Russell R L, Rohrmann G F
Department of Agricultural Chemistry, Oregon State University, Corvallis 97331-7301, USA.
Virology. 1997 Jun 23;233(1):210-23. doi: 10.1006/viro.1997.8599.
Polyclonal antiserum produced against preoccluded virions from the Orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (OpMNPV) was used to screen an OpMNPV lambda gt11 expression library. The insert from one of the immunoreactive phage isolates hybridized to OpMNPV orf86 (p91), a 2460-bp (819 amino acids) open reading frame that encodes a predicted protein of 91 kDa. Antibodies generated against a maltose binding protein-P91 fusion detected a band of approximately 91 kDa on Western blots of extracts of OpMNPV-infected Lymantria dispar cells. This band was first observed at 18 hr p.i. and was present at all later time points. Similar results using this antiserum were seen with a time course of Autographa californica-infected Spodoptera frugiperda cells. Localization of P91 by confocal immunofluorescence microscopy showed that the protein was concentrated near the nuclear membrane and at late times p.i. was most concentrated near polyhedra. Immunoelectron microscopy indicated that P91 was present in both the capsid and envelope surrounding the capsid of occlusion-derived virions. Fractionation studies employing NP-40 and Western blot analysis indicated that P91 was associated with the capsid structure.
用针对云杉芽枯叶蛾多核衣壳核型多角体病毒（OpMNPV）的预封闭病毒粒子产生的多克隆抗血清筛选OpMNPV λgt11表达文库。从一种免疫反应性噬菌体分离物中获得的插入片段与OpMNPV orf86（p91）杂交，orf86是一个2460 bp（819个氨基酸）的开放阅读框，编码一种预测分子量为91 kDa的蛋白质。针对麦芽糖结合蛋白-P91融合体产生的抗体在感染OpMNPV的舞毒蛾细胞提取物的蛋白质印迹上检测到一条约91 kDa的条带。该条带在感染后18小时首次出现，并在所有后续时间点都存在。用这种抗血清对感染苜蓿银纹夜蛾核型多角体病毒的草地贪夜蛾细胞进行时间进程实验也得到了类似结果。通过共聚焦免疫荧光显微镜对P91进行定位显示，该蛋白集中在核膜附近，在感染后期，在多角体附近最为集中。免疫电子显微镜表明，P91存在于包涵体衍生病毒粒子的衣壳以及围绕衣壳的包膜中。采用NP-40的分级分离研究和蛋白质印迹分析表明，P91与衣壳结构相关。
