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[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 research aimed to construct a plasmid standard molecule suitable for the detection of transgenic soybean MON89788. [Method] The 3 ’end specific sequence and 5’ end specific sequence of lectin, MON89788 in soybean were cloned in sequence into pMD18-T vector by qualitative PCR, ligation, transformation and other molecular cloning techniques to obtain plasmid standard molecule pMD-LM3M5 and verify suitability. [Result] 3700bp plasmid standard molecule was obtained, in which the recombinant DNA fragment was 1029bp. This plasmid standard molecule has a qualitative PCR detection sensitivity of 10 copy. [Conclusion] The plasmid standard molecule pMD-LM3M5 constructed in this study could replace MON89788 matrix standard for the qualitative PCR detection of MON89788 soybean and its products.