水稻rbcS启动子的克隆及其在转基因水稻中的特异性表达  

Cloning of the promoter of rice rbcS and its specific expression in transgenic rice

作　　者：卢碧霞[1] 张改生[1] 夏勉 马守才[1] 

机构地区：[1]西北农林科技大学农学院,陕西杨凌712100 [2]国家作物分子设计中心,北京100085

出　　处：《西北农林科技大学学报（自然科学版）》2007年第1期96-100,105,共6页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家科技部"863"重大专项(2002AA207004);国家科技部"863"项目(2002AA2C1001)

摘　　要：为将高效特异的启动子用于转基因水稻研究,利用PCR技术从水稻‘中花11’基因组DNA中克隆了rbcS启动子,序列分析表明,扩增片段(2 746 bp)与已报道的该基因序列相应区域的同源性达99.2%。将rbcS启动子与GUS报告基因融合构建了由rbcS启动子引导GUS基因的植物表达载体,经农杆菌介导法导入到水稻中。对转基因水稻植株中GU S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中的特异性表达,其表达水平高于C aMV 35S组成型启动子,而在转基因水稻植株根和种子等器官中不表达或表达活性极弱,表现出明显的组织特异性。To use specific expression of foreign genes of promoters in transgenic rice research,the rice rbcS promoter was isolated from the rice of ZhongHua11 genornic DNA by PCR,and its sequencing indicated that the amplified band （2 746 bp） was 99.20% homologous to the reported ones at the correspondent sequence regions. The cloned rbcS promoter was fused to the 5＇-upstream of GUS （beta-glucuronidase） coding region in a binary vector,and introduced into rice by Agrogacteriurn-mediated transformation. The integration of the rbcS-GUS fusion gene in transgenic rice was confirmed by PCR analysis. The determinations of the GUS activities in the transgenic rice plants indicated rbcS promoter could drive the GUS reporter gene to express in the leaves of transgenic rice plants while exerting no or very weak influence on the expression of the GUS reporter gene, and the expression level of rbcS-GUS fusion gene was significantly stronger than the expression level of CaMV 35S-GUS fusion gene. So rbcS promoter was tissue specific in its expression.

关 键 词：水稻rbcS启动子 转基因水稻 基因表达 GUS活性 

分 类 号：Q74[生物学—分子生物学]