Han Yingying, Xia Hengchuan, Tang Qi, Lü Peng, Ma Shangshang, Yang Yanhua, Shao Dandan, Ma Quanbing, Chen Keping
Institute of Life Sciences, Jiangsu University, 301 Xuefu Road, Zhenjiang, 212013, China,
Curr Microbiol. 2014 Jul;69(1):56-62. doi: 10.1007/s00284-014-0551-6. Epub 2014 Mar 5.
The Bm111 of Bombyx mori nucleopolyhedrovirus (BmNPV) encodes a small polypeptide (70 amino acids) of which the function remains unknown. To characterize its function, multiple sequence alignments were performed, and the predicted protein was found to share amazingly high (98 %) sequence identity with the Bombyx mandarina nucleopolyhedrovirus ORF110 (Boma110) but negligible with proteins of other insect viruses, indicating the close relationship between these two NPVs with silkworm larvae. The transcription of Bm111 was detected as early as 3 hpi in BmNPV-infected BmN cells, suggesting it is an early gene. To investigate the role of Bm111 in baculovirus life cycle, a Bm111-knockout virus was constructed by bacmid recombination in Escherichia coli. The results showed that knockout of the Bm111 did not affect the replication of virus DNA, but significantly extended the death time of infected silkworm larvae compared to the wild-type or rescued viruses. We also successfully expressed the recombinant protein Bm111 in E. coli to provide sufficient material for subsequent studies. Taken together, our data indicate that Bm111 only affects the virulence of BmNPV, but not its replication.
家蚕核型多角体病毒(BmNPV)的Bm111编码一种小多肽(70个氨基酸),其功能尚不清楚。为了表征其功能,进行了多序列比对,发现预测的蛋白质与野桑蚕核型多角体病毒ORF110(Boma110)具有惊人的高序列同一性(98%),但与其他昆虫病毒的蛋白质同一性可忽略不计,这表明这两种NPV与家蚕幼虫之间关系密切。在BmNPV感染的BmN细胞中,早在感染后3小时就检测到了Bm111的转录,表明它是一个早期基因。为了研究Bm111在杆状病毒生命周期中的作用,通过在大肠杆菌中进行杆粒重组构建了一种Bm111敲除病毒。结果表明,敲除Bm111不影响病毒DNA的复制,但与野生型或拯救病毒相比,显著延长了感染家蚕幼虫的死亡时间。我们还成功地在大肠杆菌中表达了重组蛋白Bm111,为后续研究提供了充足的材料。综上所述,我们的数据表明Bm111仅影响BmNPV的毒力,而不影响其复制。