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>
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           <li><a href="https://2017.igem.org/Team:East_Chapel_Hill/Engagement">Engagement</a></li>       
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          <li><a href="https://2017.igem.org/Team:East_Chapel_Hill/Collaborations">Collaborations</a></li>
 
         </ul>
 
         </ul>
 
         </li>
 
         </li>
        <li><a href="https://2017.igem.org/Team:East_Chapel_Hill/Collaborations">Collaborations</a></li>
 
 
 
     </ul>
 
     </ul>
 
   </div>
 
   </div>

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.

Schematic of the fluoride riboswitch regulated chloramphenicol acetyltransferase operon (CHOP).


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.