Coding

Part:BBa_K4375007

Designed by: Zska Csorba   Group: iGEM22_ELTE   (2022-09-28)
Revision as of 09:37, 7 October 2022 by Golaritsagnes (Talk | contribs)


Codon optimalised INPNC for Surface Display

INPNC codes for N- and C- terminal domain of Ice Nucleation Protein (INP) from Pseudomonas syringae, and it can be used for displaying proteins on bacteria's outer membrane.


Usage and Biology

The INP, a membrane-bound protein from ice-nucleation active bacteria, like Pseudomonas, Xanthomonas, Erwinia, is capable of catalyzing the formation of ice in supercooled water. INP normally resides on the surface of cells via the glycosylphosphatidylinositol (GPI)-anchor, this is quite unique for prokaryotes since this motif is normally found only in eukaryotic cells. INP is composed of three distinct structural domains, namely the specific N-terminal region (INPN) that seems to interact with the phospholipids moiety of the outer membrane, the C-terminal domain (INPC) that is highly hydrophilic and exposed to the outermost membrane, and the central domain that is composed of repeating an 8-, 16-, and 48-residue which acts as the template for ice nucleation. This protein was expressed in E. coli, more than 90% of INP was found in the outer membrane, which means that INP is compatible with the protein secretion machinery of host cells.



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 72
    Illegal NgoMIV site found at 405
    Illegal AgeI site found at 823
  • 1000
    COMPATIBLE WITH RFC[1000]


References

https://pubmed.ncbi.nlm.nih.gov/16817238/


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