Chen Huiqing, Li Mei, Huang Guoping, Mai Weijun, Chen Keping, Zhou Yajing
Institute of Life Sciences, Jiangsu University, 301# Xuefu Road, Zhenjiang, 212013, China,
Curr Microbiol. 2014 Aug;69(2):158-63. doi: 10.1007/s00284-014-0570-3. Epub 2014 Mar 28.
Orf101 (Bm101) of Bombyx mori nucleopolyhedrovirus (BmNPV) is a highly conserved gene in lepidopteran nucleopolyhedroviruses, but its function remains unknown. In this study, Bm101 was characterized. Transcripts of Bm101 were detected from 24 through 96 h post infection (h p.i.) by RT-PCR. The corresponding protein was also detected from 24 to 96 h p.i. in BmNPV-infected BmN cells by Western blot analysis using a polyclonal antibody against Bm101. Western blot assay of occlusion-derived virus and budded virus (BV) preparations revealed that Bm101 encodes a 28-kDa structural protein that is associated with BV and is located in the envelope fraction of budded virions. In addition, confocal analysis showed that the protein was localized in the cytosol and cytoplasmic membrane in virus-infected cells. In conclusion, the available data suggest that Bm101 is a functional ORF of BmNPV and encodes a protein expressed in the late stage of the infection cycle that is associated with the BV envelope.
家蚕核型多角体病毒（BmNPV）的Orf101（Bm101）是鳞翅目核型多角体病毒中一个高度保守的基因，但其功能尚不清楚。在本研究中，对Bm101进行了特性分析。通过RT-PCR在感染后24至96小时（h p.i.）检测到Bm101的转录本。使用针对Bm101的多克隆抗体，通过蛋白质免疫印迹分析在BmNPV感染的BmN细胞中，在感染后24至96小时也检测到了相应的蛋白质。对多角体衍生病毒和出芽病毒（BV）制剂的蛋白质免疫印迹分析表明，Bm101编码一种28 kDa的结构蛋白，该蛋白与BV相关，位于出芽病毒粒子的包膜部分。此外，共聚焦分析表明，该蛋白定位于病毒感染细胞的细胞质和细胞质膜中。总之，现有数据表明Bm101是BmNPV的一个功能性开放阅读框，编码一种在感染周期后期表达的、与BV包膜相关的蛋白质。
