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| {{HSNU-TAIPEI/main}} | | {{HSNU-TAIPEI/main}} |
| <html> | | <html> |
| + | <style> |
| + | ul, ol { |
| + | margin: 0 !important; |
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| <div class="wrapper"> | | <div class="wrapper"> |
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| <a>Project</a> | | <a>Project</a> |
| <ul class="sub-nav"> | | <ul class="sub-nav"> |
− | <li class="list-item"><a href="https://2015.igem.org/Team:HSNU-TAIPEI/projectoverview">Overview</a></li> | + | <li class="list-item"><a href="https://2015.igem.org/Team:HSNU-TAIPEI/Description">Overview</a></li> |
| <li class="list-item"><a href="https://2015.igem.org/Team:HSNU-TAIPEI/projectBP">Benzo[A]Pyrene</a></li> | | <li class="list-item"><a href="https://2015.igem.org/Team:HSNU-TAIPEI/projectBP">Benzo[A]Pyrene</a></li> |
− | <li class="list-item"><a href="https://2015.igem.org/Team:HSNU-TAIPEI/projectcopper">Copper</a></li>
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| <li class="list-item"><a href="https://2015.igem.org/Team:HSNU-TAIPEI/projectcadmium">Cadmium</a></li> | | <li class="list-item"><a href="https://2015.igem.org/Team:HSNU-TAIPEI/projectcadmium">Cadmium</a></li> |
− | <li class="list-item"><a href="https://2015.igem.org/Team:HSNU-TAIPEI/projectmercury">Mercury</a></li>
| |
| <li class="list-item"><a href="https://2015.igem.org/Team:HSNU-TAIPEI/projectlead">Lead</a></li> | | <li class="list-item"><a href="https://2015.igem.org/Team:HSNU-TAIPEI/projectlead">Lead</a></li> |
| <li class="list-item"><a href="https://2015.igem.org/Team:HSNU-TAIPEI/projectafla">Aflatoxin</a></li> | | <li class="list-item"><a href="https://2015.igem.org/Team:HSNU-TAIPEI/projectafla">Aflatoxin</a></li> |
| </ul> | | </ul> |
| </li> | | </li> |
− | <li class="list-item"><a href="https://2015.igem.org/Team:HSNU-TAIPEI/product">Product</a></li> | + | <li class="list-item"><a href="https://2015.igem.org/Team:HSNU-TAIPEI/Design">Product</a></li> |
− | <li class="list-item"><a href="https://2015.igem.org/Team:HSNU-TAIPEI/parts">Parts</a></li> | + | <li class="list-item has-sub-nav"> |
| + | <a>Parts</a> |
| + | <ul class="sub-nav"> |
| + | <li class="list-item"><a href="https://2015.igem.org/Team:HSNU-TAIPEI/Parts">Parts</a></li> |
| + | <li class="list-item"><a href="https://2015.igem.org/Team:HSNU-TAIPEI/Basic_Part">Basic Part</a></li> |
| + | <li class="list-item"><a href="https://2015.igem.org/Team:HSNU-TAIPEI/Composite_Part">Composite Part</a></li> |
| + | <li class="list-item"><a href="https://2015.igem.org/Team:HSNU-TAIPEI/Part_Collection">Part Collection</a></li> |
| + | </ul> |
| + | </li> |
| <li class="list-item"><a href="https://2015.igem.org/Team:HSNU-TAIPEI/labnotes">Notebook</a></li> | | <li class="list-item"><a href="https://2015.igem.org/Team:HSNU-TAIPEI/labnotes">Notebook</a></li> |
| <li class="list-item has-sub-nav"> | | <li class="list-item has-sub-nav"> |
| <a>Human Practice</a> | | <a>Human Practice</a> |
| <ul class="sub-nav"> | | <ul class="sub-nav"> |
− | <li class="list-item"><a href="https://2015.igem.org/Team:HSNU-TAIPEI/hp/overview">Overview</a></li> | + | <li class="list-item"><a href="https://2015.igem.org/Team:HSNU-TAIPEI/Practices">Overview</a></li> |
| <li class="list-item has-sub-nav"> | | <li class="list-item has-sub-nav"> |
| <a>Organization</a> | | <a>Organization</a> |
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| <main> | | <main> |
| <div class="mc-container"> | | <div class="mc-container"> |
− | | + | <h1>Project<span><em>Benzo[A]Pyrene</em></span></h1> |
− | <h1>Project<span><em>Benzo[A]Pyrene</em></span></h1> | + | |
| <div class="section"> | | <div class="section"> |
| <article class="article"> | | <article class="article"> |
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| <li> | | <li> |
| <span>Why do we detect benzo[a]pyrene?</span> | | <span>Why do we detect benzo[a]pyrene?</span> |
− | <p class="article-p">Because when food,such as fried food ,which is heated above 300 degrees celcius,food will release benzo[a]pyrene recycled oil usually contain benzo[a]pyrene[1].</p> | + | <p class="article-p">Because when food, such as fried food , is heated above 300 degrees celcius, benzo[a]pyrene is released. That is why recycled oil usually contains benzo[a]pyrene [1].</p> |
| </li> | | </li> |
| <li> | | <li> |
| <span>The harm of benzo[a]pyrene</span> | | <span>The harm of benzo[a]pyrene</span> |
− | <p class="article-p">Pathway:from skin,breathing in,eating Carcinogenic,environmental pollution, Skin irritation,eye irritation[1].</p> | + | <p class="article-p">Pathway: from skin, breathing in, and eating. It is carcinogenic and causes environmental pollution. It causes skin irritation and eye irritation [1].</p> |
| </li> | | </li> |
| <li> | | <li> |
| <span>Taiwanese regulation</span> | | <span>Taiwanese regulation</span> |
| + | <p class="article-p table-note">▼Table1:The regulation of Benzo[a]pyrene in Taiwan.</p> |
| <div class="article-img"><img src="https://static.igem.org/mediawiki/2015/9/94/2015hsnu-benzo-a-pyrene_5.png"></div> | | <div class="article-img"><img src="https://static.igem.org/mediawiki/2015/9/94/2015hsnu-benzo-a-pyrene_5.png"></div> |
| </li> | | </li> |
| <li> | | <li> |
| <span>National regulation</span> | | <span>National regulation</span> |
− | <ul class="article-ul">
| |
− | <li>Europe:The same to Taiwanese regulation[2]</li>
| |
− | <li>American:The MCL has been set at 0.2 ppb[3]</li>
| |
− | </ul>
| |
| </li> | | </li> |
| + | <ul class="article-ul"> |
| + | <li>Europe:same to Taiwanese regulation[2]</li> |
| + | <li>American:The MCL has been set at 0.2 ppb[3]</li> |
| + | </ul> |
| </ol> | | </ol> |
| </article> | | </article> |
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| <p class="article-p">So far,we don’t find the protein which can combine with the Benzo[a]pyrene,so first we must degradate it by Laccase. And its product is Bap-1,6-quinone which can combine with the protein,in order that we can detect Benzo[a]pyrene.[4] We put the gene fragment which can produce the Laccase in the E.coli. Let it produce itself.</p> | | <p class="article-p">So far,we don’t find the protein which can combine with the Benzo[a]pyrene,so first we must degradate it by Laccase. And its product is Bap-1,6-quinone which can combine with the protein,in order that we can detect Benzo[a]pyrene.[4] We put the gene fragment which can produce the Laccase in the E.coli. Let it produce itself.</p> |
| <div class="article-img"><img src="https://static.igem.org/mediawiki/2015/e/ed/2015hsnu-benzo-a-pyrene_2.png"></div> | | <div class="article-img"><img src="https://static.igem.org/mediawiki/2015/e/ed/2015hsnu-benzo-a-pyrene_2.png"></div> |
| + | <p class="article-p">▲Fig.1-1:The circuit of detecting Benzo[a]pyrene.</p> |
| <p class="article-p">QsrR is a protein which can combine with Bap-1,6-quinone.[5]As Laccase,we put the gene fragment in the E.coli. Let it produce itself.</p> | | <p class="article-p">QsrR is a protein which can combine with Bap-1,6-quinone.[5]As Laccase,we put the gene fragment in the E.coli. Let it produce itself.</p> |
| <div class="article-img"><img src="https://static.igem.org/mediawiki/2015/7/7d/2015hsnu-benzo-a-pyrene_3.png"></div> | | <div class="article-img"><img src="https://static.igem.org/mediawiki/2015/7/7d/2015hsnu-benzo-a-pyrene_3.png"></div> |
| + | <p class="article-p">▲Fig.1-2:The circuit of detecting Benzo[a]pyrene.</p> |
| <p class="article-p">We design a gene part make QsrR repress sfRFP’s production. Usually,QsrR is on the Binding Site,and red fluorescent protein will not be produced. When QsrR combine with Bap-1,6-quinone, QsrR will be activated and go away. And the transcription will carry on,then it will produce red fluorescent protein.</p> | | <p class="article-p">We design a gene part make QsrR repress sfRFP’s production. Usually,QsrR is on the Binding Site,and red fluorescent protein will not be produced. When QsrR combine with Bap-1,6-quinone, QsrR will be activated and go away. And the transcription will carry on,then it will produce red fluorescent protein.</p> |
| <div class="article-img"><img src="https://static.igem.org/mediawiki/2015/8/8e/2015hsnu-benzo-a-pyrene_4.png"></div> | | <div class="article-img"><img src="https://static.igem.org/mediawiki/2015/8/8e/2015hsnu-benzo-a-pyrene_4.png"></div> |
| + | <p class="article-p">▲Fig.1-3:The circuit of detecting Benzo[a]pyrene.</p> |
| + | </article> |
| + | <article class="article"> |
| + | <h3 class="article-title">Result</h3> |
| + | <p>Whether e.coli is alive in the poisons, condition or not</p> |
| + | <ol type="A"> |
| + | <li>Method</li> |
| + | <h3>DH5α-Pretest</h3> |
| + | <h4>Procedure</h4> |
| + | <p>Because we must test E.coli’s Survival in the environment there is benzo[a]pryene by counting the colonies,First we test how much concentration is the best.</p> |
| + | <ol> |
| + | <li> |
| + | <p>culture</p> |
| + | <p>STEP1:take 1μL DH5α to spread the plate(no Antibiotic)</p> |
| + | <p>STEP2:put in 37 degree Celsius 12~16hr</p> |
| + | </li> |
| + | <li> |
| + | <p>liquid culture</p> |
| + | </li> |
| + | <li> |
| + | <p>STEP1:put 80μL into 2ml LB broth </p> |
| + | <p>STEP2:recovering</p> |
| + | <p>STEP3: After 2hr,dilute it to 10<sup>-4</sup>,10<sup>-5</sup>,10<sup>-6</sup>,10<sup>-7</sup>,and then go to spread the plate (no Antibiotic)</p> |
| + | <p>STEP4: After 4hr dilute it to 10<sup>-4</sup>, 10<sup>-5</sup> ,10<sup>-6</sup> ,10<sup>-7</sup> ,and then go to spread the plate (no Antibiotic), 6hr and 8hr Similarly</p> |
| + | <p>STEP5:Take 200μL out from the tube and spread the plate(AMP+)</p> |
| + | <p>STEP6: put in 37 degree Celsius 12~16hr</p> |
| + | </li> |
| + | </ol> |
| + | <h3>Survival</h3> |
| + | <h4>Procedure</h4> |
| + | <p>First we culture DH5α with LB only plate for 15hr. Then,pick one colony in the LB broth,and liquid culture for 15hr. We divided two categories A and B.</p> |
| + | <h5>A:</h5> |
| + | <ol> |
| + | <li>Take 80μL into 2ml LB broth × 6 tubes and then culture 1 hr.</li> |
| + | <li>After 1hr,add 20μL benzo[a]pryene into three tubes(conc. Is 2000ppb(A thousand times the standard value))</li> |
| + | <li>And add 20μL DMSO into the other tubes.Then,culture for 3hr.</li> |
| + | <li>After 3hr,dilute the broth to 10<sup>-6</li> |
| + | <li>And take 200μL to spread the plate.</li> |
| + | </ol> |
| + | <h5>B:</h5> |
| + | <ol> |
| + | <li>Take 80μL into 2ml LB broth in a tube And then culture 1 hr.</li> |
| + | <li>After 1hr, put them into 6 tubes equally.</li> |
| + | <li>Dilute the broth to 5×10<sup>-4</sup></li> |
| + | <li>Add 0.4μL benzo[a]pryene(2×10<sup>-4</sup>) in three tubes.</li> |
| + | <li>Add 0.4μL DMSO in the other three tubes.</li> |
| + | <li>Go to 37 degree Celsius shaking for 10min.</li> |
| + | <li>Take 200μL to spread the plate.</li> |
| + | </ol> |
| + | <figure> |
| + | <figcaption>▼Table2: E. coli on the agar plate.</figcaption> |
| + | <img src="https://static.igem.org/mediawiki/2015/e/ee/HSNU-TAIPEI-BZP-904-1.jpg" width="70%"> |
| + | </figure> |
| + | <li>Results</li> |
| + | <figure> |
| + | <figcaption>▼Table3: E. coli on the agar plate.</figcaption> |
| + | <img src="https://static.igem.org/mediawiki/2015/2/2c/HSNU-TAIPEI-BZP-820-4.jpg"> |
| + | </figure> |
| + | <p> |
| + | The number of the colonies in the AMP+ plate is zero.<br> |
| + | According to the result, 2hr 10-5 and 4hr 10-6 is the best. |
| + | </p> |
| + | <figure> |
| + | <figcaption>▼Table4:Line Chart of Table 3.</figcaption> |
| + | <img src="https://static.igem.org/mediawiki/2015/b/bc/HSNU-TAIPEI-BZP-result.jpg"> |
| + | </figure> |
| + | <figure> |
| + | <img src="https://static.igem.org/mediawiki/2015/6/6b/HSNU-TAIPEI-BZP-820-1.jpg" width="50%"> |
| + | <figcaption>▲Fig.2:2hr agar only plate from left to rights are10<sup>-4</sup>, 10<sup>-5</sup>, 10<sup>-6</sup>, 10<sup>-7</sup>.</figcaption> |
| + | </figure> |
| + | <figure> |
| + | <img src="https://static.igem.org/mediawiki/2015/e/ef/HSNU-TAIPEI-BZP-820-2.jpg" width="50%"> |
| + | <figcaption>▲Fig3: 4hr agar only plate from left to rights are10<sup>-4</sup>, 10<sup>-5</sup>, 10<sup>-6</sup>, 10<sup>-7</sup>.</p> |
| + | <p class="note-caption">4hr plate from left to right is 10<sup>-4</sup>,10<sup>-5</sup>,10<sup>-6</sup>,10<sup>-7</sup></figcaption> |
| + | </figure> |
| + | <figure> |
| + | <img src="https://static.igem.org/mediawiki/2015/f/f7/HSNU-TAIPEI-BZP-820-3.jpg" width="50%"> |
| + | <figcaption>▲Fig4:Ampicillin Plate.</figcaption> |
| + | </figure> |
| + | <p>AMP+ Plate</p> |
| + | <p>According to the result, Beno[a]pryene does not affect E.coli’s survival.</p> |
| + | <p>But Category B is failed because its number of colony is too much.</p> |
| + | <figure> |
| + | <img src="https://static.igem.org/mediawiki/2015/d/d3/HSNU-TAIPEI-BZP-904-2.jpg" width="50%"> |
| + | <figcaption>▲Fig5: Benzo[a]pyrene Category A</figcaption> |
| + | </figure> |
| + | <p>Benzo[a]pryene Category A</p> |
| + | <figure> |
| + | <img src="https://static.igem.org/mediawiki/2015/6/66/HSNU-TAIPEI-BZP-904-3.jpg" width="50%"> |
| + | <figcaption>▲Fig6:Control Category A</figcaption> |
| + | </figure> |
| + | <p>Control Category A</p> |
| + | <figure> |
| + | <img src="https://static.igem.org/mediawiki/2015/6/69/HSNU-TAIPEI-BZP-904-4.jpg" width="50%"> |
| + | <figcaption></figcaption> |
| + | </figure> |
| + | <p>▲Fig7: Benzo[a]pyrene Category B</p> |
| + | <p>Benzo[a]pryene CategoryB</p> |
| + | <figure> |
| + | <img src="https://static.igem.org/mediawiki/2015/c/ce/HSNU-TAIPEI-BZP-904-5.jpg" width="50%"> |
| + | <figcaption>▲Fig8: Control Category B</figcaption> |
| + | </figure> |
| + | <p>Control CategoryB</p> |
| + | </ol> |
| </article> | | </article> |
− | <article class="article">
| |
− | <h3 class="article-title">Result</h3>
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− | <ol class="article-ol">
| |
− | <li>
| |
− | <span>Whether benzo[a]pyrene can enter e.coli or not</span>
| |
− | <ol class="article-ol" type="A">
| |
− | <li><span>Method</span></li>
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− | <li><span>Result</span></li>
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− | <li><span>Discussion</span></li>
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− | </ol>
| |
− | </li>
| |
− | <li>
| |
− | <span>Whether e.coli is alive in the poisons, condition or not</span>
| |
− | <ol class="article-ol" type="A">
| |
− | <li><span>Method</span></li>
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− | <li><span>Results</span></li>
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− | <li><span>Discussion</span></li>
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− | </ol>
| |
− | </li>
| |
− | <li>
| |
− | <span>The relation between the concentration of benzo[a]pyrene and illumination of RFP</span>
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− | <ol class="article-ol" type="A">
| |
− | <li><span>Method</span></li>
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− | <li><span>Results</span></li>
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− | <li><span>Discussion</span></li>
| |
− | </ol>
| |
− | </li>
| |
− | <ol>
| |
− | </article>
| |
| <article class="article"> | | <article class="article"> |
| <h3 class="article-title">Reference</h3> | | <h3 class="article-title">Reference</h3> |
| <ul class="article-ul"> | | <ul class="article-ul"> |
− | <li>[1] 黄靖芬,李来好,陈胜军,杨贤庆..烟熏食品中苯并(a)芘的产生机理及防止方法smoked foods Mechanism of benzo (a) pyrene and prevention methods</li> | + | <li>[1] Smoked foods Mechanism of benzo (a) pyrene and prevention methods</li> |
− | <li>[2] 降低食品中多環芳香族碳氫化合物含量之作業指引(草案) reduce operating guidelines in food polycyclic aromatic hydrocarbon content of the (draft)</li> | + | <li>[2] Reduce operating guidelines in food polycyclic aromatic hydrocarbon content of the (draft)</li> |
| <li>[3] Basic Information about Benzo(a)pyrene in Drinking Water</li> | | <li>[3] Basic Information about Benzo(a)pyrene in Drinking Water</li> |
| <li>[4] Hadibarata T, Kristanti RA. Identification of metabolites from benzo[a]pyrene oxidation by ligninolytic enzymes of Polyporus sp. S133. J Environ Manage. 2012 11/30;111(0):115-9.(2013 IGEM CUHK)</li> | | <li>[4] Hadibarata T, Kristanti RA. Identification of metabolites from benzo[a]pyrene oxidation by ligninolytic enzymes of Polyporus sp. S133. J Environ Manage. 2012 11/30;111(0):115-9.(2013 IGEM CUHK)</li> |
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| </ul> | | </ul> |
| </article> | | </article> |
| + | </div> |
| + | <div class="link-figure"><img src="https://static.igem.org/mediawiki/2015/0/05/HSNUTAIPEIPROJECT.png"> |
| + | <ul> |
| + | <li><a href="https://2015.igem.org/Team:HSNU-TAIPEI/projectBP">Benzo[A]Pyrene</a></li> |
| + | <li><a href="https://2015.igem.org/Team:HSNU-TAIPEI/projectcopper">Copper</a></li> |
| + | <li><a href="https://2015.igem.org/Team:HSNU-TAIPEI/projectcadmium">Cadmium</a></li> |
| + | <li><a href="https://2015.igem.org/Team:HSNU-TAIPEI/projectmercury">Mercury</a></li> |
| + | <li><a href="https://2015.igem.org/Team:HSNU-TAIPEI/projectlead">Lead</a></li> |
| + | <li><a href="https://2015.igem.org/Team:HSNU-TAIPEI/projectafla">Aflatoxin</a></li> |
| + | </ul> |
| + | </div> |
| + | <div class="side-nav"> |
| + | <h4>Project</h4> |
| + | <ul> |
| + | <li><a href="https://2015.igem.org/Team:HSNU-TAIPEI/Description">Overview</a></li> |
| + | <li><a href="https://2015.igem.org/Team:HSNU-TAIPEI/projectBP">Benzo[A]Pyrene</a></li> |
| + | <li><a href="https://2015.igem.org/Team:HSNU-TAIPEI/projectcopper">Copper</a></li> |
| + | <li><a href="https://2015.igem.org/Team:HSNU-TAIPEI/projectcadmium">Cadmium</a></li> |
| + | <li><a href="https://2015.igem.org/Team:HSNU-TAIPEI/projectmercury">Mercury</a></li> |
| + | <li><a href="https://2015.igem.org/Team:HSNU-TAIPEI/projectlead">Lead</a></li> |
| + | <li><a href="https://2015.igem.org/Team:HSNU-TAIPEI/projectafla">Aflatoxin</a></li> |
| + | </ul> |
| </div> | | </div> |
| | | |
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