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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 被引量:10
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作者 闫伟 龙丽坤 +3 位作者 李葱葱 夏蔚 董立明 李飞武 《大豆科学》 CAS CSCD 北大核心 2019年第5期712-718,共7页
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。 展开更多
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
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Construction of the Plasmid Reference Molecule for Detection of Transgenic Soybean MON89788 被引量:4
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作者 李飞武 邵改革 +7 位作者 邢珍娟 李葱葱 夏蔚 张明 Fei-wu Gai-ge Zhen-juan Cong-cong 《Agricultural Science & Technology》 CAS 2010年第5期55-58,86,共5页
[Objective] The aim was to construct a plasmid reference molecule (PRM) for detection of transgenic soybean MON89788. [Method] the lectin gene sequence,3'-junction and 5'-junction sequence between host plant D... [Objective] The aim was to construct a plasmid reference molecule (PRM) for detection of transgenic soybean MON89788. [Method] the lectin gene sequence,3'-junction and 5'-junction sequence between host plant DNA integrated DNA of MON89788 soybean were amplified independently,and the three fragments were cloned into the cloning vector pMD18-T in order through molecular manipulation method to construct pMD-LM3M5,the applicability of the constructed novel PRM was tested. [Result] Sequencing confirmation result showed that the PRM was 3 700 bp in length,containing 1 029 bp of recombined DNA fragment. The limits of qualitative detection of the PRM were 10 copies. [Conclusion] The PRM constructed in this study was suitable for the identification of MON89788 event. 展开更多
关键词 Genetically modified organisms Plasmid reference molecule MON89788 soybean event-specific detection
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