title: Comparative imaging of a bacterial surface-located GFP fusion protein by epifluorescence and scanning near-field optical microscopy.
abstract: IcsA is an autotransporter protein that plays a role in the virulence of Shigella bacteria. We have examined the cellular localization of a fusion of an IcsA fragment to the green fluorescent protein (GFP) expressed in Escherichia coli using a dual epifluorescence and scanning near-field optical microscope. By combining the data obtained from far-field with near-field microscopy of the same sample, discrimination between surface-bound fusion proteins and fusion proteins located in the cellular cytoplasm becomes possible. Furthermore, and for the first time, the inherent advantages in resolution of the near-field images provides highly specific details of the location of a GFP fusion protein on a bacterial cell surface.
authors.affiliation: Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, U.K. patrick.gunning@bbsrc.ac.uk
authors.forename: A P
authors.initials: AP
authors.lastname: Gunning
authors.forename: R J M
authors.initials: RJ
authors.lastname: Bongaerts
authors.forename: A R
authors.initials: AR
authors.lastname: Kirby
authors.forename: J C D
authors.initials: JC
authors.lastname: Hinton
authors.forename: V J
authors.initials: VJ
authors.lastname: Morris
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D018625:Microscopy, Atomic Force; D008856:Microscopy, Fluorescence / Q000379:methods*; D020527:Microscopy, Scanning Probe / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012760:Shigella / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; C058483:virG protein, Shigella flexneri; D049452:Green Fluorescent Proteins