Part:BBa_K1497007:Experience

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Applications of BBa_K1497007

The iGEM Team TU Darmstadt 2014 created the naringenin biosynthesis operons under the control of the T7 promoter BBa_I712074 and the strong constitutive promoter BBa_J23100, respectively . They measured the naringenin production after a 16 h incubation time with the naringenin biosensor BBa_K1497020. The cell pellets from E. coli BL21(DE3) – pSB1C3-fdeR-gfp with and without T7-naringenin operon (BBa_K1497017) are shown in figure 3. Only in the cell pellet with BBa_K1497017 exhibited GFP fluorescence. The Darmstadt team was also able to measure the GFP fluorescence quantitatively and to calculate with the help of a calibration curve for the naringenin sensor the production yield of both operons (Figure 4). For BBa_K1497017 was 3 µM naringenin calculated and for the operon with the constitutive promoter BBa_J23100 (BBa_K1497016) was 1.9 µM naringenin calculated.

iGEM TU Darmstadt 2014 :)

Figure 2 Cell pellets with and without T7-Naringenin operon from E. coli BL21(DE3)-pSB1C3-fdeR-gfp. By using ultraviolet light the pellet containing the naringenin operon shows a GFP fluorescence.

Figure 1 Fluorescence of cells with and without the T7-naringenin operon BBa_K1497017 from E. coli BL21(DE3)-pSB1C3-fdeR-gfp and J23100-naringenin operon (BBa_K1497016) from E. coli Top10-pSB1C3-fdeR-gfp, respectively. E. coli BL21(DE3)-pSB1C3-fdeR-gfp without T7-naringenin operon showed no detectable fluorescence. Only in the cells with the functional operon is the GFP fluorescence measurable. The estimated yields are 3 µM for BBa_K1497017 and 1,9 µM for BBa_K1497016.

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