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家蚕核型多角体病毒的orf25在感染周期中编码一种30kDa的晚期蛋白。

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

作者信息

Wang Haiyan, Chen Keping, Guo Zhongjian, Yao Qin

机构信息

Institute of Life Sciences, Jiangsu University, Zhenjiang 212013, China.

出版信息

Wei Sheng Wu Xue Bao. 2008 Feb;48(2):247-51.

Abstract

Bombyx mori nucleopolyhedrovirus (BmNPV) orf25 gene was characterized for the first time. The coding sequence of Bm25 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 Bm25 encodes a 30kDa protein expressed in the late stage of infection cycle.

摘要

首次对家蚕核型多角体病毒(BmNPV)的orf25基因进行了表征。扩增了Bm25的编码序列,并将其亚克隆到原核表达载体pGEX-4T-2中,以在BL21(DE3)细胞中产生谷胱甘肽S-转移酶标记的融合蛋白。用IPTG诱导后,GST-Bm25融合蛋白高效表达。纯化的融合蛋白用于免疫新西兰白兔以制备多克隆抗体。时序表达分析显示出一种30 kDa的蛋白质,使用针对GST-Bm25融合蛋白的多克隆抗体在感染后24小时开始检测到该蛋白质。在感染后18 - 72小时通过RT-PCR检测到Bm25的转录本。总之,现有数据表明Bm25编码一种在感染周期后期表达的30 kDa蛋白质。

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