title: Cell surface display of poliovirus receptor on Escherichia coli, a novel method for concentrating viral particles in water.
abstract: The lack of efficient methods for concentrating viruses in water samples leads to underreporting of viral contamination in source water. A novel strategy for viral concentration was developed using the expression of target virus receptors on bacterial cells. Poliovirus type 1, the most studied enterovirus, was used as a surrogate for enteric viruses. The human poliovirus receptor (hPVR) gene was expressed on the surface of Escherichia coli cells by using the ice nucleation protein (INP) gene. The hPVR gene was ligated to the 3' end of the INP gene after the removal of the stop codon. The resulting open reading frame (ORF) was used for the projection of hPVR onto the outer membrane of E. coli. Gene expression was tested by SDS-PAGE, Western blot, and dot blot analyses, and virion capture ability was confirmed by transmission electron microscopy. The application of engineered E. coli cells for capturing viruses in 1-liter samples of source and drinking water resulted in 75 to 99% procedural recovery efficiency. Cell surface display of viral receptors on bacterial cells opens a new prospect for an efficient and inexpensive alternative tool for capturing and concentrating waterborne viruses in water samples.
authors.affiliation: Department of Civil, Environmental and Sustainable Engineering, Arizona State University, Tempe, AZ 85287, USA. abbaszadegan@asu.edu
authors.forename: Morteza
authors.initials: M
authors.lastname: Abbaszadegan
authors.forename: Absar
authors.initials: A
authors.lastname: Alum
authors.forename: Hamed
authors.initials: H
authors.lastname: Abbaszadegan
authors.forename: Valerie
authors.initials: V
authors.lastname: Stout
mesh_terms: D000954:Antigens, Surface / Q000096:biosynthesis* / Q000235:genetics; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis / Q000235:genetics; D015153:Blotting, Western; D002462:Cell Membrane / Q000737:chemistry* / Q000821:virology; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000737:chemistry* / Q000235:genetics; D015870:Gene Expression; D015151:Immunoblotting; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D017955:Poliovirus / Q000235:genetics / Q000302:isolation & purification*; D011991:Receptors, Virus / Q000032:analysis* / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics; D014771:Virion / Q000737:chemistry* / Q000648:ultrastructure; D014867:Water; D014871:Water Microbiology; D014876:Water Pollution
chemical_list: D000954:Antigens, Surface; D001425:Bacterial Outer Membrane Proteins; D008565:Membrane Proteins; D011991:Receptors, Virus; D011994:Recombinant Proteins; C063113:ice nucleation protein; C066100:poliovirus receptor; D014867:Water