Difference between revisions of "Team:DTU-Denmark/Tour Results"
| (7 intermediate revisions by 3 users not shown) | |||
| Line 36: | Line 36: | ||
<a href="https://2017.igem.org/Team:DTU-Denmark/Tour_Results" class="tourlink tourlink3">Results</a> | <a href="https://2017.igem.org/Team:DTU-Denmark/Tour_Results" class="tourlink tourlink3">Results</a> | ||
<a href="https://2017.igem.org/Team:DTU-Denmark/Tour_Prototype" class="tourlink tourlink4">Prototype</a> | <a href="https://2017.igem.org/Team:DTU-Denmark/Tour_Prototype" class="tourlink tourlink4">Prototype</a> | ||
| − | <a href="https://2017.igem.org/Team:DTU-Denmark/Tour_Human_Practices" class="tourlink tourlink5"> | + | <a href="https://2017.igem.org/Team:DTU-Denmark/Tour_Human_Practices" class="tourlink tourlink5">H-Practices</a> |
| − | <a href="https://2017.igem.org/Team:DTU-Denmark/Tour_Team_Sponsors" class="tourlink tourlink6">Team | + | <a href="https://2017.igem.org/Team:DTU-Denmark/Tour_Team_Sponsors" class="tourlink tourlink6">Team</a> |
<a href="https://2017.igem.org/Team:DTU-Denmark/Tour_Attributions" class="tourlink tourlink7">Attributions</a> | <a href="https://2017.igem.org/Team:DTU-Denmark/Tour_Attributions" class="tourlink tourlink7">Attributions</a> | ||
</div> | </div> | ||
| Line 52: | Line 52: | ||
<h1 class="bottomborder">Results</h1> | <h1 class="bottomborder">Results</h1> | ||
| − | <p>The AMC experiment showed that we | + | <p>In the pursuit of a possible linker and reporter method for our biosensor, we did a number of experiments. </p> |
| − | We managed to assemble the peptide sequences into the plasmid backbone (pSB1C3). The venom degradation test of the reporter molecules amilCP and β-galactosidase showed no reduction in the colorimetric or enzymatic properties. | + | |
| − | + | <h3>AMC and linker substrate experiments</h3> | |
| − | Instead of the amilCP | + | <p>The AMC (a fluorescent molecule tied with a linker) experiment showed that we were able to detect a significant difference between the Bitis species and <i>Naja nigricollis</i>. The initial AMC substrate experiment led to a more comprehensive substrate screening experiment that resulted in multiple substrate candidates.</p> |
| + | |||
| + | <h3>Assembly</h3> | ||
| + | <p>We managed to assemble the peptide sequences into the plasmid backbone (pSB1C3), producing a number of basic parts. Overall, we submitted 11 linker parts and 2 composite parts to the biobrick registry.</p> | ||
| + | |||
| + | <h3>Stability in presence of venom</h3> | ||
| + | <p>The venom degradation test of the reporter molecules amilCP and β-galactosidase showed no reduction in the colorimetric or enzymatic properties.</p> | ||
| + | |||
| + | <h3>Improving a previous part</h3> | ||
| + | <p>Furthermore, we improved the part BBa_K592009 (purple-blue chromoprotein amilCP from Uppsala, Sweden) by adding a His-tag on its C-terminus. The expression of color was not reduced and the color protein could easily be retained by a His-tag purification. </p> | ||
| + | |||
| + | <h3>Further improvement</h3> | ||
| + | <p>To further improve our diagnostic device, a reduction on the response time for the result was undertaken. Instead of the amilCP reporter, the enzyme β-galactosidase was to be attached to the substrate linker. However, there was no successful assembly of ScAvidin with the linker to β-galactosidase. | ||
| + | <br><br> | ||
| + | Read more <a href="https://2017.igem.org/Team:DTU-Denmark/Results">here</a>. | ||
| + | </p><br> | ||
| Line 67: | Line 82: | ||
<ul> | <ul> | ||
<li class="rightnavbarbtn"> | <li class="rightnavbarbtn"> | ||
| − | <a href="#results"> | + | <a href="#results"></a> |
</li> | </li> | ||
</ul> | </ul> | ||
Latest revision as of 02:20, 2 November 2017
Results
In the pursuit of a possible linker and reporter method for our biosensor, we did a number of experiments.
AMC and linker substrate experiments
The AMC (a fluorescent molecule tied with a linker) experiment showed that we were able to detect a significant difference between the Bitis species and Naja nigricollis. The initial AMC substrate experiment led to a more comprehensive substrate screening experiment that resulted in multiple substrate candidates.
Assembly
We managed to assemble the peptide sequences into the plasmid backbone (pSB1C3), producing a number of basic parts. Overall, we submitted 11 linker parts and 2 composite parts to the biobrick registry.
Stability in presence of venom
The venom degradation test of the reporter molecules amilCP and β-galactosidase showed no reduction in the colorimetric or enzymatic properties.
Improving a previous part
Furthermore, we improved the part BBa_K592009 (purple-blue chromoprotein amilCP from Uppsala, Sweden) by adding a His-tag on its C-terminus. The expression of color was not reduced and the color protein could easily be retained by a His-tag purification.
Further improvement
To further improve our diagnostic device, a reduction on the response time for the result was undertaken. Instead of the amilCP reporter, the enzyme β-galactosidase was to be attached to the substrate linker. However, there was no successful assembly of ScAvidin with the linker to β-galactosidase.
Read more here.
Previous: Approach Next: Prototype
