Part:BBa_M50084:Experience

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Summary: Measuring the responsiveness of the serotonin receptor using dynamic range experiment with varying serotonin concentrations.

Documentation: We tested whether the transformed Saccharomyces cerevisiae cells successfully expressed the serotonin receptor by measuring fluorescence after they were grown in SC media containing 2% galactose and varying concentrations of serotonin. We used a strain of S. cerevisiae that had a pAga1 promoter tied to an EGFP reporter. We used an initial S. cerevisiae cell density of OD600 of 0.1 and them in 2% galactose and SC media to induce construct expression. We then allowed our cells to incubate for 2 days at 30°C at 750 rpm, where they grew to an OD600 of around 0.2. We then added varying concentrations of serotonin, ranging from 0.1 µM to 1000 µM (specifically, we tested at 0.1, 1.0, 10.0, 100.0, and 1000.0 µM). We also included a negative control that did not receive any serotonin, and a positive control that had 1 uM alpha factor instead of serotonin (see part BBa_K1829005).

Results:

Conclusions: We normalized the GFP measurement with the OD600 of each sample and observe a slight increase in normalized fluorescence as the concentration of serotonin increase. All of our experimental samples show greater normalized fluorescence readings than the negative control (the leftmost column). While the negative control has a greater normalized fluorescence than the positive control, the variance of each sample is such that there is no significant difference between any two samples. Our results seem to indicate that, at the very least, the presence of serotonin caused some fluorescence in the pSery strain, though the high variance of the shown samples makes it difficult to gain great insight into a general trend.

Stanford location:

Plasmid name: pSery DNA2.0 Gene #: 309066 Organism: Saccharomyces cerevisiae Device Type: Sensor Barcode numbers: 0133009654, 0133011784 Box Label: BIOE44 F17

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