Part:BBa_K2552003

Ice Nucleation Protein (INP)

Full-length Ice Nucleation Protein (INP) is a kind of cell-surface display system using N-terminal domain.

Usage and Biology

Full-length ice nucleation protein (INP) is an outer membrane protein found in several Gram-negative bacteria that include Pseudomonas syringae, Erwinia herbicola, Xanthomonas campestris and Pseudomonas fluorescens.

All identified full-length INPs are comprised of three distinct structural domains distinguished as the N-terminal domain (15%), the C-terminal domain (4%) and the central repeating domain (81%). The full-length INP N-terminal domain, which contains three or four transmembrane spans and responsible for targeting to the cell surface, can be used to display foreign proteins on the Escherichia coli cell surface. The C-terminal domain is a relatively hydrophilic terminus comprised of 49 amino acids. The central repeating domain (CRD) is composed of contiguous repeats of 8, 16, or 48 residues, which acts as a template for ice crystal formation . As the full-length INP is quite large (1200 to 1500 amino acids) and neither the C-terminal domain nor the CRD of the full-length INPs harbours signal peptide sequences, therefore, the InpN (the N-terminal domain) or the InpNC (containing only the N- and C-terminal portion) was usually used as the anchoring motif to direct translocation of foreign proteins to the cell surface or the cell periplasm.

In our team, we use the N-terminal domain of full-length INP (called INP in our project) as an anchoring motif to display antibody and only N-terminal sequence is submitted in this part.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 535
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 417
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 238