Difference between revisions of "Part:BBa C0060:Experience"

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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
how you used this part and how it worked out.
 
how you used this part and how it worked out.
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===Applications of BBa_C0060===
 
===Applications of BBa_C0060===
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A system of differential equations has been derived and we tried to identify the parameters, studying the exponential growth phase:
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A system of differential equations has been derived and parameters would have been identified, studying the exponential growth phase:
  
 
<div style='text-align:justify'>
 
<div style='text-align:justify'>
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The four measurment parts ptet-RBSx-LuxI were quantitatively characetrized using the <a href="https://parts.igem.org/Part:BBa_T9002:Experience#t9002">BBa_T9002 biosensor</a>. Unfortunately we were not able to estimate the model parameters (k<sub>cat</sub>, k<sub>M,AiiA</sub>) because placing the device in low copy plasmid (<a href="https://parts.igem.org/Part:pSB4C5">pSB4C5</a>) we did not observe HSL degradation, as you can see in the figures below:
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The four measurment parts ptet-RBSx-LuxI were quantitatively characetrized using the <a href="https://parts.igem.org/Part:BBa_T9002:Experience#t9002PV11">BBa_T9002 biosensor</a>. Unfortunately the model parameters (k<sub>cat</sub>, k<sub>M,AiiA</sub>) were not estimated because placing the device in low copy plasmid (<a href="https://parts.igem.org/Part:pSB4C5">pSB4C5</a>)resulted in no HSL degradation, as shown in the figures below:
  
 
<table width='100%'><tr><td width='50%'>
 
<table width='100%'><tr><td width='50%'>
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</table>
 
</table>
  
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We were only able to give semi-quantitative information about AiiA enzyme activity, performing an experiment in <em>E. coli</em> TOP10, with the four measurement devices in <a href="https://parts.igem.org/Part:pSB1A2">pSB1A2</a>; the results are summerized in the graphs below:
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A semi-quantitative information about AiiA enzyme activity is given, performing an experiment in <em>E. coli</em> TOP10, with the four measurement devices in <a href="https://parts.igem.org/Part:pSB1A2">pSB1A2</a>; the results are summerized in the graphs below:
  
 
<table width='100%'><tr><td width='50%'>
 
<table width='100%'><tr><td width='50%'>
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</table>
 
</table>
  
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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]):
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In this way a measure of AiiA activity is given, as the percentage of HSL degraded in 7 hours (measurements provided as average [stdandard deviation]):
  
 
<div align="center">
 
<div align="center">
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<table class="data">
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<table class="data" border="1">
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<tr><td class="row"><b>Measurement device</b></td><td><b>  HSL degraded percentage</b></td></tr>
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<tr><td class="row"><b>Measurement device</b></td><td><b>  HSL degraded percentage </b></td></tr>
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<tr><td class="row">BBa_K516220 </td><td class="row"> 96.11 [3.66]</td></tr>
 
<tr><td class="row">BBa_K516220 </td><td class="row"> 96.11 [3.66]</td></tr>
 
<tr><td class="row">BBa_K516221 </td><td class="row"> 77.60 [2.66]</td></tr>
 
<tr><td class="row">BBa_K516221 </td><td class="row"> 77.60 [2.66]</td></tr>
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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):
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In order to evaluate the RBSs efficiency, the ratio between the percentage of the degraded HSL realtive to the measurement device with the standard RBS (BBa_B0034) was computed:
  
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<div align="center">
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<table class="data" border="1">
 
<tr><td class="row"><b>RBS</b></td><td><b>  RBS efficiency</b></td></tr>
 
<tr><td class="row"><b>RBS</b></td><td><b>  RBS efficiency</b></td></tr>
 
<tr><td class="row">BBa_B0030 </td><td class="row"> 0.96</td></tr>
 
<tr><td class="row">BBa_B0030 </td><td class="row"> 0.96</td></tr>

Latest revision as of 21:31, 1 November 2017

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_C0060

User Reviews

UNIQ4a8c54aee3dd4e85-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:


A system of differential equations has been derived and parameters would have been identified, studying the exponential growth phase:
The four measurment parts ptet-RBSx-LuxI were quantitatively characetrized using the BBa_T9002 biosensor. Unfortunately the model parameters (kcat, kM,AiiA) were not estimated because placing the device in low copy plasmid (pSB4C5)resulted in no HSL degradation, as shown in the figures below:
A semi-quantitative information about AiiA enzyme activity is given, 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 a measure of AiiA activity is given, 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, the ratio between the percentage of the degraded HSL realtive to the measurement device with the standard RBS (BBa_B0034) was computed:
RBS RBS efficiency
BBa_B0030 0.96
BBa_B0031 0.78
BBa_B0032 0.51
BBa_B0034 1

UNIQ4a8c54aee3dd4e85-partinfo-00000003-QINU