Composite

Part:BBa_M50053:Design

Designed by: Amanda Urke, Tofe Alimi, Katherine Vera   Group: Stanford BIOE44 - S11   (2016-12-11)


FRET-based glucose sensor using a glucose binding protein, mRuby3 and cometGFP


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 77


Design Notes

We engineered E. coli to produce a unique protein composed of two fluorophores - mRuby3 and cometGFP - joined by linkers each to the hinge of a glucose binding protein. This construct will undergo a conformational change upon binding to glucose such that the cometGFP portion in an excited state will transfer energy to the acceptor mRuby3 to be emitted. Quantifying this excitation/emission pair reveals the extent to which FRET and thus glucose binding that has occurred.

Source

Key Parts: 

1. mRuby3 Fluorescent Protein - https://parts.igem.org/Part:BBa_M50009 

2. 6 Amino Acid Fusion Protein Linker - https://parts.igem.org/Part:BBa_M50036 

3. Glucose Binding Protein of Thermus thermophilus - https://parts.igem.org/Part:BBa_M50037

References