Kormelink R, Kitajima E W, De Haan P, Zuidema D, Peters D, Goldbach R
Department of Virology, PD Wageningen, The Netherlands.
Virology. 1991 Apr;181(2):459-68. doi: 10.1016/0042-6822(91)90878-f.
The open reading frame located in the viral strand of the ambisense S RNA of tomato spotted wilt virus (TSWV), was cloned into transfer vector pAc33DZ1 and inserted downstream of the polyhedrin promoter in the Autographa californica nuclear polyhedrosis virus genome. Recombinant baculoviruses were obtained that showed a high-level expression of a 52.4-kDa protein corresponding to the inserted TSWV gene. The viral protein thus produced was purified and injected into rabbits to raise antibodies. Western immunoblot analyses of extracts from TSWV-infected plants demonstrated that the 52.4-kDa TSWV-specific polypeptide represents a nonstructural protein (denoted NSs), being absent in purified virus particles. Immunogold labeling of tissue sections of TSWV-infected Nicotiana rustica plants showed that this protein was, depending on the virus isolate, either found dispersed throughout the cytoplasm or associated with fibers which appeared as elongated flexible filaments or paracrystalline rods.
番茄斑萎病毒(TSWV)双义S RNA病毒链上的开放阅读框被克隆到转移载体pAc33DZ1中,并插入到苜蓿银纹夜蛾核型多角体病毒基因组的多角体蛋白启动子下游。获得了重组杆状病毒,其显示出对应于插入的TSWV基因的52.4 kDa蛋白的高水平表达。如此产生的病毒蛋白被纯化并注射到兔子体内以产生抗体。对TSWV感染植物提取物的Western免疫印迹分析表明,52.4 kDa的TSWV特异性多肽代表一种非结构蛋白(称为NSs),在纯化的病毒颗粒中不存在。对TSWV感染的黄花烟草植物组织切片的免疫金标记显示,根据病毒分离株的不同,这种蛋白要么分散在整个细胞质中,要么与呈现为细长的柔性细丝或类晶体棒状的纤维相关联。