芸薹属类黄酮3′-羟化酶基因家族RNA干扰载体的构建  

Construction of RNAi Vector of Brassica Flavonoid 3'-Hydroxylase Gene Family

作　　者：陈倩[1] 闫楠[1] 马丽娟[1] 柴友荣[1] 

机构地区：[1]西南大学农学与生物科技学院,农业部生物技术与作物品质改良重点实验室,重庆市作物品质改良重点实验室,重庆市油菜工程技术研究中心,重庆400716

出　　处：《生物技术》2010年第3期8-11,共4页Biotechnology

基　　金：国家高技术研究发展计划(863计划)专题课题("甘蓝型油菜类黄酮途径功能基因克隆及黄籽性状代谢工程研究";2006AA10Z110);重庆市科技攻关项目("油菜分子育种及品质检测技术研究";CSTC2009AB1030)资助

摘　　要：目的:构建芸薹属类黄酮3′羟化酶(F3′H)基因家族的RNAi载体。方法:采用PCR克隆了607bp的芸薹属F3’H基因家族的RNAi片段,将其反义片段、正义片段分别采用NcoⅠ+AatⅡ、BamHⅠ+XbaⅠ插入到改进型植物RNAi基础载体pF-GC5941M的启动子与间隔区、间隔区与终止子之间,形成11366bp的重组载体pFGC5941M-BF3′HI(简称为pBF3′HI),复合PCR鉴定后转化根癌农杆菌,获得工程菌株。结果:载体构建成功。结论:构建了RNAi载体pBF3′HI,可用于芸薹属植物花色、种皮色泽、茎叶色彩等性状的机理研究和遗传修饰。Objective：Constructing of an RNAi vector of Brassica flavonoid 3＇-Hydroxylase（F3＇H） gene family.Method：An 607bp RNAi fragment targeted to Brassica F3＇H gene family was cloned.Its antisense and sense fragments were inserted to the promoter-spacer and spacer-terminator cloning sites of an improved plant RNAi basic vector pFGC5941M using NcoⅠ ＋ AatⅡ and BamHⅠ ＋ XbaⅠ double digestions respectively.The resulted 11 366 bp recombinant vector was pFGC5941M-BF3＇HⅠ（simplified as pBF3＇HI）.After verification by multiple PCR,it was transformed into Agrobacterium to form engineering strain.Result：The vector was successfully constructed.Conclusion：The RNAi pBF3＇HI was constructed,which can be used for mechanism dissection and genetic modification of Brassica flower colors,seed coat pigments and plant surface anthocyanin pigmentation.

关 键 词：芸薹属 类黄酮3′-羟化酶(F3′H) RNAI 载体 

分 类 号：Q782[生物学—分子生物学]