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转OsDREB3基因大豆外源基因拷贝数的确定及标准分子的构建 预览

Determination of copy numbers of OsDREB3 gene and construction of standard molecular of transgenic OsDREB3 soybean
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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.
作者 孙喆 刘营 张明辉 李璐 高学军 SUN Zhe;LIU Ying;ZHANG Ming-hui;LI Lu;GAO Xue-jun(Superivision and Test Center (Harbin) for Molecular Characteristics of Genetically Modified Plants, Ministry of Agriculture, Key Laboratory of Agricultural Genomics, Northeast Agricultural University, Harbin 150030, China)
出处 《甘肃农业大学学报》 CAS CSCD 北大核心 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)
关键词 转OsDREB3基因大豆 外源基因 拷贝数 标准分子 OsDREB3 transgenic soybean exogenous gene copy number standard molecular
作者简介 孙喆(1980-),女,实验师,硕士,从事转基因检测技术方面的研究.E-mail:sunzhel998@163.com 通信作者:刘营,女,助理研究员,博士,从事转基因检测技术方面的研究.E-mail:lyneau@126.com
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