Pearson M N, Russell R L, Rohrmann G F, Beaudreau G S
Department of Agricultural Chemistry, Oregon State University, Corvallis 97331.
Virology. 1988 Dec;167(2):407-13.
A series of monoclonal antibodies was produced against proteins from polyhedra-derived virions of the multicapsid nuclear polyhedrosis virus of Orgyia pseudotsugata (OpMNPV). Two of these antibodies (214 and 236) reacted with a protein of 39 kDa on Western blots of electrophoretically separated OpMNPV viron proteins derived from both budded and polyhedra-derived virions. This protein appears to be a major component of both BV and PDV. One of the p39 antibodies was used to characterize p39 synthesis in infected Lymantria dispar cells by using Western blots and immunofluorescent staining. The p39 protein was detected by immunofluorescence microscopy at 24 hr postinfection. By 48 hr, p39 was detected primarily in cell nuclei with little or no detectable staining of the cytoplasm. The two MAbs were used to identify three immunoreactive clones from a lambda gt11 expression library of OpMNPV DNA. By hybridization of insert DNA from three lambda gt11 clones to blots of restriction digests of OpMNPV genomic DNA, the location of the 39-kDa gene was mapped on the OpMNPV genome. Using the lambda gt11 insert DNAs and the monoclonal antibodies, the p39 genes and proteins of OpMNPV and Autographa californica NPV (AcMNPV) were shown to be closely related in size, DNA sequence, and antigenicity. One of the p39 monoclonal antibodies cross-reacted with a host cell protein associated with the condensed chromosomes present during mitosis.
制备了一系列针对云杉毒蛾多核衣壳核型多角体病毒（OpMNPV）多角体衍生病毒粒子蛋白的单克隆抗体。其中两种抗体（214和236）在源自出芽病毒粒子和多角体衍生病毒粒子的电泳分离OpMNPV病毒粒子蛋白的Western印迹上，与一种39 kDa的蛋白发生反应。这种蛋白似乎是芽生病毒粒子（BV）和多角体衍生病毒粒子（PDV）的主要成分。利用Western印迹和免疫荧光染色，其中一种p39抗体被用于表征感染舞毒蛾细胞中p39的合成情况。感染后24小时通过免疫荧光显微镜检测到p39蛋白。到48小时时，p39主要在细胞核中被检测到，细胞质中几乎没有或没有可检测到的染色。这两种单克隆抗体被用于从OpMNPV DNA的λgt11表达文库中鉴定出三个免疫反应性克隆。通过将来自三个λgt11克隆的插入DNA与OpMNPV基因组DNA的限制性消化印迹杂交，将39 kDa基因的位置定位在OpMNPV基因组上。利用λgt11插入DNA和单克隆抗体，显示OpMNPV和苜蓿银纹夜蛾核型多角体病毒（AcMNPV）的p39基因和蛋白在大小、DNA序列和抗原性方面密切相关。其中一种p39单克隆抗体与一种与有丝分裂期间存在的浓缩染色体相关的宿主细胞蛋白发生交叉反应。
