Difference between revisions of "Part:BBa K1688010"
 
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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 (Figure 1). Characterization has been made with the Anderson promoters BBa_J23101(this construct) and BBa_J23110 (construct BBa_K1688011)
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__NOTOC__
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<partinfo>BBa_K1688010 short</partinfo>
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Assembly of Anderson promoter BBa_J23101 and RBS + dTomato (BBa_K1688004). dTomato has not been bricked previously.
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<partinfo>BBa_K1688010 SequenceAndFeatures</partinfo>
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[[File:uppsala_dtomato.jpg]]
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Figure 1. dTomato expressed in ''E. coli'' DH5-alpha with an
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inducible promoter.
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== Usage and Biology==
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The dTomato protein is a fluorescent protein dimer, created by direct evolution of the wild-type DsRed, from Discosoma sp. (Shaner et al, - Improved monomeric red, orange and yellow fluorescent proteins derived from ''Discosoma sp.'' red fluorescent protein, 2004). 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 (Figure 2).
  
 
                           [[File:Uppsala2015_dTomato_Chromatogram.png]]
 
                           [[File:Uppsala2015_dTomato_Chromatogram.png]]
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Figure 1. Excitation (blue curve) and emission (red curve) spectra for the dTomato fluorescent protein (the graph is designed with the following tool: https://www.chroma.com/spectra-viewer)
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Figure 2. Excitation (blue curve) and emission (red curve) spectra for the dTomato fluorescent protein (the graph is designed with the following tool: https://www.chroma.com/spectra-viewer)
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== References ==
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Nathan C Shaner, Robert E Campbell, Paul A Steinbach, Ben N G Giepmans, Amy E Palmer & Roger Y Tsien. “Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein”, 2004

Latest revision as of 18:07, 18 September 2015

dTomato (inc RBS, J23101 promoter)

Assembly of Anderson promoter BBa_J23101 and RBS + dTomato (BBa_K1688004). dTomato has not been bricked previously.


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Uppsala dtomato.jpg

Figure 1. dTomato expressed in E. coli DH5-alpha with an inducible promoter.


Usage and Biology

The dTomato protein is a fluorescent protein dimer, created by direct evolution of the wild-type DsRed, from Discosoma sp. (Shaner et al, - Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein, 2004). 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 (Figure 2). 

                         Uppsala2015 dTomato Chromatogram.png

Figure 2. Excitation (blue curve) and emission (red curve) spectra for the dTomato fluorescent protein (the graph is designed with the following tool: https://www.chroma.com/spectra-viewer)

References

Nathan C Shaner, Robert E Campbell, Paul A Steinbach, Ben N G Giepmans, Amy E Palmer & Roger Y Tsien. “Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein”, 2004