Part:BBa_K1206000:Experience

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Applications of BBa_K1206000

Our project submission BBa_6120600, is designed with a glycine serine amino acid link between the degradation enzyme qsda-1 and GFP reporter. Because of this, when the enzyme begins degrading the autoinducer AHL in the surrounding environment, our plasmid will fluoresce green. For this assay, we used a Tecan plate reader to measure relative fluorescence given off by our modified E. Coli cell when AHL is prominent in the cell’s environment. To be sure we are getting correct results, we decided to measure the relative fluorescence given off by our project cell when it is uninduced, our project cell when it is induced by the autoinducer AHL, and cells of the green fluorescence protein that will be constitutively be expressing fluorescence. The hypothesized results before the assay was preformed were that the GFP would show the most relative fluorescence. That the uninduced project cells will show little to no fluorescence, and the induced project cells will have a relative fluorescence in between the two.

Before we could run the assay, we had to look up the emission wavelength and the excitation wavelength from the iGEM parts registry data base for the specific green fluorescence protein we chose from the iGEM biobrick. These values were 395 nm for emission and 515 nm for excitation wavelength. These values were submitted into the Tecan plate reader computer program for accurate readings. When we ran our plate reader, the average fluorescence for each were determined, and the results fit in with our hypothesis. The GFP cells fluoresced the most, the uniduced cells fluoresced the least, and the induced project cells relative fluorescence was in the midde. The average relative fluorescence of the three are as follows and are measured as a numerical value of relative fluorescence given off.

Green Flourescence proteins : 42,205 Induced E. Coli cell : 31,248 Uninduced E. Coli cell: 24,025

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