Letters in Applied Microbiology · 2011 · 47 citations · 35 references
Veterinary MedicineMicrobiologyVeterinary MicrobiologyHost-pathogen InteractionsVaccine BiotechnologyVaccinologyDna VaccinesPathogen Virulence FactorsRecombinant Protein VaccinesVaccine Antigen DesignVaccine Antigen DiscoveryVeterinary VaccinesAquaculture VaccinesBacterial Fish VaccinesOuter Membrane ProteinsProtective Antigen IdentificationPurification Of Vaccine AntigensVaccine Candidate AntigensVaccine Candidate ScreeningVaccine Characterization
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.
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Molecular cloning: A laboratory manual
Prescott L. Deininger · Analytical Biochemistry · 1990 · 86.2K citations