Results

This experiment has two parts: one tested the effectiveness of chitinase breakdown using our synthesized strain of ChiA and the other tested the ability of said strain tobe mutated and we used mathematical models to predict whether or not enzyme evolution via error prone PCR is possible.

In proving the ability of GST-ChiA-FLAG (our synthesized construct) to secrete chitinase, we used two methods. In the solution assay there was significant chitinolytic activity and the levels of degradation were measured using colorimetric methods and absorbance measurements. (see Figure 1)

The levels of chitinolytic activity were measured using a serial dilution of chitinase to test to see that an increase in chitinase concentration is correlated to increased chitin degradation. As the concentration of chitinase decreases from left to right, there is a decrease in absorbance/color showing that there is, in fact, a direct correlation.

The plate on the right was plated with colonies of bacteria that was transformed with our GST-ChiA-FLAG construct and the plate on the left was plated with bacteria transformed with a negative control (pBAD-D4). There are visible rings and more colonies on the right as there is actual ChiA sequences in those bacteria and the vector used for this construct had ampicillin resistance. The negative control didn’t have a sequence for ChiA but did have ampicillin resistance although there does seem to be a lower survival rate in the negative control colonies regardless.

Caption: this is a banana!

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