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将含有斜带石斑鱼Epinephelus coioides神经坏死病毒(orange-spotted grouper nervous necrosis virus,OGNNV)主衣壳蛋白(major capsid protein,MCP)基因的重组表达质粒载体pET32a_MCP转化到大肠杆菌BL21(DE3)中进行融合表达,经SDS_PAGE分析和Western_blot鉴定,证实了重组大肠杆菌融合表达了斜带石斑鱼神经坏死病毒主衣壳蛋白。表达的融合蛋白主要以不可溶的包涵体形式存在,提取的包涵体中融合蛋白含量占60%以上,经柱层析纯化蛋白,纯化度达90%以上。
The recombinant plasmid pET32a_MCP containing the major capsid protein (MCP) gene of Epinephelus coioides necrosis virus (OGNNV) was transformed into E.coli BL21 (DE3) After SDS-PAGE analysis and Western_blot identification, we confirmed that the recombinant Escherichia coli fused and expressed the main capsid protein of the serrated necrotic necrosis virus. The expressed fusion protein mainly existed in the form of insoluble inclusion bodies. The content of the fusion protein in the inclusion body accounted for more than 60%. The purified protein was purified by column chromatography and the purity was over 90%.