Composite

Part:BBa_K2476001:Design

Designed by: Vivian Hu   Group: iGEM17_Pittsburgh   (2017-10-27)


WT CheY + Dronpa fusion protein regulated by IPTG


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1302
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This is a composite part that includes the WT CheY + Dronpa protein coding region encoded in part BBa_K2476012 and the Dronpa protein coding region encoded in part BBa_K2476010.

Uses an IPTG inducible promoter with RBS (BBa_J04500), WT CheY + Linker/Dronpa/Linker at 60-61 (BBa_K2476012), Dronpa (BBa_K2476010), and a double terminator (BBa_B0015). Will express WT CheY + Dronpa fusion protein when induced with IPTG.CheY is a member of this family that interacts with FliM found on the flagellar motor. This interaction directly regulates the swimming patterns of Ecoli. Another member of the pathway, CheY A, controls the activation of CheY by transferring a phosphate group. The active form of CheY-P interacts with FliM causing a tumble. When CheY is inactive, the protein does not interact with FliM and the bacteria runs. Therefore, in the presence of a repellant, CheA phosphorylates CheY and the cell tumbles. In the presence of an attractant, CheA does not transfer a phosphate group to CheY and the cell swims.

Source

IPTG inducible promoter with RBS (BBa_J04500), WT CheY + Linker/Dronpa/Linker at 60-61 (BBa_K2476012), Dronpa (BBa_K2476010), and a double terminator (BBa_B0015)

References