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家蚕核型多角体病毒ORF56编码一种源自包涵体的病毒蛋白,对出芽病毒的产生并非必需。

Bombyx mori nucleopolyhedrovirus ORF56 encodes an occlusion-derived virus protein and is not essential for budded virus production.

作者信息

Xu Hai-Jun, Yang Zhang-Nv, Zhao Jin-Fang, Tian Cai-Hong, Ge Jun-Qing, Tang Xu-Dong, Bao Yan-Yuan, Zhang Chuan-Xi

机构信息

Institute of Insect Science, Zhejiang University, Hangzhou 310029, PR China.

出版信息

J Gen Virol. 2008 May;89(Pt 5):1212-1219. doi: 10.1099/vir.0.83633-0.

Abstract

Bombyx mori nucleopolyhedrovirus ORF56 (Bm56) is a baculovirus core gene that is highly conserved in all baculoviruses that have had their genomes sequenced to date. Its transcripts in BmNPV-infected cells could be detected from 12 h post-infection (p.i.) and the encoded protein could be detected at 16 h p.i. by using a polyclonal antibody against glutathione S-transferase-Bm56 fusion protein. Western blot analysis showed that Bm56 is a structural component of the occlusion-derived virus nucleocapsid. Subsequent confocal microscopy revealed that Bm56 was distributed in the outer nuclear membrane and the intranuclear region of infected cells. To investigate the role of Bm56 in virus replication, a Bm56-knockout bacmid of BmNPV was constructed via homologous recombination in Escherichia coli. The Bm56 deletion had no effect on budded virus (BV) production in cultured cells; however, the deletion affected occlusion-body morphogenesis. A larval bioassay demonstrated that the Bm56 deletion did not reduce infectivity, whereas it resulted in a 50 % lethal time that was 16-18 h longer than that of the wild-type bacmid at every dose used in this study. These results indicate that Bm56 facilitates efficient virus production in vivo; however, it is not essential for BV production in vitro.

摘要

家蚕核型多角体病毒ORF56(Bm56)是杆状病毒的核心基因,在所有已完成基因组测序的杆状病毒中高度保守。在感染家蚕核型多角体病毒(BmNPV)的细胞中,感染后12小时(p.i.)即可检测到其转录本,使用抗谷胱甘肽S-转移酶-Bm56融合蛋白的多克隆抗体,在感染后16小时可检测到编码蛋白。蛋白质免疫印迹分析表明,Bm56是多角体衍生病毒核衣壳的结构成分。随后的共聚焦显微镜观察显示,Bm56分布在受感染细胞的外核膜和核内区域。为了研究Bm56在病毒复制中的作用,通过在大肠杆菌中进行同源重组构建了BmNPV的Bm56基因敲除杆粒。Bm56基因缺失对培养细胞中出芽病毒(BV)的产生没有影响;然而,该缺失影响了多角体形态发生。幼虫生物测定表明,Bm56基因缺失并没有降低感染力,而在本研究中使用的每个剂量下,其导致的致死时间比野生型杆粒长16 - 18小时。这些结果表明,Bm56促进体内病毒的高效产生;然而,它对于体外BV的产生并非必需。

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