Difference between revisions of "Part:BBa K4375007"
Line 9: Line 9:
 
==Usage and Biology==
 
==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.
+
The INP (Ice Nucleation Protein) is a membrane-bound protein from bacteria, like Pseudomonas and Erwinia, and can be used as an anchor motif for cell surface display. INPNC contains the N -and C-terminal domains of this protein in a truncated form. Through codon optimization, we made it compatible with E.Coli to be expressed.  
  
  
Line 19: Line 19:
  
  
−
==References==
+
==Reference==
−
 
+
−
https://pubmed.ncbi.nlm.nih.gov/16817238/
+
  
 +
Wu PH, Giridhar R, Wu WT. Surface display of transglucosidase on Escherichia coli by using the ice nucleation protein of Xanthomonas campestris and its application in glucosylation of hydroquinone. Biotechnology and bioengineering. 2006 Dec 20;95(6):1138-47.
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Revision as of 10:24, 10 October 2022


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 (Ice Nucleation Protein) is a membrane-bound protein from bacteria, like Pseudomonas and Erwinia, and can be used as an anchor motif for cell surface display. INPNC contains the N -and C-terminal domains of this protein in a truncated form. Through codon optimization, we made it compatible with E.Coli to be expressed.



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]


Reference

Wu PH, Giridhar R, Wu WT. Surface display of transglucosidase on Escherichia coli by using the ice nucleation protein of Xanthomonas campestris and its application in glucosylation of hydroquinone. Biotechnology and bioengineering. 2006 Dec 20;95(6):1138-47.