Difference between revisions of "Team:Vilnius-Lithuania/Demonstrate"
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| + | .content-block ul{ | ||
| + | list-style-image: url(https://2019.igem.org/wiki/images/e/ee/T--Vilnius-Lithuania--bullet.png)!important; | ||
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font-family: 'intro', sans-serif!important; | font-family: 'intro', sans-serif!important; | ||
margin: 0!important; | margin: 0!important; | ||
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| + | font-family: 'intro', sans-serif!important; | ||
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img{ | img{ | ||
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<a href="#">Project</a> | <a href="#">Project</a> | ||
<a href="https://2019.igem.org/Team:Vilnius-Lithuania/Description" class="sub">Inspiration & Description</a> | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Description" class="sub">Inspiration & Description</a> | ||
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<a href="https://2019.igem.org/Team:Vilnius-Lithuania/Transcriptional_Regulation" class="sub">Transcriptional regulation</a> | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Transcriptional_Regulation" class="sub">Transcriptional regulation</a> | ||
<a href="https://2019.igem.org/Team:Vilnius-Lithuania/Plasmid_Copy_Number_Control" class="sub">Plasmid copy number control</a> | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Plasmid_Copy_Number_Control" class="sub">Plasmid copy number control</a> | ||
| − | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Degradation_Rate_Control" class="sub"> | + | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Degradation_Rate_Control" class="sub">Post-translational regulation</a> |
<a href="https://2019.igem.org/Team:Vilnius-Lithuania/Demonstrate" class="sub">Demonstration</a> | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Demonstrate" class="sub">Demonstration</a> | ||
<a href="https://2019.igem.org/Team:Vilnius-Lithuania/Parts" class="sub">Parts</a> | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Parts" class="sub">Parts</a> | ||
<a href="#">Modeling</a> | <a href="#">Modeling</a> | ||
<a href="https://2019.igem.org/Team:Vilnius-Lithuania/Model" class="sub">Overview</a> | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Model" class="sub">Overview</a> | ||
| − | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Degradation_Rate_Control_Model" class="sub"> | + | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Degradation_Rate_Control_Model" class="sub">Post-translational regulation</a> |
| − | + | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Transcriptional_Regulation_CarH" class="sub">Transcriptional regulation (CarH)</a> | |
| − | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Transcriptional_Regulation_CarH" class="sub">Transcriptional | + | |
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<a href="https://2019.igem.org/Team:Vilnius-Lithuania/Software">Software</a> | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Software">Software</a> | ||
<a href="#">Human Practices</a> | <a href="#">Human Practices</a> | ||
| − | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Human_Practices" class="sub">Human Practices</a> | + | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Human_Practices" class="sub">Integrated Human Practices</a> |
<a href="https://2019.igem.org/Team:Vilnius-Lithuania/Public_Engagement" class="sub">Education & Public Engagment</a> | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Public_Engagement" class="sub">Education & Public Engagment</a> | ||
<a href="https://2019.igem.org/Team:Vilnius-Lithuania/Collaborations" class="sub">Collaboration</a> | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Collaborations" class="sub">Collaboration</a> | ||
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<ul class="menu sub-menu-project sub-menu"> | <ul class="menu sub-menu-project sub-menu"> | ||
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<a href="#" class="return return-project" onclick="returnMenu('project')">Back to Main Menu</a> | <a href="#" class="return return-project" onclick="returnMenu('project')">Back to Main Menu</a> | ||
</li> | </li> | ||
| − | <li class="menu-item" style="position: relative; | + | <li class="menu-item" style="position: relative;"> |
<a href="https://2019.igem.org/Team:Vilnius-Lithuania/Description">Inspiration & Description</a> | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Description">Inspiration & Description</a> | ||
</li> | </li> | ||
| − | <li class="menu-item" style="position: relative; | + | <li class="menu-item" style="position: relative;"> |
| − | <a href=" | + | <a href="#">Design & Results:</a> |
</li> | </li> | ||
| − | <li class="menu-item" style="position: relative | + | <li class="menu-item menu-item-bullet" style="position: relative;padding-left:24px"> |
| − | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Transcriptional_Regulation">Transcriptional regulation</a> | + | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Transcriptional_Regulation">Transcriptional regulation</a> |
</li> | </li> | ||
| − | <li class="menu-item" style="position: relative | + | <li class="menu-item menu-item-bullet" style="position: relative;padding-left:24px"> |
<a href="https://2019.igem.org/Team:Vilnius-Lithuania/Plasmid_Copy_Number_Control">Plasmid copy number control</a> | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Plasmid_Copy_Number_Control">Plasmid copy number control</a> | ||
</li> | </li> | ||
| − | <li class="menu-item" style="position: relative | + | <li class="menu-item menu-item-bullet" style="position: relative;padding-left:24px"> |
| − | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Degradation_Rate_Control"> | + | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Degradation_Rate_Control">Post-translational regulation</a> |
</li> | </li> | ||
| − | <li class="menu-item" style="position: relative; | + | <li class="menu-item" style="position: relative;"> |
| − | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Demonstrate">Demonstration</a> | + | <a class="active" href="https://2019.igem.org/Team:Vilnius-Lithuania/Demonstrate">Demonstration</a> |
</li> | </li> | ||
| − | <li class="menu-item" style="position: relative; | + | <li class="menu-item" style="position: relative;"> |
<a href="https://2019.igem.org/Team:Vilnius-Lithuania/Parts">Parts</a> | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Parts">Parts</a> | ||
| + | </li> | ||
| + | <li class="menu-item" style="position: relative;"> | ||
| + | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Measurement">Measurement</a> | ||
</li> | </li> | ||
</ul> | </ul> | ||
| − | <ul class="menu sub-menu-modeling sub-menu"> | + | <ul class="menu sub-menu-modeling sub-menu" style="visibility:hidden"> |
<li class="menu-item" style="position: relative;left:200px;opacity:0"> | <li class="menu-item" style="position: relative;left:200px;opacity:0"> | ||
<a href="#" class="return" onclick="returnMenu('modeling')">Back to Main Menu</a> | <a href="#" class="return" onclick="returnMenu('modeling')">Back to Main Menu</a> | ||
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</li> | </li> | ||
<li class="menu-item" style="position: relative;left:200px;opacity:0"> | <li class="menu-item" style="position: relative;left:200px;opacity:0"> | ||
| − | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Degradation_Rate_Control_Model"> | + | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Degradation_Rate_Control_Model">Post-translational regulation</a> |
</li> | </li> | ||
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<li class="menu-item" style="position: relative;left:200px;opacity:0"> | <li class="menu-item" style="position: relative;left:200px;opacity:0"> | ||
| − | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/ | + | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Transcriptional_Regulation_CarH">Transcriptional regulation (CarH)</a> |
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</li> | </li> | ||
</ul> | </ul> | ||
| − | <ul class="menu sub-menu-software sub-menu"> | + | <ul class="menu sub-menu-software sub-menu" style="visibility:hidden"> |
<li class="menu-item" style="position: relative;left:200px;opacity:0"> | <li class="menu-item" style="position: relative;left:200px;opacity:0"> | ||
<a href="#" class="return" onclick="returnMenu('software')">Back to Main Menu</a> | <a href="#" class="return" onclick="returnMenu('software')">Back to Main Menu</a> | ||
</li> | </li> | ||
</ul> | </ul> | ||
| − | <ul class="menu sub-menu-hp sub-menu"> | + | <ul class="menu sub-menu-hp sub-menu" style="visibility:hidden"> |
<li class="menu-item" style="position: relative;left:200px;opacity:0"> | <li class="menu-item" style="position: relative;left:200px;opacity:0"> | ||
<a href="#" class="return" onclick="returnMenu('hp')">Back to Main Menu</a> | <a href="#" class="return" onclick="returnMenu('hp')">Back to Main Menu</a> | ||
</li> | </li> | ||
| − | <li class="menu-item" style="position: relative;left:200px;opacity:0"> | + | <li class="menu-item" style="position: relative;left:200px;opacity:0;white-space: nowrap"> |
| − | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Human_Practices">Human Practices</a> | + | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Human_Practices">Integrated Human Practices</a> |
</li> | </li> | ||
<li class="menu-item" style="position: relative;left:200px;opacity:0"> | <li class="menu-item" style="position: relative;left:200px;opacity:0"> | ||
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</ul> | </ul> | ||
| − | <ul class="menu sub-menu-team sub-menu"> | + | <ul class="menu sub-menu-team sub-menu" style="visibility:hidden"> |
<li class="menu-item" style="position: relative;left:200px;opacity:0"> | <li class="menu-item" style="position: relative;left:200px;opacity:0"> | ||
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| − | <ul class="menu sub-menu-notebook sub-menu"> | + | <ul class="menu sub-menu-notebook sub-menu" style="visibility:hidden"> |
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<a href="https://2019.igem.org/Team:Vilnius-Lithuania">Home</a> | <a href="https://2019.igem.org/Team:Vilnius-Lithuania">Home</a> | ||
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<a href="#">Project</a> | <a href="#">Project</a> | ||
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<a href="#">Modeling</a> | <a href="#">Modeling</a> | ||
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<a href="https://2019.igem.org/Team:Vilnius-Lithuania/Software">Software</a> | <a href="https://2019.igem.org/Team:Vilnius-Lithuania/Software">Software</a> | ||
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<a href="#">Human Practices</a> | <a href="#">Human Practices</a> | ||
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<a href="#">Team</a> | <a href="#">Team</a> | ||
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<a href="#">Notebook</a> | <a href="#">Notebook</a> | ||
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<div class="cover"> | <div class="cover"> | ||
| − | <img src="https://2019.igem.org/wiki/images/ | + | <img src="https://2019.igem.org/wiki/images/9/98/T--Vilnius-Lithuania--cover13.jpg"> |
| − | <span> | + | <span>Demonstration</span> |
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| − | + | <p>The proof of Colight consist of several critical steps. </p><br> | |
| − | + | ||
| − | + | <h4 class="page-heading">1.The proof that CarH works as light inducible repressor</h4><br> | |
| − | + | ||
| − | + | <img src="https://2019.igem.org/wiki/images/c/c6/T--Vilnius-Lithuania--Trans-Carh1.png" style="width:100%"><br><br> | |
| − | + | ||
| − | + | <p>The experiment shows that CarH expressing cells grown in the dark for 12 hours have a substantially reduced level of fluorescence. That is because CarH represses its promoter under dark conditions and the promoter is induced under light.</p><br> | |
| + | |||
| + | <h4 class="page-heading">2.The proof of light inducible protein degradation by mf-Lon</h4><br> | ||
| + | |||
| + | <img src="https://2019.igem.org/wiki/images/e/e3/T--Vilnius-Lithuania--Degradation-2.png" style="width:100%"><br><br> | ||
| + | |||
| + | <p>In this graph we see that a protein of a construct number 160 has its protein level reduced after induction by light. That is because after induction asLOV change its conformation, and degradation tag becomes accessible to mf-Lon protease which degrades sfGFP.</p><br> | ||
| + | |||
| + | <h4 class="page-heading">3.The proof of light modular plasmid copy number regulation</h4><br> | ||
| + | |||
| + | |||
| + | <img src="https://2019.igem.org/wiki/images/a/a1/T--Vilnius-Lithuania--copy2.png" style="width:100%"><br><br> | ||
| + | |||
| + | <p>In this graph we show that illuminated cells, compared to uniluminated, have their plasmid copy number increased (grown until 16 hours). And when the light is turned off for both bacteria culture samples, the plasmid copy number bacteria is equalized.</p><br> | ||
| + | |||
| + | |||
| + | |||
| + | |||
</div> | </div> | ||
</div> | </div> | ||
Latest revision as of 03:46, 22 October 2019
Demonstration
The proof of Colight consist of several critical steps.
1.The proof that CarH works as light inducible repressor
The experiment shows that CarH expressing cells grown in the dark for 12 hours have a substantially reduced level of fluorescence. That is because CarH represses its promoter under dark conditions and the promoter is induced under light.
2.The proof of light inducible protein degradation by mf-Lon
In this graph we see that a protein of a construct number 160 has its protein level reduced after induction by light. That is because after induction asLOV change its conformation, and degradation tag becomes accessible to mf-Lon protease which degrades sfGFP.
3.The proof of light modular plasmid copy number regulation
In this graph we show that illuminated cells, compared to uniluminated, have their plasmid copy number increased (grown until 16 hours). And when the light is turned off for both bacteria culture samples, the plasmid copy number bacteria is equalized.