转基因玉米2A-5特异性PCR方法的建立

doi:10.19586/j.2095-2341.2020.0067

生物技术进展 ›› 2020, Vol. 10 ›› Issue (4): 363-370.DOI: 10.19586/j.2095-2341.2020.0067

转基因玉米2A-5特异性PCR方法的建立

武文艳^§,刘新香^§,周秒依^*,刘金香,刘亚

北京市农林科学院玉米研究中心, 玉米DNA指纹及分子育种北京市重点实验室, 北京 100097

收稿日期:2020-05-09 出版日期:2020-07-25 发布日期:2020-06-11
通讯作者: 周秒依 E-mail: monazhou001@126.com
作者简介:§武文艳与刘新香为本文共同第一作者。武文艳 E-mail: wenyansunshine@163.com；刘新香 E-mail: maizelxx@126.com
基金资助:
国家转基因生物新品种培育重大专项（2016ZX08003001）。

Establishment of Event-specific PCR Detection Method of Transgenic Maize Line 2A-5

WU Wenyan, LIU Xinxiang, ZHOU Miaoyi, LIU Jinxiang, LIU Ya

Beijing Key Laboratory of Maize DNA Fingerprinting and Molecular Breeding, Maize Research Center, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China

Received:2020-05-09 Online:2020-07-25 Published:2020-06-11

摘要/Abstract

摘要： 根据转基因玉米2A-5的旁侧序列信息，设计并筛选出最佳特异性引物及Taqman探针组合2A-5-5-QF8、2A-5-5-QR8、2A-5-5-QP8，优化了该引物探针组合的反应体系，建立了转基因玉米2A-5转化体特异性PCR检测方法。将特异性引物及Taqman探针组合用于qRT-PCR和ddPCR技术,研究了转基因玉米2A-5转化体的定量检测方法，发现PCR定量检测转基因含量与测定样品转基因含量之间呈高度正相关。研究获得的转基因玉米2A-5转化体特异性PCR检测方法及定量qRT-PCR、ddPCR检测方法具有较高的特异性、准确性和灵敏度，是今后准确、高效检测2A-5及其产品的有效方法之一，同时也为我国转基因生物安全监管提供了良好的技术支撑。

关键词: 转基因玉米, 2A-5, PCR检测, 定量检测

Abstract: According to the flanking sequence information of transgenic maize 2A-5, the optimal specific primer and TaqMan probe combination 2A-5-5-QF8, 2A-5-5-QR8, 2A-5-5-QP8 were designed and screened. The reaction system of the combinations was optimized as well, and the specific PCR detection method of transgenic event 2A-5 was established. The quantitative detection method of transgenic maize 2A-5 was studied by qRT-PCR and ddPCR. It was found that the combination of specific primers and TaqMan probe could complete the quantitative detection well. There was a high positive correlation between the content of transgenic components obtained by PCR reaction and the content of the sample. The specific PCR detection method and quantitative qRT-PCR and ddPCR detection method of transgenic maize 2A-5 in this study can detect 2A-5 with high specificity, accuracy and sensitivity, which is one of the effective methods to detect 2A-5 and its products accurately and efficiently in the future, and it also can provide good technical support for the safety supervision of genetically modified organisms in China.

Key words: genetically modified maize, 2A-5, PCR detection, quantitative detection

武文艳,刘新香,周秒依,刘金香,刘亚. 转基因玉米2A-5特异性PCR方法的建立[J]. 生物技术进展, 2020, 10(4): 363-370.

WU Wenyan^§, LIU Xinxiang^§, ZHOU Miaoyi^*, LIU Jinxiang, LIU Ya. Establishment of Event-specific PCR Detection Method of Transgenic Maize Line 2A-5[J]. Curr. Biotech., 2020, 10(4): 363-370.

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转基因玉米2A-5特异性PCR方法的建立

Establishment of Event-specific PCR Detection Method of Transgenic Maize Line 2A-5

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