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Characterization of the outer membrane proteome of Francisella noatunensis subsp. orientalis
Author(s) -
Shahin K.,
Thompson K.D.,
Inglis N.F.,
Mclean K.,
RamirezParedes J.G.,
Monaghan S.J.,
Hoare R.,
Fontaine M.,
Metselaar M.,
Adams A.
Publication year - 2018
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/jam.13918
Subject(s) - biology , tilapia , proteome , nile tilapia , microbiology and biotechnology , bacterial outer membrane , western blot , virulence , oreochromis , escherichia coli , biochemistry , gene , fishery , fish <actinopterygii>
Aims The aims of the current study were to characterize the outer membrane proteins ( OMP s) of Francisella noatunensis subsp. orientalis ( Fno ) STIR ‐ GUS ‐F2f7, and identify proteins recognized by sera from tilapia, Oreochromis niloticus , (L) that survived experimental challenge with Fno . Methods and Results The composition of the OMP s of a virulent strain of Fno ( STIR ‐ GUS ‐F2f7), isolated from diseased red Nile tilapia in the United Kingdom, was examined. The sarcosine‐insoluble OMPs fraction was screened with tilapia hyperimmune sera by western blot analysis following separation of the proteins by 1D SDS ‐ PAGE . Liquid chromatography‐electrospray ionization‐tandem mass spectrometry ( LC ‐ ESI ‐ MS / MS ) was used to identify the various proteins present in the OMP profile. Two hundred and thirty‐nine proteins were identified, of which 44 were found in the immunogenic band recognized by the tilapia hyperimmune serum. In silico analysis was performed to predict the function and location of the OMP s identified by MS . Conclusions Using a powerful proteomic‐based approach in conjugation with western immunoblotting, proteins comprising the outer membrane fraction of Fno STIR ‐ GUS ‐F2f7 were identified, catalogued and screened for immune recognition by tilapia sera. Significance and Impact of the Study The current study is the first report on the characterization of Fno ‐ OMP s. The findings here provide preliminary data on bacterial surface proteins that exist in direct contact with the host's immune defences during infection and offer an insight into the pathogenesis of Fno .