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。
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.
作者
闫伟
龙丽坤
李葱葱
夏蔚
董立明
李飞武
YAN Wei;LONG Li-kun;LI Cong-cong;XIA Wei;DONG Li-ming;LI Fei-wu(Institute of Agricultural Quality Standard and Testing Technology,Jilin Academy of Agricultural Sciences,Changchun 130033,China)
出处
《大豆科学》
CAS
CSCD
北大核心
2019年第5期712-718,共7页
Soybean Science
基金
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)
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
Multitarget real-time PCR
Genetically modified soybean
Rapid analysis
Event-specific identification
