Team:SCSU-New Haven/Experiments
Experiments
You have the parts. You do the tests to get the numbers.
Detector Assay
The Detector assay will be done by placing GFP downstream of Psal and exposing the cells to 3-Methylsalicylate. The fluorescence response will be measured. We would like to see a response from 0.1uM and less of 3-Methylsalicylate as this was the amount required to produce a response in the paper we obtained the sequence from.
Feedback Assay
The Assay for the Feedback part will be similar to the Detector Assay. The Feedback part will be place downstream of Psal and upstream of GFP and fluorescence will be measured. If the part is working as intended, we would see a rise in fluorescence to a steady state. This means the feedback loop is turning the Psal promoter on and keeping it on as intended.
Color Reporter Assays
The Assay for color reporter will be as the above assays substituting the GFP for the Color Reporter part. The production of Indole will be measured using a spectrophotometer. We should expect to see production of Indole comparable to fluorescence.
Degrader Assay
The Degrader would then be added and measurements of Indigo Dye would be taken. Comparing these results to the production of indole would tell us how well the Degrader enzymes convert indole to indigo dye in the system. We could also assay the conversion of 1-MethylNapthalene to 3-Methylsalicylate by comparing the fluorescence produced when 1-MethylNapthalene is added compared to 3-Methylsalicylate.
