家蚕核型多角体病毒orf25基因在病毒感染过程中编码一个30kDa的晚期蛋白(英文)  

Bombyx mori nucleopolyhedrovirus orf25 encodes a 30kDa late protein in the infection cycle

作　　者：王海燕[1] 陈克平[1] 郭忠建[1] 姚勤[1] 

机构地区：[1]江苏大学生命科学研究院,镇江212013

出　　处：《微生物学报》2008年第2期247-251,共5页Acta Microbiologica Sinica

基　　金：国家自然科学基金(30370773);国家"973项目"(2005CB121000)~~

摘　　要：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:24h 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。Ab Bombyx mori nucleopolyhedrovirus （BmNPV） orf25 gene was characterized for the first time. The coding sequence of Bin25 was amplified and subcloned into the prokaryotic expression vector pGEX-4T-2 to produce glutathione S-transferase-tagged fusion protein in the BL21 （DE3） cells. The GST-Bm25 fusion protein was expressed efficiently after induction with IPTG. The purified fusion protein was used to immunize New Zealand white rabbits to prepare polyclonal antibody. Temporal expression analysis revealed a 30-kDa protein, which was detected beginning 24 hours post-infection using a polyclonal antibody against GST-Bm25 fusion protein. The transcript of Bm25 was detected by RT-PCR at 18-72 h p.i. In conclusion, the available data suggest that Bin25 encodes a 30kDa protein expressed in the late stage of infection cycle.

关 键 词：BMNPV Bin25 表达时相 RT-PCR 多克隆抗体 

分 类 号：Q939.4[生物学—微生物学]