EXPRESSION AND SUBCELLULAR LOCATION OF THE NSM PROTEIN OF TOMATO SPOTTED WILT VIRUS (TSWV), A PUTATIVE VIRAL MOVEMENT PROTEIN
EXPRESSION AND SUBCELLULAR LOCATION OF THE NSM PROTEIN OF TOMATO SPOTTED WILT VIRUS (TSWV), A PUTATIVE VIRAL MOVEMENT PROTEIN
复制标题
DOI：
10.1006/viro.1994.1162
复制
发表时间：
1994-04-01
期刊：
影响因子：
3.7
通讯作者：
GOLDBACH, R
中科院分区：
文献类型：
--
作者：
KORMELINK, R;STORMS, M;GOLDBACH, R
克隆了番茄斑萎病毒（TSWV）双义M RNA片段上的33.6kDa非结构蛋白（NSM）基因，并在大肠杆菌pET-11 t表达系统中进行了表达。纯化由此产生的蛋白质并用于制备多克隆抗血清。对TSWV感染的Nicotiana rustica植物的Western免疫印迹分析显示，NSM仅在全身感染早期的短时间内合成。虽然NSM被发现与细胞质核衣壳制剂，它是不存在的纯化的病毒颗粒。从年轻的，系统感染的叶片的亚细胞组分的分析表明，NSM的存在下，在组分富集细胞壁和细胞质膜，分别。此外，免疫金标记的组织切片TSWV感染N。rustica植物表明，发现该蛋白与细胞质中的核衣壳聚集体相关，并与胞间连丝密切相关。所获得的数据提供的证据表明，NSM代表TSWV的病毒运动蛋白，参与无包膜核糖核衣壳结构的细胞间运动。(C)1994年出版社出版。
The 33.6-kDa nonstructural (NSM) protein gene, located on the ambisense M RNA segment of tomato spotted wilt virus (TSWV), was cloned and expressed using the Escherichia coli pET-11t expression system. The protein thus produced was purified and used for the production of a polyclonal antiserum. Western immunoblot analyses of TSWV-infected Nicotiana rustica plants showed NSM synthesis only during a short period early in systemic infection. Although NSM was found associated with cytoplasmic nucleocapsid preparations, it was absent from purified virus particles. Analyses of subcellular fractions from young, systemically infected leaves showed the presence of NSM in fractions enriched for cell walls and cytoplasmic membranes, respectively. Furthermore, immunogold labeling of tissue sections of TSWV-infected N. rustica plants showed that this protein was found associated with nucleocapsid aggregates in the cytoplasm and in close association with plasmodesmata. The data obtained provide evidence that NSM represents the viral movement protein of TSWV, involved in cell-to-cell movement of nonenveloped ribonucleocapsid structures. (C) 1994 Academic Press, Inc.