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Vaccine. 2007 Mar 1;25(11):2095-102. Epub 2006 Dec 4.

Immunological characterization of a bacterial protein isolated from salmonid fish naturally infected with Piscirickettsia salmonis.

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  • 1Laboratorio de Genética e Inmunología Molecular, Instituto de Biología, Facultad de Ciencias Básicas y Matemáticas, Pontificia Universidad Católica de Valparaíso, Avenida Brasil 2950, Valparaíso, Chile. smarshal@ucv.cl

Abstract

The Salmon Rickettsia syndrome (SRS) remains a major infectious disease in the Chilean aquaculture. A limited number of Piscirickettsia salmonis proteins have been characterized so far for their use as potential candidates for vaccines studies. In this study, we identified and expressed a highly immunogenic protein of P. salmonis extracted by selective hydrophobicity from crude-cell macerates of naturally infected salmonid fish. One and two-D PAGE gels followed by Western blot analysis with a battery of polyclonal anti-P. salmonis antibodies have allowed the isolation of the target protein. Basic local alignment search (BLAST) done after partial sequencing of the pure protein identified it as a member of the heat-shock protein (HSP) family of prokaryotes. The protein, named ChaPs, was cloned as a single open reading frame encoding 545 amino acid residues with a predicted molecular mass of 57.3 kDa. The amplicon representing the entire novel gene was expressed in vitro in different heterologous systems: the PurePro Caulobacter crescentus expression system from where most of the characterization was attained, and also in the Escherichia coli BL-21 CodonPlus model for commercially potential purposes. The immunologic potential of ChaPs was determined with serum from naturally infected fish.

PMID:
17250933
[PubMed - indexed for MEDLINE]
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