Coding

Part:BBa_K2833008

Designed by: Wanji Li   Group: iGEM18_BJRS_China   (2018-10-04)


J23104+B0032+INP+YFP

Ice nucleation protein(INP)is a secretory outer membrane protein from Pseudomomas syringae P. flurorescens and several other Gram—negative bacteria.In the bacterial surface display technology，INP is widely used as a functional carrier protein.

Design

We got the INP from BBa_K523013 constructed by iGEM11_Edinburgh. To test the surface display efficiency of INP, we added YFP to the downstream of INP, and expressed the fusion protein under the control of J23104 and B0032.

Microscopy result

We observed the fluorescence under the super-resolution microscope. As figure 1 shows, The intracellular expression of sfGFP displayed the rode-shape of E.coli, while the signal of surface displayed YFP showed the dotted pattern, which suggested that the YFP were gathered and distributed at the surface of E.coli..

Fig.1 super-resolution microscopy of INP-YFP Left: intracellular expression of GFP; Right: surface display of YFP via INP


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 467
  • 1000
    COMPATIBLE WITH RFC[1000]


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