Identification and evaluation of an outer membrane protein OmpU from a pathogenic Vibrio harveyi isolate as vaccine candidate in turbot (Scophthalmus maximus)

Lett Appl Microbiol. 2011 Jul;53(1):22-9. doi: 10.1111/j.1472-765X.2011.03062.x. Epub 2011 May 12.

Abstract

Aims: The main aims of this study were to clone and express a new outer membrane protein U (OmpU) from a pathogenic Vibrio harveyi SF-1 and investigate its immune efficiency as a vaccine candidate against V. harveyi infection in turbot (Scophthalmus maximus).

Methods and results: In this study, a new gene, ompU was cloned from the genomic DNA of pathogenic V. harveyi SF-1. The ompU gene encoded a 35 kDa protein, which was purified by Ni-NTA His-Bind Resin column. A DNA vaccine was constructed by inserting ompU gene into pEGFP-N1 plasmid. Turbot were injected intramuscularly with the purified OmpU protein and the recombinant pEGFP-N1/ompU plasmid, respectively. The fish vaccinated with the purified OmpU protein were completely protected with a relative per cent of survival (RPS) of 100% against pathogenic V. harveyi infection. Efficient protection was also found in the pEGFP-N1/ompU vaccinated group, with a RPS of 51.4%. Significant specific antibody responses were detected in the vaccinated turbot by indirect enzyme-linked immunosorbent assay.

Conclusions: A new OmpU was cloned and expressed. Both OmpU protein vaccine and DNA vaccine showed good immune protections in turbot.

Significance and impact of the study: The OmpU was identified to be a new effective vaccine candidate and could be used as subunit vaccine and DNA vaccine for disease control caused by pathogenic V. harveyi.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Outer Membrane Proteins / immunology
  • Bacterial Vaccines / immunology*
  • Cloning, Molecular
  • Enzyme-Linked Immunosorbent Assay
  • Fish Diseases / immunology*
  • Fish Diseases / microbiology
  • Fish Diseases / prevention & control*
  • Flatfishes*
  • Gene Expression
  • Green Fluorescent Proteins / genetics
  • Plasmids
  • Transcription, Genetic
  • Vaccines, DNA / immunology
  • Vibrio / chemistry*
  • Vibrio / genetics*
  • Vibrio / physiology
  • Vibrio Infections / immunology
  • Vibrio Infections / prevention & control
  • Vibrio Infections / veterinary*

Substances

  • Bacterial Outer Membrane Proteins
  • Bacterial Vaccines
  • Vaccines, DNA
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins