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Part Characterization

BBa_K567018 - PT7-GFP-TAG-RFP

This part was characterized by Team ULaval in the cell-free expression system myTX-TL from ArborBioscience. The fluorescence intensity of this part had only been characterized in vivo, and no data was available for the use of this part outside of a cellular system. By expressing this part in a cell-free commercial media, and measuring the intensity of GFP fluorescence, we hope to provide other teams with data that shows that this part can be used outside of cellular systems. As shown by figure 3, fluorescence was observed after 75 minutes, attained significant levels after about 5h and peaked after 16h.

While this part was not designed with cell-free systems in mind, our data shows that it is indeed appropriate for use. Using this part in cell-free systems could ease the purification of this part if large quantities of the purified protein are necessary for a given experiment.

**Figure 1.** Fluorescence curve for BBa_K567018. Fluorescence intensity was measured and corrected according to the IGEM data analysis table v2. However, as these fluorescence measurements were carried outside of cells, no OD600 was calculated. Fluorescence is therefore only corrected using the fluorescein reference curve.

BBa_I746907 - T7 promoter driving 6-his tagged P7 GFP

This part was characterized by Team ULaval in the cell-free expression system myTX-TL from ArborBioscience. This part had never been characterized, and no fluorescence or expression data were available. By expressing this part in a cell-free commercial media, and measuring the intensity of GFP fluorescence, we hope to demonstrate that this part is indeed functional, and usable inside a cell-free expression system. As shown by figure 1, fluorescence was observed after 75 minutes, attained significant levels after about 5h and peaked after 16h.

Our data shows that this part is compatible with the cell-free expression system used. Using this part in cell-free systems could ease the purification of this part if large quantities of the purified protein are necessary for a given experiment.

**Figure 2.** Fluorescence curve for BBa_I746907. Fluorescence intensity was measured and corrected according to the IGEM data analysis table v2. However, as these fluorescence measurements were carried outside of cells, no OD600 was calculated. Fluorescence is therefore only corrected using the fluorescein reference curve.

Team:ULaval/Contribution