摘要
将绿色荧光蛋白(GFP)连接于蚕豆萎蔫病毒2号(Broad bean wilt virus2,BBWV-2)VP37蛋白的N-端,构建融合基因GFP-VP37,用农杆菌法在BY-2悬浮细胞内进行表达,激光共聚焦显微镜观察融合蛋白的分布。结果显示:GFP-VP37主要定位于细胞核的周围呈网络状,并在细胞边缘形成点状结构;ER-tracker标记显示VP37在细胞内与内质网共定位;电镜免疫金标记显示VP37蛋白主要定位在细胞质中;用BFA处理转染细胞后,VP37在细胞质及细胞边缘的定位受到抑制。推测内质网参与VP37的细胞内转运和分布。
The green fluorescent protein（GFP） was fused to the N-terminal of Broad bean wilt virus 2 （BBWV-2） VP37 protein. The fusion protein was expressed in the BY-2 suspension cells and the green fluorescence was observed using a confocal laser scanning microscope （CLSM）. In the cells expressing GFP-VP37 protein, GFP was found located along cell membrane and accumulated around the nuclei. ER- Tracker was used to label endoplasmic reticulum （ ER）. The results showed that the VP37 co-localize with ER. Moreover immuno-gold labeling with VP37 antibody indicated that the VP37 locates in the cytoplasm. After treating with brefildin A （BFA） the fusion protein failed to target plamodesmata, Our observation suggests that the ER was involved in BBWV-2 intraceUular transport and distribution.
出处
《电子显微学报》
CAS
CSCD
北大核心
2009年第5期405-409,共5页
Journal of Chinese Electron Microscopy Society
基金
国家自然科学基金资助项目(No.30770091
30270873)~~
