水稻rbcS启动子控制的外源基因在转基因水稻中的特异性表达  

Specific Expression of the Foreign Gene Regulated by the Rice rbcS Promoter in Transgenic Rice

作　　者：刘巧泉[1] 于恒秀[1] 张文娟[2] 王红梅[2] 顾铭洪[1] 

机构地区：[1]扬州大学江苏省植物功能基因组学重点实验室,生物科学与技术学院扬州225009 [2]扬州大学生物科学与技术学院,扬州225009

出　　处：《植物生理与分子生物学学报》2005年第3期247-253,共7页Journal Of Plant Physiology and Molecular Biology

基　　金：国家"863"计划(No.2004AA212092);国家自然科学基金(Nos.30470992;30300226);江苏省自然科学基金(Nos.BG2002301;BK2003214);教育部霍英东教育基金(94019)资助。~~

摘　　要：为将不同启动子用于转基因水稻的研究,从武运粳8号水稻中克隆了Rubisco小亚基基因(rbcS)的5'上游调控区,构建了由rbcS启动子引导的GUS融合基因,并经农杆菌介导导入到水稻中。对转基因水稻植株中GUS活性的定性与定量测定结果表明,rbcS启动子可驱动GUS报告基因在转基因水稻植株叶片和叶鞘内的叶肉细胞中特异性高效表达,而在茎、根和种子等器官中不表达或表达活性极弱,表现出明显的组织与细胞特异性。结果还表明,光诱导处理可明显提高rbcS启动子启动的外源基因的表达量。To use different types of promoters intransgenic rice research, the 5'-upstream regula-tion region of rice Rubisco small subunit gene(rbcS) was cloned from a Chinese cultivarWuyunjing 8, and its sequences were confirmedby comparison with the known genome se-quences of both japonica and indica rice. Thecloned rbcS promoter was fused to the 5'-up-stream of GUS (beta-glucuronidase) coding re-gion in a binary vector (Fig.1), and introducedinto rice by Agrogacterium-mediatedtransformation. The integration of the rbcS-GUSfusion gene in transgenic rice was confirmed byPCR analysis (Fig.2). The results of both his-tochemical staining and quantitative analysis ofGUS activity showed that the expression levelof GUS fusion gene was significantly strongerin leaf blade and sheath than in other organs oftransgenic rice plants, and the GUS activity wasrestricted to the mesophyll cells of leaf tissue(Figs.3, 4), which showed that the rice rbcS pro- moter could control not only the tissue- but alsothe cell-specific expression of foreign genes intransgenic rice. The present results also demon-strated that light induction had a significant ef-fect on the enhancement of transgene’s expres-sion when regulated by the rice rbcS promoterin transgenic rice (Fig.5). Our results showed thatthe rice rbcS promoter might be very useful forthe expression of target genes in transgenic rice,with particularly high efficiency in leaf tissues.

关 键 词：水稻rbcS启动子 转基因水稻 基因表达 GUS活性 光诱导 

分 类 号：S511[农业科学—作物学]