Shirasawa-Seo Naomi, Sano Yoshitaka, Nakamura Shigeo, Murakami Taka, Seo Shigemi, Ohashi Yuko, Hashimoto Yoshifumi, Matsumoto Tsuguo
Miyagi Prefectural Agriculture and Horticulture Research Center, Takadate-kawakami, Natori, Miyagi 981-1243, Japan.
Laboratory of Plant Pathology, Faculty of Agriculture, Niigata University, Ikarashi, Niigata 950-2181, Japan.
J Gen Virol. 2005 Jun;86(Pt 6):1851-1860. doi: 10.1099/vir.0.80790-0.
Predicted promoter regions of Milk vetch dwarf virus (MDV) components (C1-C11) were isolated and fused with a beta-glucuronidase (GUS) reporter gene and the characteristics of the promoters were examined. In transgenic tobacco calli, promoters of MDV C4 (encoding a cell-cycle link protein), C5 and C7 (both encoding unknown proteins), C6 (encoding a nuclear-shuttle protein) and C8 (encoding a movement protein) generated a stronger level of GUS expression than the Cauliflower mosaic virus 35S RNA promoter (P35S). In leaves of transgenic tobacco plants, the promoters of C5 and C8 conferred a level of GUS activity comparable to that of P35S. Histochemical GUS analysis showed that the promoters of C4-C9, the latter encoding a capsid protein, were active in phloem and meristematic tissue. The promoter of C8 was also active in mesophyll and cortex cell types. A low level of activity was found for the promoters of C11, which encodes a master replication-initiator protein (Rep), and C1, C2, C3 and C10, which encode additional Reps, in both transgenic tobacco calli and plants.
分离出紫云英矮缩病毒(MDV)组分(C1 - C11)的预测启动子区域,并将其与β - 葡萄糖醛酸酶(GUS)报告基因融合,然后检测这些启动子的特性。在转基因烟草愈伤组织中,MDV C4(编码一种细胞周期连接蛋白)、C5和C7(均编码未知蛋白)、C6(编码一种核穿梭蛋白)和C8(编码一种运动蛋白)的启动子产生的GUS表达水平高于花椰菜花叶病毒35S RNA启动子(P35S)。在转基因烟草植株的叶片中,C5和C8的启动子赋予的GUS活性水平与P35S相当。组织化学GUS分析表明,C4 - C9的启动子在韧皮部和分生组织中具有活性,C9编码一种衣壳蛋白。C8的启动子在叶肉和皮层细胞类型中也具有活性。在转基因烟草愈伤组织和植株中,编码主要复制起始蛋白(Rep)的C11以及编码其他Reps的C1、C2、C3和C10的启动子活性较低。