Difference between revisions of "Part:BBa K1993020"

 
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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.
 
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)
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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)
  
 
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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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]