番茄CDT2同源基因RNA干涉载体的构建及遗传转化研究

Construction and transformation of RNAi vector of tomato homologue CDT2

真核生物CDT2是CUL4-DDB1 E3泛素复合体的组成部分,在细胞周期调控、DNA复制与损伤修复中起到重要作用.本研究克隆了番茄CDT2同源基因片段并构建了CDT2基因RNA干涉植物表达载体pBI121-CDT2-RNAi.通过根癌农杆菌介导转入番茄子叶,经组织培养成功获得转基因植株.半定量RT-PCR分析显示,转基因植株叶片内CDT2的表达量明显低于野生型植株.转基因植株叶片叶绿素含量比野生型明显升高.该研究结果揭示番茄CDT2基因的功能做出了新的尝试.

The eukaryote CDT2 gene is part of CUL4-DDB1 E3 ubiquitin complex. They play an important role in cell cycle, DNA replication and repair regulation. In this study, homologue CDT2 gene was cloned in tomato and a RNA interference （RNAi） vector of CDT2 was constructed. The construct was introduced into tomato cv. Ailsa Craig by means of Agrobacterium-mediated transformation. Analysis of semi-quantitative RT-PCR revealed a distinct reduction of endogenous CDT2 expression levels in the RNAi transgenic lines compared to that of wild-type plants. The Chlorophyll content of leaves in the RNAi repression lines was significantly elevated. The results show us a new way to modify fruit tional quality in tomato through gene engineering.