Difference between revisions of "Part:BBa K1993020"
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− | 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 [https://parts.igem.org/Part:BBa_K1993006 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) | ||
+ | |||
+ | <html> | ||
+ | <img src="https://static.igem.org/mediawiki/2016/d/d2/T--SYSU-MEDICINE--dTomato-1.jpeg" style="width:200px" ></a> | ||
+ | |||
+ | </html> | ||
+ | |||
+ | '''Figure 1 DNA of dTomato was shown in agarose gel electrophoresis.''' | ||
+ | |||
+ | |||
+ | <html> | ||
+ | <img src="https://static.igem.org/mediawiki/2016/4/45/T--SYSU-MEDICINE--dTomato-2.png" style="width:200px" ></a> | ||
+ | |||
+ | </html> | ||
+ | |||
+ | '''Figure 2 dTomato was expressed in 293FT cells.''' | ||
+ | |||
+ | |||
+ | <html> | ||
+ | <img src="https://static.igem.org/mediawiki/2016/b/be/T--SYSU-MEDICINE--dTomato-3.png" style="width:200px" ></a> | ||
+ | |||
+ | </html> | ||
+ | |||
+ | '''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 | ||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
===Usage and Biology=== | ===Usage and Biology=== |
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
- 10COMPATIBLE WITH RFC[10]
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- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]