Generator

Part:BBa_K1316014:Design

Designed by: Joan Cortada Garcia   Group: iGEM14_TU_Delft-Leiden   (2014-09-29)


pRhamnose regulating csgB + J23110 constitutive Anderson promoter regulating csgA with a His tag


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 516
    Illegal NheI site found at 551
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Many more CsgA protein subunits are required compared to CsgB in curli formation.

A His tag was added into the CsgA protein for the mentioned purposes (The His tag present in the CsgA protein has two functions: on one hand it allows easy protein purification of the CsgA protein; on the other hand, it enables the formation of conductive curli by adding gold nanoparticles (AuNP) with Nickel-Nitrilotriacetic-acid (NTA-Ni) attached to them. This last part containing Nickel would bind to the Histidines of the curli fibril, whereas the gold attached to it will make the environment more conductive.)


Source

csgA and csgB were PCRed out of the K540000 Lyon 2011 iGEM part.

References