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Prokaryotic expression and immunoassay of grass carp reovirus capsid VP6 protein

文献类型： 外文期刊

作者： Zhou, Y. 1 ; Fan, Y. D. 1 ; Zeng, L. B. 1 ; Ma, J. 1 ;

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

关键词： grass carp reovirus;VP6 gene/protein;polyclonal antibody;Western blot analysis;immunofluorescence staining

期刊名称：ACTA VIROLOGICA （ 影响因子：1.162； 五年影响因子：1.255 ）

ISSN： 0001-723X

年卷期： 2013 年 57 卷 4 期

页码：

收录情况： SCI

摘要： Grass carp reovirus (GCRV) of the genus Aquareovirus and the family Reoviridae causes a severe hemorrhagic disease in grass carp fingerlings in China. GCRV genome is composed of 11 double-stranded RNA segments, of which segment 8 encodes the major core capsid protein VP6. In this study, the VP6 gene following an RT-PCR-amplification from the GCRV 104 strain was cloned into an expression vector pET-32a to obtain pET-32(a)-VP6. The VP6 protein was expressed in Escherichia coli BL21 as a fusion protein of 64 kDa. After purification with the HisLink Spin Protein Purification System the VP6 protein was used to raise a specific polyclonal antibody in Balb/c mice. Presence of VP6 protein was proved in bacterial lysates containing VP6 fusion protein by Western blot analysis and in GCRV-infected CIK cells by immunofluorescent staining using polyclonal antibody. These results may be helpful in further studies of interactions between GCRV and cells and in preparation of an engineered vaccine against GCRV.

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