Team:METU Turkey characterization

From 2014.igem.org

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<h3>Our project mainly depends on Catechol (1,2-benzenediol) which is a chemical that come off at PET degredation. </h3>
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<h1> Characterization of K1392991</h1>
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<h3>* At the following picture, there is Catechol (1,2-benzenediol) present in Left tube. It proves that in presence of catechol there is a degredation process in k316003+j23108. </h3>
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<h3> There is no Catechol (1,2-benzenediol) present in Right tube. It proves that in absence of catechol there is no degredation of PET in k316003+j23108. </h3>
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<h3> Here, we characterized the part K1392991 by ligasing K316003 and J23108. After centrifugation E. coli DH5a at 3000rpm for 3 minutes, we burst the cells and let the enzymes free in solution. Then, adding 1 mL of catechol(0.1M) into tube, color change into green. </h3>
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<h3> This characterization contains parts that Imperial_College_London created in year 2010, we have proved that it works well and we aproved its function with our own construct of its use. </h3>
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<p> At the following picture, there is Catechol (1,2-benzenediol) present in Left tube. It proves that in presence of catechol there is a degredation process in k316003+j23108. </h3>
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<p> There is no Catechol (1,2-benzenediol) present in Right tube. It proves that in absence of catechol there is no degredation of PET in k316003+j23108. </p>
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<p> This characterization contains parts that <a href="http://parts.igem.org/Part:BBa_K316003:Experience">Imperial_College_London</a> created in year 2010, we have proved that it works well and we aproved its function with our own construct of its use. </p>
<img src="https://static.igem.org/mediawiki/2014/c/cf/Karakter.jpg">
<img src="https://static.igem.org/mediawiki/2014/c/cf/Karakter.jpg">
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<h3>* At the following picture, the liquid is catechol (1,2 benzenediol) itself.</h3>
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<h3> At the following picture, the liquid is 0.1M catechol (1,2 benzenediol) itself:</h3>
<p>  <img src="https://static.igem.org/mediawiki/2014/6/6d/Catecholkoyduk.jpg"> </p>
<p>  <img src="https://static.igem.org/mediawiki/2014/6/6d/Catecholkoyduk.jpg"> </p>
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<img src="https://static.igem.org/mediawiki/2014/0/0a/DwikitemplateB_dnastrand.png">
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<img src="https://static.igem.org/mediawiki/2014/e/e9/Asilcircularlin.png">
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<h3> How does our project work?</h3>
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<p> Totally degradation of PET to pyruvate is done by E. coli. </p>
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<p> According to gel image: Left, part is closed to 2300bp. Right, linearized backbone is closed to 4000bp. </p>
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<img src="https://static.igem.org/mediawiki/2014/7/7d/Gears.png">
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<img src="https://static.igem.org/mediawiki/2014/e/e8/Hydratase.PNG">
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<h3> Who will our project help?</h3>
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<h3> K1392931 (Hydratase, 810bp)</h3>
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<p> Our project will help the environment to become cleaner.</p>
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<p> We got those photos after PCR of E. coli BL21 (DE3). We have tried colony and liquid culture of E coli. BL21 (DE3)</p>
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<img src="https://static.igem.org/mediawiki/2014/9/96/Camurpet.jpg">
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<img src="https://static.igem.org/mediawiki/2014/1/1c/Aldolase.PNG">
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<h3>Why did we choose this project?</h3>
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<h3>K1392932 (Aldolase, 1014bp)</h3>
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<p> The biodegradation of plastic bottles took thousands of years, and the degrading process emits toxic chemicals into the air. Our project aims to clean enviroment with the degredation of PET to pyruvate by E.coli. With this project,while  the environment will be cleaned from PET, the E.coli would add pyruvate to the cell cycle and use it as a new source. </p>
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<p> We got those photos after PCR of E. coli BL21 (DE3). We have tried colony and liquid culture of E coli. BL21 (DE3) </p>
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Latest revision as of 02:10, 18 October 2014

Team:METU Turkey/Templates/Navigationbar

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Characterization of K1392991

Here, we characterized the part K1392991 by ligasing K316003 and J23108. After centrifugation E. coli DH5a at 3000rpm for 3 minutes, we burst the cells and let the enzymes free in solution. Then, adding 1 mL of catechol(0.1M) into tube, color change into green.

At the following picture, there is Catechol (1,2-benzenediol) present in Left tube. It proves that in presence of catechol there is a degredation process in k316003+j23108.

There is no Catechol (1,2-benzenediol) present in Right tube. It proves that in absence of catechol there is no degredation of PET in k316003+j23108.

This characterization contains parts that Imperial_College_London created in year 2010, we have proved that it works well and we aproved its function with our own construct of its use.

At the following picture, the liquid is 0.1M catechol (1,2 benzenediol) itself:

According to gel image: Left, part is closed to 2300bp. Right, linearized backbone is closed to 4000bp.

K1392931 (Hydratase, 810bp)

We got those photos after PCR of E. coli BL21 (DE3). We have tried colony and liquid culture of E coli. BL21 (DE3)

K1392932 (Aldolase, 1014bp)

We got those photos after PCR of E. coli BL21 (DE3). We have tried colony and liquid culture of E coli. BL21 (DE3)

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