Development of an Event-Specific Detection Method for Genetically Modified Maize IE034 by Quantitative Real-Time PCR
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The insect-resistant transgenic maize event IE034 has been proved to be one of the most commercially developed transgenic maize events in China. This study was aimed to develop a stable and reliable quantitative detection method to monitor this new transgenic maize event. Here, we developed a novel event-specific real-time PCR method for this genetically modified maize event IE034. The resulting 134 base pair (bp) amplicon was designed according to the 5′ junction of inserted sequence and flanking maize genome sequence. Standard curve of the IE034 5′ event-specific sequence showed good linear regression and high PCR efficiency when using the IE034 pure line samples as calibrator. The limit of detection (LOD) for the IE034 detection method was estimated at approximately 8 initial template copies, and the limit of quantification (LOQ) was estimated at about 40 copies. The accuracy of this quantitative real-time PCR method was verified by screening four mixed DNA samples with known levels of the IE034 event (5, 1, 0.5, and 0.23 %, respectively). The quantified biases deviated from 8.7 to −12.2 %, and the relative standard deviation (RSD) ranged from 2.7 to 12.7 %. These data indicated that this new-developed IE034 event-specific real-time PCR method is suitable and reliable for the quantification of IE034 maize and its derivates.
KeywordsGenetically modified organism IE034 maize Event-specific method Real-time PCR
Compliance with Ethical Standards
This work was supported by the National Transgenic Plant Special Fund (China) (2014ZX08012-001), Jilin Agricultural Science and Technology Innovation Program (2013). We do appreciate the financial support, and the authors acknowledge the financial support for this study.
Conflict of Interest
Feiwu Li declares that he has no conflict of interest. Likun Long declares that she has no conflict of interest. Wei Yan declares that she has no conflict of interest. Congcong Li declares that he has no conflict of interest. Junfeng Xu declares that he has no conflict of interest. Yunjun Liu declares that he has no conflict of interest. Shihong Zhang declares that he has no conflict of interest. IE034 event-specific sequence had been approved to use by the inventor of the patent (China patent no. CN 102604940 B).
This article does not contain any studies with human participants or animals performed by any of the authors.
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