山东农业大学学新新士学位论文.pdfVIP

山东农业大学学士学位论文 水稻 RS6 基因反义表达载体的构建及遗传转化 1999级生物技术专业 苏英华 指 导 教 师: 张宪省 教授 李兴国 副教授 摘要： 本实验旨在利用反义 RNA 技术对水稻种子特异基因 RS6 进行功能分析。首先以 RT-PCR 技 术克隆到 RS6 特异片段，然后将此特异片段反向插入表达载体 Ubiquitin-1301 中，构建起RS6 反义 表达载体 Ubiquitin-1301-RS6 。利用农杆菌介导的方法转化水稻（Oryza sativa L. ）的胚性愈伤组织。 通过抗性筛选，以期获得转基因植株，并通过对转基因植株的表型分析，确定 RS6 的功能。目前已 获得部分抗性愈伤组织。 关键词：水稻；反义RNA ；表达载体；遗传转化 Construction of expression vector with anti-RS6 and Agrobacterium-mediated transformation of rice (Oryza sativa L.) Biotechnology, 1999 Ying hua Su Directors: Xian sheng Zhang professor Xing guo Li adjunct professor Abstract: The function of RS6, a gene expressed preferentially in rice （Oryza sativa L. ）seed was characterized by antisense RNA technology. 3’ UTR of RS6 gene was cloned with RT-PCR and the antisense sequence was inserted into the Ubiquitin-1301 expression vector. The vector with antisense specific fragment of RS6 was constructed and named Ubiquitin-RS6. The antisense construct of RS6 was then introduced into rice (Oryza sativa cv. Zhong Hua 11) calli by means of an Agrobacterium -mediated transformation. According to the phenotype of transgenic plants, we can characterize its biological function. So far, we have got the transgenic calli successfully. Key words: Oryza sativa L.; antisense RNA; expression vector; transformation 1 山东农业大学学士学位论文 1. 引言 水稻是基因组较小的单子叶植物，也是第一大粮食作物，且已建立成熟的遗传转化系统，是研 究禾本科作物以及单子叶植物发育、遗传和分子生物学的模式植物。目前，水稻基因