Part:BBa_K516214:Experience

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••••• UNIPV-Pavia iGEM 2011

Characterized with: BBa_K516210 pTet-RBS30-LuxI BBa_K516211 pTet-RBS31-LuxI BBa_K516212 pTet-RBS32-LuxI Though these parts don't have a transcriptional terminator, they have been characterized in low copy plasmid pSB4C5, that contains the BBa_B0054 terminator. This choice is motivated by the need to reproduce the exact experimental context of the final circuit, as described in solution section. LuxI has been characterized in terms of enzymatic activity under the regulation of pTet promoter. KM,LuxI and Vmax parameters representing its activity have been estimated and the promoter strength (represented by a synthetic parameter αpTet for every pTet-RBS combination) at full induction (100 ng/ml) has been estimated too with a simultaneous fitting of the available data. LuxI has been characterized through the Biosensor BBa_T9002 (see modelling section). The HSL synthesis rate has been evaluated according to the model equations, properly adjusted. The parameters Vmax, kM,LuxI and αRBSx were estimated with a simultaneous fitting of the data collected as described in measurement section for the four measurement parts pTet-RBSx-LuxI-TT assayed by BBa_T9002 biosensor section. The estimated parameters for the enzymatic activity of LuxI are reported in the table below: Vmax kM,LuxI αB0030 αB0031 αB0032 αB0034 3.56*10-9 6.87*103 87 8.5 ND 252 The collected data have been used to identify the parameters of our model. Despite the data-poor context, the model predictions fit the experimental data, thus demonstrating that the equation that models the HSL synthesis by LuxI is a good approximation of real processes.

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