The transgenic glyphosate-tolerant soybean MON87712 event was developed by the agrochemical and agricultural biotechnology company Monsanto (USA) and commercialized in 2013. Due to the absence of matrix-based and genomic DNA-positive reference material for MON87712, it is very difficult to detect and monitor this event. In this study, we developed a recombinant 760-bp linearized plasmid, including 150 bp of the soybean endogenous lectin gene and 610 bp of the exogenous BBX32 gene plus its 3ʹ flanking sequence of MON87712 by In-Fusion cloning technology. In addition, a duplex real-time polymerase chain reaction for the detection of MON87712 and the soybean endogenous lectin gene was established. By using this method, we achieved specific and quantitative detection of MON87712 in 45 other kinds of crops, with a detection limit of 10 copies/μl. This method provides a new technical means for the accurate detection of transgenic soybean MON87712, as well as technical support for the supervision of agricultural transgenic organisms.

中文翻译：

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质粒校准物 pMON87712 对转基因大豆 MON87712 定量检测的适用性


转基因耐草甘膦大豆MON87712事件由农化和农业生物技术公司孟山都（美国）开发并于2013年商业化。由于MON87712缺乏基质和基因组DNA阳性参考材料，检测非常困难并监控该事件。在本研究中，我们通过In-Fusion克隆技术开发了重组的760 bp线性化质粒，包括150 bp的大豆内源凝集素基因和610 bp的外源BBX32基因及其MON87712的3′侧翼序列。此外，还建立了检测MON87712和大豆内源凝集素基因的双重实时聚合酶链反应。利用该方法，我们实现了MON87712在其他45种作物中的特异定量检测，检测限为10拷贝/μl。该方法为转基因大豆MON87712的准确检测提供了新的技术手段，也为农业转基因生物的监管提供了技术支撑。