大豆fad3c基因沉默载体的构建  

Construction of fad3c Gene Silencing Vector in Soybean

作　　者：杜景红[1,2] 刘丽君[1] 

机构地区：[1]黑龙江省农业科学院博士后科研工作站,黑龙江哈尔滨150080 [2]黑龙江大学农业资源与环境学院,黑龙江哈尔滨150080

出　　处：《大豆科学》2013年第1期28-32,共5页Soybean Science

基　　金：黑龙江省农业科学院博士后科研启动经费

摘　　要：构建大豆fad3c基因的沉默载体,旨在为培育生物安全的高油酸低亚麻酸转基因大豆奠定基础。从高蛋白大豆黑农35中克隆大豆种子特异表达启动子GY1;从高油大豆黑农37中克隆fad2-1b基因内含子1;从5个转基因受体大豆中克隆fad3c基因片段,作为正向臂和反向臂。构建筛选标记基因为bar,启动子为GY1,内含子为fad2-1b基因内含子1的双T-DNA载体。将双T-DNA载体转化大肠杆菌DH5α,酶切和序列分析鉴定表明扩增得到的重组质粒正确,命名为pDT-GFAD3I。Soybean fad3c gene silencing vector was constructed for breeding high oleic acid and low linolenic transgenic soy- beans with biosafety. Seed-specific promoter GY1 of soybean was amplified from high protein soybean ＇ Heinong 35 ＇ ;fad2-1 b gene intron 1 was amplified from high oil soybean ＇ Heinong 37＇ ;fad3c gene fragment was amplified from five transgenic ac- ceptors of soybean,which was used as forward arm and reverse arms. These target fragments were cloned into the TA cloning vector with selection marker bar gene, promoter GY1 and fad2-1b gene intron 1. Then the recombination plasmid was intro- duced into E. coli DHSa, and the plasmid was verified by enzymes digestion and sequence analysis. The recombination plasmid was named pDT-GFAD3I.

关 键 词：大豆fad2-1b fad3c RNAI 载体构建 

分 类 号：S565.1[农业科学—作物学]