Line 462: | Line 462: | ||
<li><a href="#section2">Recycling part</a></li> | <li><a href="#section2">Recycling part</a></li> | ||
<li><a href="#section3">The whole circuit </a></li> | <li><a href="#section3">The whole circuit </a></li> | ||
+ | <li><a href="#section4">Detection of <br>sensing part</a></li> | ||
+ | <li><a href="#section5">Detection of adsorption<br>capacity of LBTs </a></li> | ||
</ul> | </ul> | ||
</div> | </div> | ||
Line 583: | Line 585: | ||
<div id="section2" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;"> | <div id="section2" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;"> | ||
<h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">Recycling part </h3> | <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">Recycling part </h3> | ||
− | <p> | + | <p><strong>The recycling part is to capture the rare earth ions in the environment and to anchor our cells over the silicon net.</strong></p> |
− | + | ||
− | + | ||
<h4><strong>①With appropriate primers, PCR was carried out to amplify target gene oprf-LBT1-12 and oprf-Si tag.</strong> </h4> | <h4><strong>①With appropriate primers, PCR was carried out to amplify target gene oprf-LBT1-12 and oprf-Si tag.</strong> </h4> | ||
− | |||
− | |||
− | |||
<h4 class="col-xs-12"><strong>②Then we constructed two vectors to achieve expression of LBT and Si-tag.</strong> </h4> | <h4 class="col-xs-12"><strong>②Then we constructed two vectors to achieve expression of LBT and Si-tag.</strong> </h4> | ||
− | + | <img title="demo1" src="https://static.igem.org/mediawiki/2017/7/73/2017_HUST_China_Experiment_image2.png" alt="demo1" class="col-xs-4 col-xs-offset-1" style="padding: 10px 0px;"> | |
− | + | <img title="demo1" src="https://static.igem.org/mediawiki/2017/7/78/2017_HUST_China_Experiment_image3.png" alt="experiment_circuit" alt="demo1" class="col-xs-4 col-xs-offset-2" style="padding: 10px 0px;"> | |
− | + | ||
+ | |||
<p class="col-xs-12"> | <p class="col-xs-12"> | ||
We cloned the sequence of oprf-LBT and Si-tag into the vector pET28α in order to make the sequence be tagged N-terminally with a 6×His tag. 6×His tag is an affinity tag which allows the tagged recombinant protein to be purified in the process of affinity purification. | We cloned the sequence of oprf-LBT and Si-tag into the vector pET28α in order to make the sequence be tagged N-terminally with a 6×His tag. 6×His tag is an affinity tag which allows the tagged recombinant protein to be purified in the process of affinity purification. | ||
Line 608: | Line 606: | ||
All of our part should be built in one plasmid. Therefore, we use endonuclease and ligase to replace the GFP by Oprf-LBT and Oprf-Si tag. | All of our part should be built in one plasmid. Therefore, we use endonuclease and ligase to replace the GFP by Oprf-LBT and Oprf-Si tag. | ||
</p> | </p> | ||
− | + | <img title="demo1" src="https://static.igem.org/mediawiki/2017/5/58/2017_HUST_China_Experiment_image4.png" alt="experiment_circuit" alt="demo1" class="col-xs-4 col-xs-offset-4" style="padding: 10px 0px;"> | |
− | + | ||
− | + | ||
</div> | </div> | ||
+ | |||
+ | <div id="section4" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;"> | ||
+ | <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px;margin-top:0;">Detection of sensing part </h3> | ||
+ | <p>We use our medium containing a small amount of rare earth ions to culture our E. coli. When our engineered E. coli senses the existence of rare earth ions, the pmrC promoter will activate the transcription of the GFP gene. Through the detection of GFP expression, we can know the efficacy of the sensing part. | ||
+ | </p> | ||
+ | </div> | ||
+ | |||
+ | <div id="section5" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;"> | ||
+ | <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px;margin-top:0;">Detection of adsorption capacity of LBTs </h3> | ||
+ | <p class="col-xs-12">A method of precipitation titration is used to detect the level of rare earth ions adsorption. We add a certain concentration of bacteria to the solution containing different concentrations of rare earth ions. Then centrifuge to remove the bacteria in the solution. | ||
+ | </p> | ||
+ | |||
+ | <img title="demo1" src="https://static.igem.org/mediawiki/2017/1/1a/2017_HUST_China_Experiment_image5.png" alt="experiment_circuit" alt="demo1" class="col-xs-4 col-xs-offset-4" style="padding: 10px 0px;"> | ||
+ | |||
+ | <p class="col-xs-12">In order to make the terbium ions sufficiently precipitated, we add excess base. Rinse the precipitate and dissolve the precipitate with hydrochloric acid titration. In this way, we can know the amount of terbium ions which have already adsorbed on the surface of the bacteria. | ||
+ | </p> | ||
+ | |||
+ | <img title="demo1" src="https://static.igem.org/mediawiki/2017/f/fc/2017_HUST_China_Experiment_image6.png" alt="experiment_circuit" alt="demo1" class="col-xs-8 col-xs-offset-2" style="padding: 10px 0px;"> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | |||
+ | |||
</div> | </div> | ||
+ | |||
<nav class="col-sm-2" id="myScrollspy2.0"> | <nav class="col-sm-2" id="myScrollspy2.0"> |
Revision as of 12:19, 30 October 2017