Development of an Event-Specific Detection Method for Genetically Modified Maize IE034 by Quantitative Real-Time PCR
文献类型： 外文期刊
作者：
Li, Feiwu
1
;
Long, Likun
2
;
Yan, Wei
2
;
Li, Congcong
2
;
Xu, Junfeng
3
;
Liu, Yunjun
4
;
Zhang, Shihong
1
;
作者机构： 1.Jilin Univ, Coll Plant Sci, Changchun 130062, Peoples R China
2.Jilin Acad Agr Sci, Inst Agr Qual Stand & Testing Technol, Changchun 130033, Peoples R China
3.Zhejiang Acad Agr Sci, Inst Qual Stand Agr Prod, Hangzhou 310021, Zhejiang, Peoples R China
4.Chinese Acad Agr Sci, Inst Crop Sci, Beijing 100081, Peoples R China
关键词： Genetically modified organism;IE034 maize;Event-specific method;Real-time PCR
期刊名称：FOOD ANALYTICAL METHODS （ 影响因子：3.366； 五年影响因子：3.07 ）
ISSN：
年卷期：
页码：
收录情况： SCI
摘要： 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.
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