Bombyx mori nucleopolyhedrovirus (BmNPV) orf25 gene was characterized for the first time. The coding sequenceof Bin25 was amplified and subcloned into the prokaryotic expression vector pGEX-4T-2 to produce glutathioneS-transferase-tagged fusion protein in the BL21 (DE3) cells. The GST-Bm25 fusion protein was expressed efficiently afterinduction with IPTG. The purified fusion protein was used to immunize New Zealand white rabbits to prepare polyclonal an-tibody. Temporal expression analysis revealed a 30-kDa protein, which was detected beginning 24 hours post-infection using apolyclonal antibody against GST-Bm25 fusion protein. The transcript of Bm25 was detected by RT-PCR at 18-72 h p.i. In con-clusion, the available data suggest that Bm25 encodes a 30kDa protein expressed in the late stage of infection cycle.%首次对家蚕核型多角体orf25基因进行了描述.扩增Bm25基因,亚克隆到原核表达载体pGEX-4T-2,在大肠杆菌BL21(DE3)中表达含有GST标签的融合蛋白.IPTG诱导后高效表达GST-Bm25融合蛋白.纯化的融合蛋白免疫新西兰大白兔制备多克隆抗体.利用制备的抗GST-Bm25融合蛋白的多克隆抗体进行表达时相分析显示:24 h p.i.检测到30 kDa的蛋白条带.RT-PCR方法,在18-72 h p.i 检测到Bm25基因的转录本.结论:以上数据表明Bm25基因编码一晚期表达的30kDa蛋白.
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