番茄microRNA基因沉默载体的构建方法  

A MicroRNA-Based Gene Silencing Vector Construction Method in Tomato

作　　者：王云香[1] 左进华[1] 鞠正[1] 高超[1] 朱毅[1] 朱本忠[1] 

机构地区：[1]中国农业大学食品科学与营养工程学院,北京100083

出　　处：《北方园艺》2012年第3期101-104,共4页Northern Horticulture

基　　金：国家自然科学基金面上资助项目(30972037)

摘　　要：该试验提出了一种简单且快速构建番茄中人工miRNA表达载体的方法,以包含miR319a前体骨架的质粒pRS300为模板,以重叠PCR技术来扩增miR1917的前体。miR1917的靶基因是LeCTR1,是乙烯反应中的一种负调控因子。含有miR1917的重组前体成功地转化到表达载体pGreen0029-35s中,通过植物根癌农杆菌C58介导的叶盘法成功获得了转基因植株。此种方法可为番茄中miRNA的作用机理研究提供一定的理论基础。A microRNA-baesd method for easy and rapid construction of tomato miRNA vector was described.The plasmid PRS300 which contained the miR319 aprecursor was used as the temple,the overlapping PCR amplification for the miR1917 whose target gene was LeCTR1 which were key negative regulators of ethylene responses was processed,and the reconstructed precursor was successfully transformed to the expression vector pGreen0029-35s.The transgenic plants were successfully obtained by the leaf disc method mediated by Agrobacterium tumefaciens C58.This method could provide a theoretical basis for the miRNA mechanisms of action research in tomato.

关 键 词：人工microRNA 基因沉默 番茄 载体构建 

分 类 号：S641.203.6[农业科学—蔬菜学]