Russell R L, Rohrmann G F
Department of Agricultural Chemistry, Oregon State University, Corvallis 97331.
Virology. 1990 Jan;174(1):177-84. doi: 10.1016/0042-6822(90)90066-z.
A polyclonal antiserum against a trpE fusion protein containing the complete open reading frame of the polyhedron envelope (PE) protein from the nuclear polyhedrosis virus of Orgyia pseudotsugata (OpMNPV) was used for immunogold staining and electron microscopic examination of polyhedra, isolated polyhedron envelopes, and infected insect cells at selected times postinfection. The antiserum specifically stained the peripheral envelope of mature polyhedra and also stained the envelope structure which remained after polyhedra were dissolved in dilute alkaline solutions. In OpMNPV-infected Lymantria dispar cells, the PE protein was detected by 48 hr postinfection (hr p.i.) but specific localization and staining of developing polyhedra were not evident. However, by 72 hr p.i. substantial and preferential staining of the periphery of developing polyhedra was evident even though a distinct polyhedron envelope was not yet observed. In addition, the periphery of fibrillar structures was stained by the PE antiserum. By 96 hr p.i., mature envelopes surrounded polyhedra and these polyhedron envelopes were stained with the PE antibody. The progression of PE protein staining during polyhedron morphogenesis indicates that the PE protein accumulates and becomes associated with developing polyhedra in the nucleus between 48 and 72 hr p.i. Very late in infection the mature polyhedron envelope forms on the polyhedron surface. The apparent affinity of the PE protein for the surface of maturing polyhedra suggests that it may be a major component of the polyhedron envelope or may form the matrix for the deposition of other components which contribute to the mature envelope. Immunogold staining and protease digestion experiments indicate that protein is an essential component of the polyhedron envelope.
针对一种包含来自云杉毒蛾核型多角体病毒（OpMNPV）多面体包膜（PE）蛋白完整开放阅读框的trpE融合蛋白的多克隆抗血清，用于在感染后选定时间对多角体、分离的多面体包膜和受感染昆虫细胞进行免疫金染色和电子显微镜检查。该抗血清特异性地染成熟多角体的外周包膜，也染多角体在稀碱溶液中溶解后残留的包膜结构。在感染OpMNPV的舞毒蛾细胞中，感染后48小时（h p.i.）检测到PE蛋白，但发育中的多角体的特异性定位和染色不明显。然而，到感染后72小时，尽管尚未观察到明显的多面体包膜，但发育中的多角体周边有大量且优先的染色。此外，纤维状结构的周边被PE抗血清染色。到感染后96小时，成熟包膜包围多角体，这些多面体包膜被PE抗体染色。多角体形态发生过程中PE蛋白染色的进展表明，PE蛋白在感染后48至72小时在细胞核中积累并与发育中的多角体相关联。在感染的很晚阶段，成熟的多面体包膜在多面体表面形成。PE蛋白对成熟多面体表面的明显亲和力表明，它可能是多面体包膜的主要成分，或者可能形成其他有助于成熟包膜沉积的成分的基质。免疫金染色和蛋白酶消化实验表明蛋白质是多面体包膜的重要成分。
