Part:BBa_K4375007

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/