State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control, MOA and Zhejiang Provincial Key Laboratory of Plant Virology, Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.
Mol Plant Pathol. 2011 Sep;12(7):666-76. doi: 10.1111/j.1364-3703.2010.00699.x. Epub 2011 Jan 31.
The subcellular localization of the 11-kDa protein (p11) encoded by ORF3 of Garlic virus X (GarVX; genus Allexivirus, family Alphaflexiviridae) was examined by confocal microscopy. Granules with intense fluorescence were visible on the endoplasmic reticulum when p11 fused with green or red fluorescent protein (GFP or RFP) was expressed in epidermal cells of Nicotiana benthamiana. Moreover, the p11-RFP granules moved in the cytoplasm, along the cell periphery and through the cell membranes to adjacent cells. A 17-kDa protein (p17) of garlic interacting with p11 was identified by yeast two-hybridization and bimolecular fluorescence complementation assay. When p17 fused to GFP was expressed in epidermal cells of N. benthamiana, it localized to the nucleolus. However, in the presence of GarVX p11, the distribution of p17 changed to that of p11, but did not appear to affect the pattern of movement of p11.
利用共聚焦显微镜技术研究了大蒜病毒 X(GarVX;属 Allexivirus,科 Alphaflexiviridae)ORF3 编码的 11kDa 蛋白(p11)的亚细胞定位。当 p11 与绿色或红色荧光蛋白(GFP 或 RFP)融合表达在本氏烟的表皮细胞中时,内质网上可见到荧光强度很高的颗粒。此外,p11-RFP 颗粒在细胞质中移动,沿着细胞边缘穿过细胞膜进入相邻细胞。通过酵母双杂交和双分子荧光互补实验鉴定了与 p11 相互作用的 17kDa 大蒜蛋白(p17)。当 p17 与 GFP 融合表达在本氏烟的表皮细胞中时,它定位于核仁。然而,在 GarVX p11 的存在下,p17 的分布变为 p11 的分布,但似乎不会影响 p11 的运动模式。