Coding

Part:BBa_K4164004:Design

Designed by: Bo Cheng   Group: iGEM22_NAU-CHINA   (2022-09-30)


dimerization-dependent red fluorescent protein-A1(ddRFP-A1)


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]


Design Notes

ddRFP-A1 is an RFP variant with weak fluorescence derived from the directed evolution of dTomato gene. It has a chromogenic group,and can exhibit weak fluorescence in the monomeric state.

ddRFPA1 can form a heterodimer with ddRFPB1, increasing the fluorescence intensity ten times more than the dissociation state. The utility of ddRFP-A1B1 has been demonstrated in three applications, including the detection of a protein-protein interaction in vitro, imaging of the reversible Ca2+-dependent association of calmodulin, and imaging of caspase-3 activity during apoptosis.

In our project, we fused ddRFP-A1 and ddRFP-B1 to the C-terminus of FXR and RXR, respectively. Taking advantage of the characteristics that RXR can bind to the FXR as a heterodimer, using FXR as the receptor, and when the downstream ddRFP-A1 and ddRFP-B1 are correspondingly driven to dimerize and exhibit red fluorescence.

Source

The gene sequence of this part was obtained from NCBI, and its GenBank Accession number is AFH02976.1

References