Part:BBa_K1993006
Luciferase-T2A-dtomato-T2A-hFTH
Firefly (Photinus pyralis) Luciferase (Details can be seen from BBa_K1993018) is a kind of oxidative enzyme that produce bioluminescence, which is distinct from a photoprotein. It is able to oxidize luciferin and produce detectable bioluminescence. It’s convenient to observe biological processes, especially allowing for non-invasive observation of cells.
However, the shortage of such technique is also obvious: cost of substrate luciferin is relative expensive and the intensity of measured signal by in vivo imaging may be affected by various factors which weaken its accuracy. Furthermore, it is not always necessary to observe cells in vivo since in biological researches, in vitro observation is more frequent.
Previously, iGEM07_Ljubljana had already designed the part BBa_I712019, containing the coding sequence of Firefly Luciferase which had fulfilled function of observation in vivo.
In order to improve its function, we constructed a new device enabling us to observe biological processes both in vivo and in vitro. We expanded the previous part by adding two more genes: dTomato (Details can be seen from BBa_K1993020) used for observation in vitro conveniently and easily; hFTH (See details in BBa_K1993021), another protein that could be observed in vivo by MRI. (Figure 1) What’s more, in order to make sure the expression of all three proteins, we added a T2A part (Details can be seen from BBa_K1993019) between every two protein coding sequences. (Figure 1)
Figure 1 Constitution of our new device.
Since iGEM had helped us confirm the function of Firefly Luciferase, we only tested the expression of dTomato. See results below (Figure 2, Figure3)
Figure 2 Expression of dTomato in 293FT Cells.
Figure 3 Expression of dTomato in MSCs.
As for hFTH, we would test it in non-human primates first, then we may apply our device for MSCs preclinical researches in the future.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 808
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