Food Biology Laboratory, Technology Center of Dalian Customs District, Dalian 116001, China.
Key Laboratory of Biotechnology and Bioresources Utilization of Ministry of Education, College of Life Science, Dalian Minzu University, Dalian 116600, China.
Acta Biochim Biophys Sin (Shanghai). 2021 Jul 5;53(7):933-942. doi: 10.1093/abbs/gmab063.
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 3' 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.
抗草甘膦转基因大豆 MON87712 事件由农业化学品和农业生物技术公司孟山都（美国）开发，并于 2013 年商业化。由于缺乏 MON87712 的基质和基因组 DNA 阳性参考物质，因此很难检测和监测该事件。在本研究中，我们通过 In-Fusion 克隆技术开发了一种重组的 760-bp 线性化质粒，包括 150bp 的大豆内源凝集素基因和 610bp 的外源性 BBX32 基因及其 MON87712 的 3'侧翼序列。此外，建立了用于检测 MON87712 和大豆内源凝集素基因的双重实时聚合酶链反应。使用该方法，我们实现了对 45 种其他作物中 MON87712 的特异性和定量检测，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持。
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