Team:UFlorida/Results

Results

Most of our time on the project was spent trying to get all three genes into one cell. We succeeded in transforming cells with three plasmids, theoretically containing ARO9, ARO 10 and ADH1, but were unable to confirm the presence of ARO9.

We transformed plasmids containing ARO10 in pETDUET-1 and ADH1 in pCOLADUET-1 in BL21 cells, but were unable to confirm the presence of ARO9 in a DUET vector either through gel electrophoresis or through Sanger Sequencing. ARO9 encodes aromatic animo acid transferase II, which is the first enzyme in the tryptophol pathway. ARO10 in pETDUET-1 and ADH in pCOLADUET-1 were validated using Sanger Sequencing by Genewiz®

We ran an HPLC Standard of tryptophol in King's Medium B. Here, tryptophol shows an elution peak at 162 m/z. We are planning to conduct HPLC using a filtered bacterial culture, and compare the graph generated to the graph of our tryptophol standard. The HPLC results should be available for our poster and presentation. Currently, we are conducting Kirby Bower tests on a fungus in the Chytridiomycota family using tryptophol for one plate, water for one plate, and liquid cultures of our bacteria for another plate. We expect to have the results from these tests for our poster and presentation.