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Construction, expression and characterization of 11 putative flagellar apparatus genes of Aeromonas hydrophila AL09-73

Author:
Yeh, H-Y, Klesius, P. H.
Source:
Journal of fish diseases 2012 v.35 no.11 pp. 853
ISSN:
1365-2761
Subject:
Aeromonas hydrophila, Ictalurus punctatus, animal pathogenic bacteria, antibodies, antigen-antibody reactions, aquatic environment, bacterial antigens, bacterial infections, bacterial proteins, databases, fish diseases, flagellum, gene expression, genes, mice, molecular cloning, polyacrylamide gel electrophoresis, polymerase chain reaction, recombinant proteins
Abstract:
Aeromonas hydrophila is ubiquitous in aquatic environments worldwide and is responsible for many human and fish diseases. The genome sequence of A. hydrophila is available in the GenBank database. Our ultimate goal is to develop the whole genome protein arrays of A. hydrophila. In this study, we PCR amplified all flagellar genes except the flgH, flhA, flhD and fliF genes. Eleven flagellar genes were randomly selected for further cloning and expression analysis. Except the flgN protein, all selected flagellar genes were expressed. The recombinant proteins migrated to the expected size positions on the Coomassie-stained SDS-PAGE gels, and were reactive to a mouse anti-His tag antibody, suggesting that these proteins were expressed as fusion proteins. In addition, serum from channel catfish experimentally infected with A. hydrophila was reacted strongly to the flgE, flgI, flgL, fliH, fliL and fliM, but not flgD, flgJ, flhF and fliN proteins, implying that the accessibility of the latter four proteins for antigen processing during the infection is limited. The functions of these genes/proteins are under investigation.
Agid:
56371
Handle:
10113/56371