转基因抗草甘膦大豆的特定事件实时检测与定量分析

Event-specific real-time detection and quantification of genetically modified Roundup Ready soybean.

作者信息

Huang Chia-Chia, Pan Tzu-Ming

机构信息

Institute of Microbiology and Biochemistry, National Taiwan University, Taipei, Taiwan 106.

出版信息

J Agric Food Chem. 2005 May 18;53(10):3833-9. doi: 10.1021/jf048580x.

DOI:10.1021/jf048580x

PMID:15884804

Abstract

The event-specific real-time detection and quantification of Roundup Ready soybean (RRS) using an ABI PRISM 7700 sequence detection system with light upon extension (LUX) primer was developed in this study. The event-specific primers were designed, targeting the junction of the RRS 5' integration site and the endogenous gene lectin1. Then, a standard reference plasmid was constructed that carried both of the targeted sequences for quantitative analysis. The detection limit of the LUX real-time PCR system was 0.05 ng of 100% RRS genomic DNA, which was equal to 20.5 copies. The range of quantification was from 0.1 to 100%. The sensitivity and range of quantification successfully met the requirement of the labeling rules in the European Union and Taiwan.

摘要

本研究开发了一种利用带有延伸发光（LUX）引物的ABI PRISM 7700序列检测系统对抗草甘膦转基因大豆（RRS）进行事件特异性实时检测和定量分析的方法。设计了事件特异性引物，靶向RRS 5'整合位点与内源性基因凝集素1的连接处。然后构建了携带两个靶向序列的标准参考质粒用于定量分析。LUX实时荧光定量PCR系统的检测限为0.05 ng的100% RRS基因组DNA，相当于20.5个拷贝。定量范围为0.1%至100%。灵敏度和定量范围成功满足了欧盟和台湾地区标签规定的要求。