Reporter

Part:BBa_K1182000:Design

Designed by: Muhammad Hanifi   Group: iGEM13_UI-Indonesia   (2013-09-16)


Split β-galactosidase - ω fragment


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

This is an alpha fragment of beta galactosidase split enzyme. The enzyme beta galactosidase is expressed separately into two fragment, the alpha and omega fragment. When the alpha and omega fragment meets they will exhibit an enzymatic activity similar to the full length expressed beta galactosidase enzyme.

This split reporter can be used as biosensor using ternary complexation mediated protein complementation principle where the interaction of two proteins (in this case the alpha and omega fragment) is mediated by another protein.

This split reporter also can be used as a reporter for protein-fragment complementation assay to test the interaction of two proteins.

Source

The part is fragment of E.coli LacZ gene, acquired by PCR.

References

Stains, C.I., et al. (2010). A General Approach for Receptor and Antibody-Targeted Detection of Native Proteins utilizing Split-Luciferase Reassembly. ACS Chem Biol. 5(10): 943–952. doi:10.1021/cb100143m.

Broome, A.M., et al. (2010). Expanding the utility of Beta-galactosidase complementation: pieceby piece. Mol Pharm. 7(1): 60–74. doi:10.1021/mp900188e.