Identification of Bombyx mori Bidensovirus VD1-ORF4 Reveals a Novel Protein Associated with Viral Structural Component
Identification of Bombyx mori Bidensovirus VD1-ORF4 Reveals a Novel Protein Associated with Viral Structural Component
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家蚕拜登病毒 VD1-ORF4 的鉴定揭示了一种与病毒结构成分相关的新型蛋白质
DOI：
10.1007/s00284-013-0306-9
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发表时间：
2013-06-01
影响因子：
2.6
通讯作者：
Yao, Qin
中科院分区：
文献类型：
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作者：
Li, Guohui;Hu, Zhaoyang;Yao, Qin
家蚕双核病毒（BmBDV）VD 1-ORF 4（开放阅读框架4，ORF 4）由3，318个核苷酸组成，其编码包含保守的DNA聚合酶基序的预测的1，105个氨基酸的蛋白。然而，其在病毒繁殖中的功能仍然未知。在本研究中，从感染后6至96 h（p.i.）RT-PCR结果显示，5′-RACE显示BmBDV ORF 4的转录起始位点位于起始密码子上游-16个核苷酸处，而3′-RACE显示VD 1-ORF 4的转录终止位点位于终止密码子下游+7个核苷酸处。用制备的抗VD 1-ORF 4氨基酸的抗体对BmBDV感染的家蚕中肠提取物进行Western blot检测，发现3种不同的蛋白质，并在纯化的病毒颗粒中检测到一条分子量约为53 kDa的特异性蛋白带。综上所述，BmBDV VD 1-ORF 4在病毒生命周期中编码三种或更多种蛋白，其中一种是53 kDa蛋白，并被证实是BmBDV病毒粒子的组分。
Bombyx mori bidensovirus (BmBDV) VD1-ORF4 (open reading frame 4, ORF4) consists of 3,318 nucleotides, which codes for a predicted 1,105-amino acid protein containing a conserved DNA polymerase motif. However, its functions in viral propagation remain unknown. In the current study, the transcription of VD1-ORF4 was examined from 6 to 96 h postinfection (p.i.) by RT-PCR, 5′-RACE revealed the transcription initiation site of BmBDV ORF4 to be −16 nucleotides upstream from the start codon, and 3′-RACE revealed the transcription termination site of VD1-ORF4 to be +7 nucleotides downstream from termination codon. Three different proteins were examined in the extracts of BmBDV-infected silkworms midguts by Western blot using raised antibodies against VD1-ORF4 deduced amino acid, and a specific protein band about 53 kDa was further detected in purified virions using the same antibodies. Taken together, BmBDV VD1-ORF4 codes for three or more proteins during the viral life cycle, one of which is a 53 kDa protein and confirmed to be a component of BmBDV virion.