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Vaccination of plasmid DNA encoding ORF81 gene of CJ strains of KHV provides protection to immunized carp

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Abstract

In order to construct the recombinant plasmid of pIRES-ORF81, the nucleic acid isolated from Koi herpes virus-CJ (KHV-CJ) strains was used as a template to insert the ORF81 gene fragments amplified by PCR into the pIRES-neo, a kind of eukaryotic expression vector. Using Western blotting analysis, it was verified that ORF81 gene protein can be expressed correctly by pIRES-ORF81, after MFC cells were transfected. The recombinant plasmid pIRES-ORF81 was set into three immunization dose gradients: 1, 10, and 50 μg/carp. Empty plasmid group, PBS group, and blank control group were set simultaneously. Giving intramuscular injections to healthy carps with an average body mass of 246 ± 20 g, indirect ELISA was used to regularly determine antibody levels after three times immunization injection. Neutralizing antibodies were detected by neutralization assay. The results of inoculation tests showed that the pIRES-ORF81 recombinant plasmid can induce the production of carp-specific antibodies. The differences of immune effect between the three different doses of immune gradients were not significant (P > 0.05), but they can induce the production of neutralizing antibodies. After 25 d of inoculation, carp mortality of pIRES-neo empty vector treatment groups was 85%, while the carp mortality of eukaryotic expression recombinant plasmid pIRES-ORF81 injected with three different doses of immune gradients was 20, 17.5, and 12.5%, respectively. Differences in comparison to the control group were highly significant (P < 0.01). However, histopathological section of immunohistochemistry organization revealed no significant changes. It demonstrated that the DNA vaccine pIRES-ORF81 constructed in the experiment displayed a good protective effect against KHV, which had the potential to industrial applications.

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Acknowledgements

This study was funded by the National Natural Science Foundation of China (No. 30671621), supported by the World Bank project in Jilin Province (No. 2011-YS13) and industrial technology research and exploitation project of Jilin Province (No. 2011002–1).

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Authors and Affiliations

  1. College of Animal Science and Technology, Key Laboratory of Animal Production and Safety of Ministry of Education, Jilin Agricultural University, Changchun, 130118, China

    Jingxiang Zhou, Qiuju Wang, Xia Zhu, Xingwei Li, Wenliang Lv & Dongming Zhang

  2. College of Animal Science and Technology, Inner Mongolia University for Nationalities, Tongliao, 028000, China

    Jiangdong Xue

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  1. Jingxiang Zhou
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  2. Jiangdong Xue
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  3. Qiuju Wang
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  4. Xia Zhu
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  5. Xingwei Li
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  6. Wenliang Lv
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  7. Dongming Zhang
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Correspondence to Dongming Zhang.

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Editor: T. Okamoto

Jingxiang Zhou and Jiangdong Xue contributed equally to this work.

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Zhou, J., Xue, J., Wang, Q. et al. Vaccination of plasmid DNA encoding ORF81 gene of CJ strains of KHV provides protection to immunized carp. In Vitro Cell.Dev.Biol.-Animal 50, 489–495 (2014). https://doi.org/10.1007/s11626-014-9737-2

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  • DOI: https://doi.org/10.1007/s11626-014-9737-2

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