Part:BBa_K1055001:Experience

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Applications of BBa_K1055001

Figure 1. Overlay of exitation spectrum (dashed line) and emission (solid line) of mKate and LSSmOrange

LSSmOrange is an orange fluorescent protein that we want to use for FRET, we us it as a donor. LSSmOrange has an excitation maximum by 437 nm and an emission maximum by 572 nm. The excitation maximum is at 415 nm, another at 453 nm and our measured emission maximum is at 564 nm (Fig. 2). This aberration changes nothing on the FRET system. The excitation maximum is big enough to stimulate mKate, the other fluorescent protein (Fig. 1). Of course, we also checked the emission and excitation of our BL21(DE3) cells (data not shown) and we can eliminate the theories that these cells disturb this florescence measurement.

Figure 2. Excitation spectrum (dashed line) and emission spectrum (solid line) of LSSmOrange with marked maximums

As shown in Fig. 4, the expression of LSSmOrange is not toxic towards the bacterial host, E. coli BL21(DE3).

Figure 3. Cells grown in GFP-medium containing the arabinose promoter regulating GFP expression. Induced at time=0h.

Figure 4. Cells grown in GFP-medium containing the arabinose promoter regulating GFP expression. Induced at time=0h.

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