Journal article
Comparative proteomic analysis of virulent and rifampicin-attenuated Flavobacterium psychrophilum
Journal of fish diseases, Vol.35(7), pp.529-539
07/2012
Handle:
https://hdl.handle.net/2376/112782
PMID: 22607560
Abstract
Flavobacterium psychrophilum is the aetiologic agent of bacterial coldwater disease and rainbow trout fry syndrome. In this study, we compared a wild-type strain (CSF 259-93) with a rifampicin-resistant strain and virulence-attenuated strain of F. psychrophilum (CSF 259-93B.17). The attenuated strain harboured a mutation in the rpoB gene consistent with resistance to rifampicin. Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and mass spectrometry demonstrated an altered proteome with eight proteins characteristic for the parent strain and six that were unique to the attenuated strain. Immunoblotting with a diagnostic monoclonal antibody (FL-43) identified a putative antigen (FP1493) that was subsequently cloned, expressed as a recombinant protein and confirmed as recognized by FL-43. 2D-PAGE, immunoblotting with rainbow trout, Oncorhynchus mykiss (Walbaum), convalescent antisera and mass spectrometry of bacterial whole-cell lysates revealed several uniquely expressed immunoreactive proteins including FP1493. An FP1493 recombinant subunit vaccine was tested, but did not provide protection against challenge with the CSF259-93 strain. While the exact mechanism responsible for altered protein synthesis and attenuation of CSF 259-93B.17 is still unknown, the differentially expressed immunoreactive proteins are a valuable resource to develop subunit vaccines and to identify proteins that are potentially involved in disease.
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Details
- Title
- Comparative proteomic analysis of virulent and rifampicin-attenuated Flavobacterium psychrophilum
- Creators
- K Gliniewicz - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 99164-7040, USAK P PlantS E LaPatraB R LaFrentzK CainK R SnekvikD R Call
- Publication Details
- Journal of fish diseases, Vol.35(7), pp.529-539
- Academic Unit
- Center for Reproductive Biology; Veterinary Microbiology and Pathology, Department of; Paul G. Allen School for Global Animal Health
- Publisher
- England
- Identifiers
- 99900548248501842
- Language
- English
- Resource Type
- Journal article