Part:BBa_C0060:Experience

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UNIQ5ee432bc24344243-partinfo-00000000-QINU

••••• UNIPV-Pavia iGEM 2011	NB: unless differently specified, all tests were performed in E. coli MGZ1 in M9 supplemented medium at 37°C in low copy plasmid pSB4C5. The AiiA enzyme activity has been characterized under the regulation of ptet promoter, assaying its enzymatic activity, with these parts: BBa_K516220 pTet-RBS30-AiiA-TT BBa_K516221 pTet-RBS31-AiiA-TT BBa_K516222 pTet-RBS32-AiiA-TT BBa_K516224 pTet-RBS34-AiiA-TT A system of differential equations has been derived and we tried to identify the parameters, studying the exponential growth phase: The four measurment parts ptet-RBSx-LuxI were quantitatively characetrized using the BBa_T9002 biosensor. Unfortunately we were not able to estimate the model parameters (kcat, kM,AiiA) because placing the device in low copy plasmid (pSB4C5) we did not observe HSL degradation, as you can see in the figures below: We were only able to give semi-quantitative information about AiiA enzyme activity, performing an experiment in E. coli TOP10, with the four measurement devices in pSB1A2; the results are summerized in the graphs below: In this way we can give a measure of AiiA activity as the percentage of HSL degraded in 7 hours (measurements provided as average [stdandard deviation]): Measurement device HSL degraded percentage BBa_K516220 96.11 [3.66] BBa_K516221 77.60 [2.66] BBa_K516222 50.66 [16.87] BBa_K516224 100 [0] In order to evaluate the RBSs efficiency, we compute the ratio between the percentage of the degraded HSL realtive to the measurement device with the standard RBS (BBa_B0034):
RBS	RBS efficiency
BBa_B0030	0.96
BBa_B0031	0.78
BBa_B0032	0.51
BBa_B0034	1