Part:BBa_K2083010

Coding region for GFP reporter. The start codon has been mutated to ACG to decrease, and hopefully prevent transcription. Overall, in experiments there was decreased production of GFP compared to BBa_K2083009.

Figure 1: Wild Type ATG GFP Reporter Fluorescence

Figure 2: Mutated Start Codon GFP Reporter Fluorescence

The data indicates relatively high efficiency in the ATG/ACG reporters. According to the microscopy data the ATG reporter showed 70.57% of fluorescence compared to the positive control, while the ACG reporter only gave 4.21%. Therefore, the mutated start codon is proven to be able to produce the difference between the two versions of the reporter.

Figure 3: Efficiency of ATG/ACG reporters compared to positive control. Error bars represent standard error between normalized results from 3 biological replicates of the experiment.

Figure 4: ATG/ACG reporters efficiency confirmation by flow cytometry. The difference between the wild type and mutant reporter is consistent with the microscopy data.

For how the values were calculated see this: https://static.igem.org/mediawiki/2016/c/c7/T--WPI_Worcester--Imaging_Analysis.pdf

For the fluorescence values see here:

https://static.igem.org/mediawiki/2016/d/d2/T--WPI_Worcester--ATG_Trial1.pdf: Trial 1

https://static.igem.org/mediawiki/2016/1/1a/T--WPI_Worcester--ATG_Trial2.pdf: Trial 2

https://static.igem.org/mediawiki/2016/8/81/T--WPI_Worcester--ATG_Trial3.pdf: Trial 3