title: Identification of gyrB and rpoB gene mutations and differentially expressed proteins between a novobiocin-resistant Aeromonas hydrophila catfish vaccine strain and its virulent parent strain.
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. 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 Aeromonas hydrophila infection.
authors: Aquatic Animal Health Research Unit, USDA-ARS, 990 Wire Road, Auburn, AL 36830, USA. Julia.Pridgeon@ars.usda.gov
authors: J W
authors: JW
authors: Pridgeon
authors: M
authors: M
authors: Yildirim-Aksoy
authors: P H
authors: PH
authors: Klesius
authors: K
authors: K
authors: Kojima
authors: J A
authors: JA
authors: Mobley
authors: K K
authors: KK
authors: Srivastava
authors: P G
authors: PG
authors: Reddy
mesh_terms: D016980:Aeromonas hydrophila / Q000187:drug effects / Q000235:genetics* / Q000276:immunology / Q000472:pathogenicity; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics* / Q000276:immunology; D001428:Bacterial Vaccines / Q000235:genetics / Q000276:immunology; D015153:Blotting, Western; D002397:Catfishes / Q000382:microbiology; D027081:DNA Gyrase / Q000235:genetics* / Q000276:immunology; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D024881:Drug Resistance, Bacterial; D005393:Fish Diseases / Q000382:microbiology*; D005634:Fructose-Bisphosphate Aldolase / Q000235:genetics; D016905:Gram-Negative Bacterial Infections / Q000382:microbiology / Q000662:veterinary*; D009154:Mutation; D009675:Novobiocin / Q000494:pharmacology*
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D001428:Bacterial Vaccines; D009675:Novobiocin; D012321:DNA-Directed RNA Polymerases; C105078:RNA polymerase beta subunit; D005634:Fructose-Bisphosphate Aldolase; D027081:DNA Gyrase
keywords: Aeromonas hydrophila; Immunogen; Mass spectrometry; Virulence factor