转OsDREB3基因大豆外源基因拷贝数的确定及标准分子的构建  

Determination of copy numbers of OsDREB3 gene and construction of standard molecular of transgenic OsDREB3 soybean

作　　者：孙喆[1] 刘营[1] 张明辉[1] 李璐[1] 高学军[1] 

机构地区：[1]农业部转基因生物产品成分监督检验测试中心(哈尔滨),东北农业大学农业生物功能基因重点实验室,黑龙江哈尔滨150030

出　　处：《甘肃农业大学学报》2015年第3期61-67,共7页Journal of Gansu Agricultural University

基　　金：转基因生物新品种培育重大专项(2014ZX08004-002-002);农业生物功能基因重点实验室基金开放课题项目(NSGJ2012-08)

摘　　要：为建立抗逆转OsDREB3基因大豆快速稳定的品系特异性检测方法,本试验通过实时Real-time PCR方法,以大豆凝集素基因(lectin)作为内源参照基因,确定了外源基因OsDREB3在转OsDREB3基因大豆基因组中的拷贝数为单拷贝,为建立快速、有效的品系特异性检测方法奠定基础.同时,在原有构建的含4种转基因大豆品系特异性序列的标准分子载体上又增加了转OsDREB3基因大豆品系特异性序列,新构建了含有5种转基因大豆品系特异性序列的标准分子及大豆内参照基因(lectin),已完全满足对现有国内覆盖面最大的5种转基因大豆同时、快速筛查检测工作的需要.To establish a fast and stable detection method for OsDREB3,agenetically modified（GMO）soybean,t the exogenous gene OsDREB3 was identified as a single copy gene in the genome by real-time quantitative PCR method and lectin gene as house-keeping gene,which laid a foundation for a rapid and effective detecting method of event specific genetically modified soybeans.At the same time,OsDREB3 event specific sequences were constructed to the multiple-target plasmid,a standard plasmid with four junction regions of genetically modified soybean events,and thus generate a new standard plasmid which can meet fully the demands of rapid detection work of all the five kinds of genetically modified soybeans.

关 键 词：转OsDREB3基因大豆 外源基因 拷贝数 标准分子 

分 类 号：Q78[生物学—分子生物学]