Difference between revisions of "Team:East Chapel Hill/Composite Part"
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<li><a href="https://2017.igem.org/Team:East_Chapel_Hill/HP/Silver">HP Silver</a></li> | <li><a href="https://2017.igem.org/Team:East_Chapel_Hill/HP/Silver">HP Silver</a></li> | ||
<li><a href="https://2017.igem.org/Team:East_Chapel_Hill/HP/Gold_Integrated">HP Gold</a></li> | <li><a href="https://2017.igem.org/Team:East_Chapel_Hill/HP/Gold_Integrated">HP Gold</a></li> | ||
| − | <li><a href="https://2017.igem.org/Team:East_Chapel_Hill/Engagement">Engagement</a></li> | + | <li><a href="https://2017.igem.org/Team:East_Chapel_Hill/Engagement">Engagement</a></li> |
| + | <li><a href="https://2017.igem.org/Team:East_Chapel_Hill/Collaborations">Collaborations</a></li> | ||
</ul> | </ul> | ||
</li> | </li> | ||
| − | |||
| − | |||
</ul> | </ul> | ||
</div> | </div> | ||
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<div class="container" style="color:#feffff; margin-top:50px;"> | <div class="container" style="color:#feffff; margin-top:50px;"> | ||
| − | <div class="row | + | <div class="row"> |
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| − | <h1>Best Composite Part</h1> | + | <h1>Best New Composite Part</h1> |
<p style="font-size:18px;"> | <p style="font-size:18px;"> | ||
We have one new composite part: the <b>Fluoride Riboswitch Regulated Chloramphenicol Acetyltransferase Operon. </b> </p> | We have one new composite part: the <b>Fluoride Riboswitch Regulated Chloramphenicol Acetyltransferase Operon. </b> </p> | ||
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| + | |||
| + | <figure> | ||
| + | <img src="https://static.igem.org/mediawiki/2017/e/ef/T--East_Chapel_Hill--design-f5.png" style="width:75%;height:auto;"> | ||
| + | <figcaption>Schematic of the fluoride riboswitch regulated chloramphenicol acetyltransferase operon (CHOP). | ||
| + | |||
| + | </figcaption> | ||
| + | </figure> | ||
| + | |||
| + | <p style="font-size:18px;"> | ||
| + | <br> | ||
| + | Promoter used: <a href="http://parts.igem.org/Part:BBa_J23100"><b><u>BBa_J23100</u></b></a>. | ||
| + | |||
| + | <br> | ||
| + | The restriction enzymes sites make it easy to put in new genes using the Gibson Assembly just order your gene of interest with the correct gibson homology overhangs (15-20nts) and cut with the appropriate restriction enzyme. | ||
| + | <br><br> | ||
| + | |||
| + | HindIII = promoter riboswitch segment<br> | ||
| + | XhoI = new genes<br><br> | ||
| + | |||
| + | This part allows for determining the responsiveness of a fluoride riboswitch and can be used to qualitatively measure fluoride levels. How it works: | ||
| + | |||
| + | <ul style="text-align:left;"> | ||
| + | <li>Using the ΔcrcB <i>E. coli</i> strain, which can accumulate fluoride intracellularly.</li> | ||
| + | <li>The Riboswitch detects fluoride Fluoride activates the chloramphenicol acetyltransferase enzyme, which allows for the growth of bacteria on agar plates with the antibiotic chloramphenicol.</li> | ||
| + | </ul> | ||
| + | </p> | ||
| + | |||
| + | <p style="font-size:18px;"> | ||
| + | This system is amenable for high-throughput screening and can be used to identify fluoride riboswitches with higher affinity to fluoride. However, this system should work with any transcriptional riboswitch. | ||
| + | |||
| + | <br><br> | ||
| + | </p> | ||
| + | |||
| + | |||
| + | </div> | ||
| + | </div> | ||
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</html> | </html> | ||
Latest revision as of 02:52, 2 November 2017
Best New Composite Part
We have one new composite part: the Fluoride Riboswitch Regulated Chloramphenicol Acetyltransferase Operon.
Promoter used: BBa_J23100.
The restriction enzymes sites make it easy to put in new genes using the Gibson Assembly just order your gene of interest with the correct gibson homology overhangs (15-20nts) and cut with the appropriate restriction enzyme.
HindIII = promoter riboswitch segment
XhoI = new genes
This part allows for determining the responsiveness of a fluoride riboswitch and can be used to qualitatively measure fluoride levels. How it works:
- Using the ΔcrcB E. coli strain, which can accumulate fluoride intracellularly.
- The Riboswitch detects fluoride Fluoride activates the chloramphenicol acetyltransferase enzyme, which allows for the growth of bacteria on agar plates with the antibiotic chloramphenicol.
This system is amenable for high-throughput screening and can be used to identify fluoride riboswitches with higher affinity to fluoride. However, this system should work with any transcriptional riboswitch.
