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Protective immunity of a Pichia pastoris expressed recombinant iridovirus major capsid protein in the Chinese giant salamander, Andrias davidianus

文献类型： 外文期刊

作者： Zhou, Yong 1 ; Fan, Yuding 1 ; LaPatra, Scott E. 3 ; Ma, Jie 1 ; Xu, Jin 1 ; Meng, Yan 1 ; Jiang, Nan 1 ; Zeng, Lingbing 1 ;

作者机构： 1.Chinese Acad Fishery Sci, Yangtze River Fisheries Res Inst, Wuhan 430223, Peoples R China

2.Huazhong Agr Univ, Coll Fisheries, Wuhan 430071, Peoples R China

3.Clear Springs Foods Inc, Div Res, Buhl, ID 83316 USA

关键词： Chinese giant salamander;Iridovirus;MCP;Pichia pastoris expression;Protective immunity

期刊名称：VACCINE （ 影响因子：3.641； 五年影响因子：3.816 ）

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收录情况： SCI

摘要： The major capsid protein (MCP) is the main immunogenic protein of iridoviruses, that has been widely used as an immunogen in vaccination trials. In this study, the codon-optimized giant salamander iridovirus (GSIV) MCP gene (O-MCP) was synthesized and cloned into a pPICZ alpha B vector for secretory expression in the methylotrophic yeast Pichia pastoris after methanol induction. The expression of the O-MCP protein was detected by the Bradford protein assay, SDS-PAGE, Western blotting and electron microscopy. The Bradford protein assay indicated that the concentration of the O-MCP expressed was about 40 mu g/ml in culture supernatants. SOS-PAGE analysis revealed that the O-MCP had a molecular weight of about 66 kDa and reacted with a His-specific MAb that was confirmed by Western blotting. Electron microscopy observations revealed that the purified O-MCP could self-assemble into virus-like particles. Healthy giant salamanders were vaccinated by intramuscular injection with the O-MCP antigen at a dose of 20 mu g/individual. The numbers of erythrocytes and leukocytes in the peripheral blood of immunized Chinese giant salamanders increased significantly at day 3 and reached a peak at day 5 post-immunization. Meanwhile, the differential leukocyte counts of monocytes and neutrophils increased significantly at day 5 post-immunization compared to that of the control group. The percentage of lymphocytes was 71.33 +/- 13.57% at day 21 post-immunization.

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