Coding

Part:BBa_K4164998

Designed by: Bo Cheng   Group: iGEM22_NAU-CHINA   (2022-10-01)
Revision as of 14:03, 12 October 2022 by BoCheng (Talk | contribs)


ddRFPA1-ddRFPB1

The red fluorescent protein heterodimer is formed by the polymerization of two monomeric proteins ddRFPA1 and ddRFPB1.The monomer is derived from the directed evolution of dTomato gene. ddRFPA1 exhibits weak fluorescence, ddRFPB1 has no fluorescence.ddRFP-A1 can form a heterodimer with ddRFP-B1, 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.

We connected ddRFPA1 and ddRFPB1 by a flexible linker(3*GGGGS) to verify the feasicility of ddRFPA1 and ddRFPB1. Following overnight incubation at 37℃, we can see the bright red fluorescence (Fig.1). In this case, we chose ddRFPA1 and ddRFPB1 as our report device.


Fig.1 Fluorescence image of E. coli expressing ddRFPA1-ddRFPB1 and control.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1174
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 733
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None