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United States Department of Agriculture

Agricultural Research Service

Research Project: Integrated Aquatic Animal Health Strategies

Location: Aquatic Animal Health Research

Title: Molecular characterization of a novobiocin-resistant Aeromonas hydrophila catfish vaccine strain compared to its virulent parent strain

Authors
item Wei Pridgeon, Yuping
item Aksoy, Mediha -
item Klesius, Phillip

Submitted to: Aquaculture America Conference
Publication Type: Proceedings
Publication Acceptance Date: August 30, 2013
Publication Date: February 9, 2014
Citation: Wei Pridgeon, Y., Aksoy, M., Klesius, P.H. 2014. Molecular characterization of a novobiocin-resistant Aeromonas hydrophila catfish vaccine strain compared to its virulent parent strain. Proceedings Aquaculture America 2014. p. 424.

Technical Abstract: A total of 10 and 13 missense mutations were found in the deduced gyrB and rpoB proteins, respectively, between avirulent AH11NOVO vaccine strain and its virulent parent strain AH11P. SDS-PAGE revealed that six proteins bands were significantly over-expressed in AH11NOVO whereas five bands were significantly over-expressed in AH11P (Figure 1). Mass spectrometry identified seven proteins from the over-expressed AH11NOVO gel bands and five proteins from the over-expressed AH11P gel bands. QPCR confirmed that all 12 genes corresponding to the proteins identified by mass spectrometry were significantly over-expressed in AH11NOVO or AH11P. When AH11NOVO proteins were subjected to Western blot analysis, 13 protein bands exhibited significantly stronger reactivity with hyper-immune catfish sera. Fifteen proteins were identified from immunogenic protein bands, including six (formate acetyltransferase, chaperone htpG, transketolase, ATP synthase subunit alpha, asparagine-tRNA ligase, and serine hydroxymethyltransferase) that were over-expressed in AH11NOVO proteins and three (elongation factor G, class II fructose-bisphosphate aldolase, and a putative uncharacterized 23 kDa protein) that were over-expressed in AH11P. In addition, the following six proteins were also identified from the immunogenic protein bands: pyruvate dehydrogenase E1 component, ATP synthase subunit beta, ribose-phosphate pyrophosphokinase, glyceraldehyde-3-phosphate dehydrogenase, 50S ribosomal L10, and 50S ribosomal L15. Our results might provide insights on how to develop novel efficacious vaccine against A. hydrophila infection.

Last Modified: 12/21/2014
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