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

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.