Difference between revisions of "Part:BBa K516210:Experience"

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LuxI has been characterized in terms of enzymatic activity under the regulation of p<sub>Tet</sub> promoter. <br>
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K<sub>M,LuxI</sub> and V<sub>max</sub> parameters representing its activity have been estimated and the promoter strength (represented by a synthetic parameter &alpha;<sub>pTet</sub> for every pTet-RBS combination) at full induction (100 ng/ml) has been estimated too with a simultaneous fitting of the available data. <br>
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Since the measurement systems are only assayed in the exponential growth phase, in the equation (3) N &lt; N<sub>max</sub> is assumed.
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LuxI has been characterized through the Biosensor <partinfo>BBa_T9002</partinfo>. 
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The parameters N<sub>max</sub>, &mu; and &gamma;<sub>HSL</sub> (see the <a href='http://2011.igem.org/Team:UNIPV-Pavia/Project/Results'>Results section</a> for more details) are known and summarized in the table below:</p>
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<br>The HSL synthesis rate has been evaluated according to the <a href='http://2011.igem.org/Team:UNIPV-Pavia/Project/Modelling#Equations_for_gene_networks'>model equations</a>, properly adjusted. <br>
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The parameters V<sub>max</sub>, k<sub>M,LuxI</sub> and &alpha;<sub>RBSx</sub> were estimated with a simultaneous fitting of the data collected as described in <a href='http://2011.igem.org/Team:UNIPV-Pavia/Measurements#LuxI'>measurement section</a> for the four measurement parts <a href='http://2011.igem.org/Team:UNIPV-Pavia/Parts/Characterized#pTetLuxI'>pTet-RBSx-LuxI-TT</a> assayed by <a href='http://2011.igem.org/Team:UNIPV-Pavia/Measurements#T9002'>BBa_T9002 biosensor</a> section.  <br>
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<div align="center">
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<table  align="center" class='data' title='parameter value'>
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<td class="row"><b>N<sub>max</sub></sub></b></td>
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<td class="row"><b>&mu;</b></td>
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<td class="row"><b>&gamma;<sub>HSL</sub></b></td>
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<td class='row' >1*10<sup>9</sup></td>
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<td class='row' >0.004925</td>
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<td class='row' >0</td>
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</table>
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</div>
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<p align='justify'>
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The parameters V<sub>max</sub>, k<sub>M,LuxI</sub> and &alpha;<sub>RBSx</sub> were estimated with a simultaneous fitting of the data collected as described in <a href='http://2011.igem.org/Team:UNIPV-Pavia/Measurements#LuxI'>measurement section</a> for the four measurement parts <a href='http://2011.igem.org/Team:UNIPV-Pavia/Parts/Characterized#pTetLuxI'>pTet-RBSx-LuxI-TT</a> assayed by <a href='http://2011.igem.org/Team:UNIPV-Pavia/Measurements#T9002'>BBa_T9002 biosensor</a> section.   
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<div align="justify"><div class="thumbinner" style="width: 850px;">
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<a href="https://static.igem.org/mediawiki/2011/4/40/Luxi_prod.jpg">
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<img alt="" src="https://static.igem.org/mediawiki/2011/4/40/Luxi_prod.jpg" class="thumbimage" width="85%" height="50%"></a></div></div>
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<div align="justify"><p>The estimated parameters for the enzymatic activity of LuxI are reported in the table below:</p></div>
 
<div align="justify"><p>The estimated parameters for the enzymatic activity of LuxI are reported in the table below:</p></div>
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<td class='row'><b>V<sub>max</sub></b></td>
 
<td class='row'><b>V<sub>max</sub></b></td>
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Revision as of 13:56, 23 September 2011

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K516210

User Reviews

UNIQb341c27baad5d4b7-partinfo-00000000-QINU UNIQb341c27baad5d4b7-partinfo-00000001-QINU


UNIQb341c27baad5d4b7-partinfo-00000002-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.

Characterized with:



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.

Since the measurement systems are only assayed in the exponential growth phase, in the equation (3) N < Nmax is assumed. The parameters Nmax, μ and γHSL (see the Results section for more details) are known and summarized in the table below:

Nmax μ γHSL
1*109 0.004925 0

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.

UNIQb341c27baad5d4b7-partinfo-00000005-QINU