Gong Yingxue, Li Zhaofei, Wang Lihua, Pan Lijing, Yang Kai, Pang Yi
State Key Laboratory for Biocontrol, Zhongshan University, Guangzhou 510275, P.R.China.
Virus Genes. 2003 Oct;27(2):115-23. doi: 10.1023/a:1025773324560.
Spodoptera litura multicapsid nucleopolyhedrovirus (SpltMNPV) ORF125 (designated as the bro-b gene) is a member of the unique multigene family called the baculovirus repeated ORFs (bro) family. Computer-assisted analysis revealed that BRO-B contains a conserved Bro-N domain in its N-terminus and a single-stranded DNA (ssDNA) binding motif in the middle region. In vitro the bro-b gene transcription was present at 12 h post-infection (p.i.) and remained detectable up to 96 h p.i. Western blot analysis of BRO-B expression with an antiserum made against 6xHis tagged BRO-B expressed in Escherichia coli showed that it was present from 12 h through 96 h p.i. in vitro. Structural localization revealed that BRO-B could be found in the nucleocapsid components of both occlusion-derived virus (ODV) and budded virus (BV). Immunofluorescence analysis indicated that BRO-B is located only in the nucleus of infected S. litura cells. Furthermore, Western blot analysis indicated that BRO-B was associated with nuclear structures. These results suggested that BRO-B might be a nuclear-associated protein.
斜纹夜蛾多粒包埋核多角体病毒（SpltMNPV）的ORF125（命名为bro - b基因）是杆状病毒重复开放阅读框（bro）家族这一独特多基因家族的成员。计算机辅助分析表明，BRO - B在其N端含有一个保守的Bro - N结构域，在中间区域含有一个单链DNA（ssDNA）结合基序。在体外，bro - b基因转录在感染后12小时（p.i.）出现，并在感染后96小时内均可检测到。用针对在大肠杆菌中表达的6xHis标记的BRO - B制备的抗血清对BRO - B表达进行的蛋白质免疫印迹分析表明，在体外感染后12小时至96小时内均有该蛋白存在。结构定位显示，BRO - B可在包涵体衍生病毒（ODV）和出芽病毒（BV）的核衣壳成分中找到。免疫荧光分析表明，BRO - B仅位于受感染的斜纹夜蛾细胞的细胞核中。此外，蛋白质免疫印迹分析表明，BRO - B与核结构相关。这些结果表明，BRO - B可能是一种核相关蛋白。
