title: Functional display of foreign protein on surface of Escherichia coli using N-terminal domain of ice nucleation protein.
abstract: We investigated the ability of the N-terminal domain of InaK, an ice nucleation protein from Pseudomonas syringae KCTC1832, to act as an anchoring motif for the display of foreign proteins on the Escherichia coli cell surface. Total expression level and surface display efficiency of green fluorescent protein (GFP) was compared following their fusion with either the N-terminal domain of InaK (InaK-N), or with the known truncated InaK containing both N- and C-terminal domains (InaK-NC). We report that the InaK-N/GFP fusion protein showed a similar cell surface display efficiency ( approximately 50%) as InaK-NC/GFP, demonstrating that the InaK N-terminal region alone can direct translocation of foreign proteins to the cell surface and can be employed as a potential cell surface display motif. Moreover, InaK-N/GFP showed the highest levels of total expression and surface display based on unit cell density. InaK-N was also successful in directing cell surface display of organophosphorus hydrolase (OPH), confirming its ability to act as a display motif.
authors.affiliation: Division of Molecular and Life Sciences, Pohang University of Science and Technology, Pohang 790-784, Korea.
authors.forename: Lin
authors.initials: L
authors.lastname: Li
authors.forename: Dong Gyun
authors.initials: DG
authors.lastname: Kang
authors.forename: Hyung Joon
authors.initials: HJ
authors.lastname: Cha
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D044224:Pseudomonas syringae / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry*; D014162:Transfection / Q000379:methods*
chemical_list: D001425:Bacterial Outer Membrane Proteins; D011993:Recombinant Fusion Proteins; C063113:ice nucleation protein