Difference between revisions of "Part:BBa C0061:Experience"

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NB: unless differently specified, all tests were performed in <a href='http://2011.igem.org/Team:UNIPV-Pavia/Protocols#MG1655Z1'><em>E. coli</em> MGZ1</a> in M9 supplemented medium at 37°C in low copy plasmid <A HREF="https://parts.igem.org/wiki/index.php/Part:pSB4C5">pSB4C5</a>.
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The LuxI enzyme activity has been characterized under the regulation of p<sub>tet</sub> promoter, assaying its enzymatic activity, with these parts:
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<li> <A HREF="https://parts.igem.org/wiki/index.php/Part: BBa_K516210 "> BBa_K516210 </a> (wiki name: E13 ) pTet-RBS30-LuxI </li>
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<li> <A HREF="https://parts.igem.org/wiki/index.php/Part: BBa_K516211 "> BBa_K516211 </a> (wiki name: E14 ) pTet-RBS31-LuxI </li>
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<li> <A HREF="https://parts.igem.org/wiki/index.php/Part: BBa_K516212 "> BBa_K516212 </a> (wiki name: E15 ) pTet-RBS32-LuxI </li>
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<li> <A HREF="https://parts.igem.org/wiki/index.php/Part: BBa_K516214 "> BBa_K516214 </a> (wiki name: E16 ) pTet-RBS34-LuxI </li></ul>
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<em>Though these parts don't have a transcriptional terminator, they have been characterized in low copy plasmid <A HREF="https://parts.igem.org/wiki/index.php/Part:pSB4C5">pSB4C5</a>, that contains the BBa_B0054 terminator.</em>
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The four measurment parts <a href="http://2011.igem.org/Team:UNIPV-Pavia/Parts/Characterized#pTetLuxI">p<sub>tet</sub>-RBSx-LuxI</a> were quantitatively characetrized using the BBa_T9002 biosensor see <a href='http://2011.igem.org/Team:UNIPV-Pavia/Project/Modelling#introduction_to_T9002'>modelling section</a>.<br>
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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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A significative HSL production was observed for <A HREF="https://parts.igem.org/wiki/index.php/Part:BBa_K516210">BBa_K516210</a>, <A HREF="https://parts.igem.org/wiki/index.php/Part:BBa_K516211">BBa_K516211</a> and <A HREF="https://parts.igem.org/wiki/index.php/Part:BBa_K516214">BBa_K516214</a>, while no HSL production was observed for <A HREF="https://parts.igem.orgwiki/index.php/Part:BBa_K516212">BBa_K516212</a>. 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>:
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<td class='row'><b>V<sub>max</sub></b></td>
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<td class='row'><b>k<sub>M,LuxI</sub></b></td>
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<td class='row'><b>&alpha;<sub>B0030</sub></b></td>
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<td class='row'><b>&alpha;<sub>B0031</sub></b></td>
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<td class='row'><b>&alpha;<sub>B0032</sub></b></td>
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<td class='row'><b>&alpha;<sub>B0034</sub></b></td>
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</tr>
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<tr><td class='row'>3.56*10<sup>-9</sup></td>
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<td class='row'>6.87*10<sup>3</sup></td>
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<td class='row'>87</td>
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<td class='row'>8.5</td>
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<td class='row'>ND</td>
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<td class='row'>252</td>
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</tr></table>
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<!-- DON'T DELETE --><partinfo>BBa_C0061 EndReviews</partinfo>
 
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Revision as of 11:02, 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_C0061

User Reviews

UNIQ94e6ff7097f3800b-partinfo-00000000-QINU

Antiquity

This review comes from the old result system and indicates that this part did not work in some test.

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wmholtz

Using this part, I have successfully constructed and tested a quorum sensing circuit in E. coli.

•••••

Youri

This part was used as a subpart in K546005, K546546 and found to be functional.

;

UNIQ94e6ff7097f3800b-partinfo-00000004-QINU



UNIQ94e6ff7097f3800b-partinfo-00000005-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 LuxI enzyme activity has been characterized under the regulation of ptet promoter, assaying its enzymatic activity, with these parts:



Though these parts don't have a transcriptional terminator, they have been characterized in low copy plasmid pSB4C5, that contains the BBa_B0054 terminator. The four measurment parts ptet-RBSx-LuxI were quantitatively characetrized using the BBa_T9002 biosensor see modelling section.
The HSL synthesis rate has been evaluated according to the model equations, properly adjusted.
A significative HSL production was observed for BBa_K516210, BBa_K516211 and BBa_K516214, while no HSL production was observed for BBa_K516212. The parameters Vmax, kM,LuxI and αRBSx were estimated with a simultaneous fitting of the data collected as described in measurement section:
Vmax kM,LuxI αB0030 αB0031 αB0032 αB0034
3.56*10-9 6.87*103 87 8.5 ND 252

UNIQ94e6ff7097f3800b-partinfo-00000008-QINU