转基因大豆MON89788检测质粒标准分子构建与应用

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3 700 bp的质粒标准分子,其中重组DNA片段1 029 bp.该质粒标准分子的定性PCR检测灵敏度达到10 copy.结论]该研究构建的质粒标准分子pMD-LM3M5能替代NON89788基体标准品,用于MON89788大豆及其产品的定性PCR检测. Abstract： 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.

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

Genetically modified organisms Plasmid reference molecule MON89788 soybean Event-specific detection