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Event-Specific Qualitative and Quantitative Detection of Genetically Modified Rice G6H1

Xu, Xiaoli, Chen, Xiaoyun, Lai, Yongmin, Zhang, Qing, Wu, Yuhua, Li, Jun, Peng, Cheng, Wang, Xiaofu, Miao, Qingmei, Liu, Hui, Xu, Junfeng
Food analytical methods 2019 v.12 no.2 pp. 440-447
3-phosphoshikimate 1-carboxyvinyltransferase, DNA, analytical methods, detection limit, genes, haploidy, host plants, monitoring, quantitative polymerase chain reaction, rice, transgenic plants, China
The genetically modified rice G6H1 expressing a fused protein of Cry1Ab/Vip3H and a 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) G6 is a genetically modified event that has been approved for preproduction field testing in China. The purpose of this study was to establish an event-specific qualitative and quantitative detection method that could provide a stable, reliable system for monitoring this new transgenic event. In this study, event-specific qualitative and quantitative detection methods based on the 3′ integration junction sequence between host plant DNA and the integrated gene were developed. The limit of detection (LOD) of qualitative PCR was assessed to be 0.1%. The LOD of quantitative PCR was estimated to be ten haploid genome copies. The quantitative PCR detection method was verified with three mixed rice samples with known G6H1 contents, and the results agreed with the expected values. Evaluation of specificity and sensitivity indicated that the developed qualitative and quantitative PCR methods are reliable and can be used for the detection and quantification of G6H1.