转基因大豆MON89788检测质粒标准分子的构建与应用  

Construction of the Plasmid Reference Molecule for Detection of Transgenic Soybean MON89788

作　　者：李飞武[1] 邵改革[1] 邢珍娟[1] 李葱葱[1] 夏蔚[1] 张明[1] 

机构地区：[1]吉林省农业科学院,农业部转基因植物环境安全监督检验测试中心,吉林长春130033

出　　处：《安徽农业科学》2010年第23期12330-12333,共4页Journal of Anhui Agricultural Sciences

基　　金：转基因生物新品种培育重大专项(2008ZX08012-001)

摘　　要：[目的]构建适用于转基因大豆MON89788检测的质粒标准分子。[方法]利用定性PCR和连接、转化等分子克隆技术,将大豆内标准基因lectin、MON89788的3′端特异性序列和5′端特异性序列依次克隆到pMD18-T载体上,获得质粒标准分子pMD-LM3M5,并进行适用性验证。[结果]获得了3700bp的质粒标准分子,其中重组DNA片段1029bp。该质粒标准分子的定性PCR检测灵敏度达到10copy。[结论]该研究构建的质粒标准分子pMD-LM3M5能替代MON89788基体标准品,用于MON89788大豆及其产品的定性PCR检测。[Objective] The aim was to construct a plasmid reference molecule （PRM） for detection of transgenic soybean MON89788. [Method]The lectin gene sequence,3＇-junction and 5＇-junction sequence between host plant DNA and integrated DNA of MON89788 soybean were amplified independently,and the three fragments were cloned into the cloning vector pMD18-T in order through molecular manipulation method to construct pMD-LM3M5,the applicability of the constructed novel PRM was tested. [Result] Sequencing confirmation result showed that the PRM was 3 700 bp in length,containing 1 029 bp of recombined DNA fragment. The limits of qualitative detection of the PRM was 10 copy. [Conclusion] The PRM constructed in this study was suitable for the identification of MON89788 event.

关 键 词：转基因生物 质粒标准分子 MON89788大豆 转化体特异性检测 

分 类 号：S565.1[农业科学—作物学]