gene downstream LasB promoter (BBa_R0079), which induced by LasR and repressed by arabinose. To study the dynamic expression and half-life of autoinducing loop components, which best fitted the expected system, the transcription factor gene (cI and LasR) was fused in combination with various RBS and degradation tag, resulting 12 different constract variants. 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-variant of autoinducing loops would lead us to obtain 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.
