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.