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

转基因大豆MON89788检测质粒标准分子构建与应用(摘要)(英文)

作　　者：李飞武[1] 邵改革[1] 邢珍娟[1] 李葱葱[1] 夏蔚[1] 张明[1] Fei-wu Gai-ge Zhen-juan Cong-cong 

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：《Agricultural Science & Technology》2010年第5期55-58,86,共5页农业科学与技术（英文版）

基　　金：Supported by Major Projects of Cultivating New Varieties by Trans-genic Technology (2008ZX08012-001)~~

摘　　要：[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 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 were 10 copies. [Conclusion] The PRM constructed in this study was suitable for the identification of MON89788 event.[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。

关 键 词：Genetically modified organisms    Plasmid reference molecule    MON89788 soybean    Event-specific detection 

分 类 号：S565.1[农业科学—作物学]