Difference between revisions of "Part:BBa K1993020"

Latest revision as of 19:23, 17 October 2016

dTomato

The dTomato protein is a fluorescent protein dimer, created by direct evolution of the wild-type DsRed, from Discosoma sp. [1] The dTomato protein is a fluorescent dimer protein that emits orange-red light when it is excited by green-yellow light. It is preferable to use especially in self-made fluorometry tests because the excitation wavelengths and the emission wavelengths don't overlap as much as in other fluorescent proteins. The dTomato excitation peak is at 554 nm and 50% of it is at 510 nm. Also, its emission peak is at 581 nm and its 50% emission at 629 nm.

In our project, in order to ensure our engineered MSCs were locating in inflamed lesions, we introduced dTomato gene. Firstly, we acquired it from a synthetic constructed plasmid we purchased from Addgene. (Figure 1) Secondly, we combined dTomato with other genes by Gateway technology and constructed a composite part, Luciferase-T2A-dTomato-T2A-hFTH BBa_K1993006. Thirdly, we transduced this composite part into MSCs. Finally, we confirmed whether dTomato were expressed in vitro. (293FT cells and MSCs) (Figure 2, 3)

Figure 1 DNA of dTomato was shown in agarose gel electrophoresis.

Figure 2 dTomato was expressed in 293FT cells.

Figure 3 dTomato was expressed in MSCs.

References

[1] Shaner et al, Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein, 2004 Sequence and Features