title: Development of a Recombinant Protein Vaccine Based on Cell-Free Protein Synthesis for Sevenband Grouper Epinephelus septemfasciatus Against Viral Nervous Necrosis.
abstract: Sevenband grouper, Epinephelus septemfasciatus, is becoming an important aquaculture species in Korea. However, viral nervous necrosis disease is a large problem causing mass mortality in sevenband grouper aquaculture. Recombinant protein vaccines are one of the best methods to reduce these economic losses. However, the cell-based expression method mainly produces inclusion bodies and requires additional procedures. In this study, we expressed a recombinant viral coat protein of sevenband grouper nervous necrosis virus (NNV) using a cell-free protein synthesis system. The purified recombinant NNV coat protein (rNNV-CP) was injected into sevenband grouper at different doses followed by a NNV challenge. Nonimmunized fish in the first trial (20 μg/fish) began to die 5 days post-challenge and reached 70% cumulative mortality. In contrast, immunized fish also starting dying 5 days postchallenge but lower cumulative mortality (10%) was observed. Cumulative morality in the second trial with different doses (20, 4, and 0.8 μg/fish) was 10%, 40%, and 50%, respectively. These results suggest that rNNV-CP can effectively immunize sevenband grouper depending on the dose administered. This study provides a new approach to develop a recombinant vaccine against NNV infection for sevenband grouper.
authors.affiliation: Department of Aqualife Medicine, College of Fisheries and Ocean Science, Chonnam National University, Yeosu 550-749, Republic of Korea.
authors.forename: Jong-Oh
authors.initials: JO
authors.lastname: Kim
authors.affiliation: Department of Aqualife Medicine, College of Fisheries and Ocean Science, Chonnam National University, Yeosu 550-749, Republic of Korea.
authors.forename: Jae-Ok
authors.initials: JO
authors.lastname: Kim
authors.affiliation: Department of Aqualife Medicine, College of Fisheries and Ocean Science, Chonnam National University, Yeosu 550-749, Republic of Korea.
authors.forename: Wi-Sik
authors.initials: WS
authors.lastname: Kim
authors.affiliation: Department of Aqualife Medicine, College of Fisheries and Ocean Science, Chonnam National University, Yeosu 550-749, Republic of Korea.
authors.forename: Myung-Joo
authors.initials: MJ
authors.lastname: Oh
mesh_terms: D000818:Animals; D001492:Bass; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology*; D005393:Fish Diseases / Q000517:prevention & control*; D007723:Korea; D030381:Nodaviridae / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D016019:Survival Analysis; D016896:Treatment Outcome; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D014765:Viral Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D014777:Virus Diseases / Q000517:prevention & control / Q000662:veterinary*
chemical_list: D036022:Capsid Proteins; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; D014765:Viral Vaccines
keywords: cell free protein synthesis (CFPS); nervous necrosis virus (NNV); recombinant protein; sevenband grouper; viral nervous necrosis (VNN)