Confirming the Functional Roles of the Ortholog Genes ahpB of Aeromonas hydrophila and hap of Aeromonas salmonicida

Presenters

Disciplines

Bacteriology | Genetics | Molecular Biology | Molecular Genetics | Other Immunology and Infectious Disease | Pathogenic Microbiology

Abstract (300 words maximum)

Aeromonas salmonicida, a globally-distributed, aquatic bacterium is a fish pathogen with high mortality rates causing severe hemorrhaging. Open wounds, warm waters, and crowded habitats allow A. salmonicida to spread quickly in a population, costing the fishing industry over $40 million annually. Aeromonas hydrophila, a related species, is a similar fish pathogen; however, A. hydrophila also infects humans, leading to clinical diseases including gastroenteritis and rapid flesh destruction.

Relative to the A. hydrophila genome, A. salmonicida has undergone substantial changes based on its adaptation to salmonid hosts, providing insights into the mechanisms used for infection. However, they maintain similar genes such as hap of A. salmonicida and ahpB of A. hydrophila. The gene hap is reported to encode a hemagglutinin/protease. Hemagglutinin is a factor in the clumping of red blood cells (RBCs). In A. hydrophila, the gene with highest homology to hap is ahpB, which encodes an elastin degrading enzyme called elastase. Elastin is the extracellular protein enabling elastic tissues like skin to stretch. Using an elastin-orcein assay, we have shown that both A. salmonicida (unexpected) and A. hydrophila (expected) exhibit elastase activity. Therefore, this study aims to confirm the functional roles of the proteins encoded by hap and ahpB to understand their role in the progression of disease, and if they should be targeted to reduce disease severity.

This is accomplished by cloning the hap and ahpB genes into strains of E. coli, followed by induction of genes. Elastin-orcein and hemagglutination assays test for new functionalities in the transformed E. coli. The protein product is later purified and analyzed.

Academic department under which the project should be listed

CSM - Molecular and Cellular Biology

Primary Investigator (PI) Name

Dr. Donald McGarey

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Confirming the Functional Roles of the Ortholog Genes ahpB of Aeromonas hydrophila and hap of Aeromonas salmonicida

Aeromonas salmonicida, a globally-distributed, aquatic bacterium is a fish pathogen with high mortality rates causing severe hemorrhaging. Open wounds, warm waters, and crowded habitats allow A. salmonicida to spread quickly in a population, costing the fishing industry over $40 million annually. Aeromonas hydrophila, a related species, is a similar fish pathogen; however, A. hydrophila also infects humans, leading to clinical diseases including gastroenteritis and rapid flesh destruction.

Relative to the A. hydrophila genome, A. salmonicida has undergone substantial changes based on its adaptation to salmonid hosts, providing insights into the mechanisms used for infection. However, they maintain similar genes such as hap of A. salmonicida and ahpB of A. hydrophila. The gene hap is reported to encode a hemagglutinin/protease. Hemagglutinin is a factor in the clumping of red blood cells (RBCs). In A. hydrophila, the gene with highest homology to hap is ahpB, which encodes an elastin degrading enzyme called elastase. Elastin is the extracellular protein enabling elastic tissues like skin to stretch. Using an elastin-orcein assay, we have shown that both A. salmonicida (unexpected) and A. hydrophila (expected) exhibit elastase activity. Therefore, this study aims to confirm the functional roles of the proteins encoded by hap and ahpB to understand their role in the progression of disease, and if they should be targeted to reduce disease severity.

This is accomplished by cloning the hap and ahpB genes into strains of E. coli, followed by induction of genes. Elastin-orcein and hemagglutination assays test for new functionalities in the transformed E. coli. The protein product is later purified and analyzed.