[1]闫伟,龙丽坤,李葱葱,等.三种转基因大豆品系的多重荧光PCR检测体系建立[J].大豆科学,2019,38(05):712-718.[doi:10.11861/j.issn.1000-9841.2019.05.0712]
　YAN Wei,LONG Li-kun,LI Cong-cong,et al.Establishment of Detection System for Three Genetically Modified Soybean Linesidentification[J].Soybean Science,2019,38(05):712-718.[doi:10.11861/j.issn.1000-9841.2019.05.0712]
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三种转基因大豆品系的多重荧光PCR检测体系建立

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第38卷 期数: 2019年05期 页码: 712-718 栏目: 出版日期: 2019-09-20
Title:
Establishment of Detection System for Three Genetically Modified Soybean Linesidentification
作者:
（吉林省农业科学院 农业质量标准与检测技术研究所，吉林 长春 130033）
Author(s):
(Institute of Agricultural Quality Standard and Testing Technology, Jilin Academy of Agricultural Sciences, Changchun 130033, China)
关键词:
多重实时荧光PCR; 转基因大豆; 快速检测; 品系特异性鉴定
Keywords:
Multitarget real-time PCR;  Genetically modified soybean;  Rapid analysis;  Event-specific
DOI:
10.11861/j.issn.1000-9841.2019.05.0712
文献标志码:
A
摘要:
转基因大豆MON87701、MON87708、MON87769是我国大豆进口贸易中重点监测的转基因品系，为探讨这3个品系的高效监管技术手段，应用多重实时荧光聚合酶链式反应建立在同一反应体系中同时检测MON87701、MON87708、MON87769的方法。通过分析转基因大豆MON87701、MON87708、MON87769品系的特性序列及大豆内标准基因lectin序列，在此基础上优化设计1组多重PCR引物/探针，并以转基因大豆MON87701、MON87708和MON87769为测试样品，以其它转基因大豆、玉米、水稻、油菜、棉花及非转基因大豆为对照测试样品，经引物/探针浓度优化、特异性、灵敏度等测试，建立多重实时荧光PCR检测体系。结果显示：该多重PCR方法可从各种复杂样品中准确检出相应的靶标转基因大豆成分，对转基因大豆MON87701、MON87708和MON87769的检测灵敏度达到43个DNA拷贝，相当于比0.05% (w/w)。该方法特异性强、灵敏度高，在转基因大豆成分快速检测和品系鉴定中具有很好的应用前景。
Abstract:
Genetically modified soybean lines MON87701, MON87708 and MON87769 are the key monitoring targets in soybean import trade in China. In order to provide a highly efficient technical means for the supervision of these transgenic soybean lines, this study established a multitarget real-time polymerase chain reaction(MT-PCR) assay for simultaneous detection of MON87701, MON87708 and MON87769 in one reaction system. We optimized the primers and probes according to the event-specific border sequences of MON87701, MON87708, MON87769 and the endogenous lectin gene sequence in soybean. The primers/probes concentration optimization, specificity, sensitivity and applicability were tested, respectively. Other transgenic soybean, maize, rice, rapeseed, cotton and non-transgenic soybean were used as test samples in this study. The results showed that the MT-PCR method could accurately detect the corresponding target GM soybean components from various complex samples. The absolute limit of detection for MON87701, MON87708 and MON87769 were 43 copies, equivalent to 0.05% (w/w). This method can be used for the rapid identification of corresponding GM soybean lines.

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备注/Memo


收稿日期：2019-04-04
基金项目：国家转基因生物新品种培育科技重大专项(2018ZX08012-001)；吉林省农业科技创新工程（CXGC2017JQ017）。
第一作者简介：闫伟（1984-），女，硕士，助理研究员，主要从事转基因作物分子检测技术研究。E-mail:jlndyanwei@126.com。通讯作者：李飞武（1982-），男，博士，研究员，主要从事转基因作物安全评价与分子检测研究。E-mail:lifeiwu3394@sina.com。

更新日期/Last Update: 2019-09-20