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Event specific qualitative and quantitative polymerase chain reaction detection of genetically modified MON863 maize based on the 5 '-transgene integration sequence

文献类型： 外文期刊

作者： Yang, LT 1 ; Xu, SC 1 ; Pan, AH 1 ; Yin, CS 2 ; Zhang, KW; Wang, ZY; Zhou, ZG; Zhang, DB;

作者机构： 1.Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, Shanghai 200240, Peoples R China; Nanjing Univ, Dept Biol Sci & Technol, Nanjing 210093, Peoples R China; Shanghai Fisheries Univ, Coll Life Sci, Shanghai 200090, Peoples R China; Shanghai Acad Agr Genet, Key Lab Agr Genet & Breeding, Agro Biotech Res Ctr, Shanghai 201106, Peoples R China; Chinese Acad Agr Sci, Inst Plant Protect, Beijing 100094, Peoples R China

2.Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, Shanghai 200240, Peoples R China; Nanjing Univ, De

关键词： MON863 maize;5 '-transgene integration sequence;event specific;qualitative and quantitative PCR;TAIL-PCR

期刊名称：JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY （ 影响因子：5.279； 五年影响因子：5.269 ）

ISSN： 0021-8561

年卷期： 2005 年 53 卷 24 期

页码：

收录情况： SCI

摘要： Because of the genetically modified organisms (GMOs) labeling policies issued in many countries and areas, polymerase chain reaction (PCR) methods were developed for the execution of GMO labeling policies, such as screening, gene specific, construct specific, and event specific PCR detection methods, which have become a mainstay of GMOs detection. The event specific PCR detection method is the primary trend in GMOs detection because of its high specificity based on the flanking sequence of the exogenous integrant. This genetically modified maize, MON863, contains a Cfy3Bb1 coding sequence that produces a protein with enhanced insecticidal activity against the coleopteran pest, corn rootworm. In this study, the 5'-integration junction sequence between the host plant DNA and the integrated gene construct of the genetically modified maize MON863 was revealed by means of thermal asymmetric interlaced-PCR, and the specific PCR primers and TaqMan probe were designed based upon the revealed 5'-integration junction sequence; the conventional qualitative PCR and quantitative TaqMan real-time PCR detection methods employing these primers and probes were successfully developed. In conventional qualitative PCR assay, the limit of detection (LOD) was 0.1 % for MON863 in 100 ng of maize genomic DNA for one reaction. In the quantitative TaqMan real-time PCR assay, the LOD and the limit of quantification were eight and 80 haploid genome copies, respectively. In addition, three mixed maize samples with known MON863 contents were detected using the established real-time PCR systems, and the ideal results indicated that the established event specific real-time PCR detection systems were reliable, sensitive, and accurate.

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[1]Event-Specific Qualitative and Quantitative Polymerase Chain Reaction Analysis for Genetically Modified Canola T45. Yang, Litao,Pan, Aihu,Zhang, Haibo,Guo, Jinchao,Yin, Changsong,Zhang, Dabing.

[2]Event-specific qualitative and quantitative PCR detection of the GMO carnation (Dianthus caryophyllus) variety Moonlite based upon the 5 '-transgene integration sequence. Li, P.,Jia, J. W.,Jiang, L. X.,Zhu, H.,Bai, L.,Wang, J. B.,Tang, X. M.,Pan, A. H.. 2012

[3]高效、新T-DNA侧翼序列分离技术——Actail-PCR. 杨立勇,刘灶长,周立国,罗华程,罗利军. 2009

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