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

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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.   
 
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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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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The parameters N<sub>max</sub>, &mu; and &gamma;<sub>HSL</sub> are known and summarized in the table below:</p>
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Latest revision as of 17:00, 24 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

UNIQfc9a98a731475e09-partinfo-00000000-QINU UNIQfc9a98a731475e09-partinfo-00000001-QINU


UNIQfc9a98a731475e09-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:



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 BBa_T9002 experience page).
The HSL synthesis rate has been evaluated according to the model equations, properly adjusted. In particular, the ODE system describing the behavior of this measurement circuit is reported here:



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 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 through BBa_T9002 biosensor - experience page.

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
ND=Not Detected

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.

The provided parameters kM and Vmax represent the enzymatic activity of LuxI, described by our model. They must not be confused with the operative parameters of the Michaelis-Menten relation. These synthetic parameters have a great importance, since they can be used in more complicated models in order to predict the behavior of complex circuits.

UNIQfc9a98a731475e09-partinfo-00000005-QINU