Difference between revisions of "Team:DTU-Denmark/Tour Results"
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<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 the | + | <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 Naja nigricollis. The initial AMC substrate experiment led to a more comprehensive substrate screening experiment that resulted in multiple substrate candidates.</p><br><br> |
| + | |||
| + | <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> <br><br> | ||
| + | |||
| + | <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</h> | ||
| + | <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>. | Read more <a href="https://2017.igem.org/Team:DTU-Denmark/Results">here</a>. | ||
</p><br> | </p><br> | ||
Revision as of 01:44, 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.
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