鳜传染性脾肾坏死病毒ORF093基因的克隆、表达及其重组蛋白的免疫原性分析
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中国水产科学研究院 珠江水产研究所, 农业部渔药创制重点实验室, 广东 广州 510380

作者简介:

付小哲(1978), 女, 助理研究员, 从事水生动物病害防治研究. E-mail: fuxiaozhe-1998@163.com

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S941

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国家自然科学基金资助项目(31202032); 国家科技支撑计划项目(2012BAD25B02); 广东省自然科学基金资助项目(7004728).


Cloning and expression of ORF093 gene and the immune effect of its recombinant protein of infectious spleen and kidney necrosis virus in Siniperca chuatsi
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Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Key laboratory of Fishery Drug Development, Ministry of Agriculture, Guangzhou 510380, China

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    摘要:

    (infectious spleen and kidney necrosis virus, ISKNV)因。采用可以在大肠杆菌中以包涵体形式存在ISKNV重组的攻击具有保护作用组平均相对保护率为蛋白具有一定的免疫原性候选疫苗抗原和治疗血清制备抗原。本研究通过中和实验及免疫保护实验对重组

    Abstract:

    gene from infectious spleen and kidney necrosis virus (ISKNV) isolated from mandarinfish,, was amplified by PCR, and then cloned into the prokaryotic expression plasmid pET32a(+). The recombinant 093 protein expression was induced by IPTG and purified by Ni-NTA affinity chromatograph. Polyclonal antibodies were raised in rabbits against the purified protein and the reaction of the antibody was confirmed by western blotting using the purified protein and the spleen and kidney of healthy and diseased mandarin fish. The immunogenicity of 093 protein was investigated by neutralization test and immune protection test. The results showed that the recombinant 093 protein was found as the inclusion bodies in . Western blotting of rabbit sera against recombinant 093 protein and the spleen and kidney of diseased mandarin fish showed a protein recognition. The antiserum could neutralize ISKNV infection and could provide fish 10-days-long 100% protection. The recombinant 093 protein could protect mandarinfish against virulent challenge with ISKNV and the average RPS of 093 group was 45.3%. The results indicated that the recombinant 093 protein had It could be a potential vaccine candidate and antigen candidate for antiserum treatment.

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付小哲,李宁求,彭媛媛,林强,石存斌,黄志斌,吴淑勤.鳜传染性脾肾坏死病毒ORF093基因的克隆、表达及其重组蛋白的免疫原性分析[J].中国水产科学,2013,20(2):427-433
FU Xiaozhe, LI Ningqiu, PENG Yuanyuan, LIN Qiang, SHI Cunbin, HUANG Zhibin, WU Shuqin. Cloning and expression of ORF093 gene and the immune effect of its recombinant protein of infectious spleen and kidney necrosis virus in Siniperca chuatsi[J]. Journal of Fishery Sciences of China,2013,20(2):427-433

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  • 在线发布日期: 2013-04-08
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