Location: Aquatic Animal Health ResearchTitle: Construction, expression and characterization of eleven putative flagellar apparatus genes of Aeromonas hydrophila AL09-73) Author
Submitted to: Journal of Fish Diseases
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 9/6/2011
Publication Date: 11/1/2012
Publication URL: http://handle.nal.usda.gov/10113/56371
Citation: Yeh, H., Klesius, P.H. 2012. Construction, expression and characterization of eleven putative flagellar apparatus genes of Aeromonas hydrophila AL09-73. Journal of Fish Diseases. 35:853-860. Interpretive Summary: Aeromonas hydrophila is ubiquitous in aquatic environments and is responsible for many human and fish diseases. Recent outbreaks of aeromonad hemorrhagic septicemia in channel catfish in the State of Alabama have caused huge economic losses to the producers. Development of appropriate means for this disease control is in great demand and will minimize losses caused by this microorganism. Eleven Aeromonas hydrophila flagellar proteins were amplified, expressed and characterized in an E. coli bacterial expression system. These recombinant proteins can be used for the subunit vaccine development and studying catfish antibody dynamics after infection. This study is related to the goal of ARS National Program 106-Aquaculture, Component 4C-Prevention of Diseases.
Technical 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.