食品科学

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实时荧光聚合酶链式反应检测转基因小麦B73-6-1、B72-8-11b和B102-1-2品系

曹际娟1,徐君怡1,曹冬梅1,张丕桥1,2,栾凤侠3,刘 洋1   

  1. 1.辽宁出入境检验检疫局,辽宁 大连 116001;2.辽宁师范大学生命科学学院,辽宁 大连 116029;
    3.黑龙江出入境检验检疫局,黑龙江 哈尔滨 150001
  • 出版日期:2014-04-25 发布日期:2014-05-13
  • 基金资助:

    国家转基因重大专项(2013ZX08012-001)

A Real-Time Polymerase Chain Reaction Method to Detect Genetically Modified Wheat Strains B73-6-1, B72-8-11b and B102-1-2

CAO Ji-juan1, XU Jun-yi 1, CAO Dong-mei1, ZHANG Pi-qiao1,2, LUAN Feng-xia3, LIU Yang1   

  1. 1. Liaoning Entry-Exit Inspection and Quarantine Bureau, Dalian 116001, China;
    2. College of Life Science, Liaoning Normal University, Dalian 116029, China;
    3. Heilongjiang Entry-Exit Inspection and Quarantine Bureau, Harbin 150001, China
  • Online:2014-04-25 Published:2014-05-13

摘要:

目的:建立实时荧光聚合酶链式反应检测转基因小麦B73-6-1、B72-8-11b和B102-1-2品系的鉴定方法。方 法:以转基因小麦B73-6-1、B72-8-11b和B102-1-2为研究对象,在转基因小麦外源片段与小麦染色体重组的边界序 列分别设计引物和探针,以其他多种转基因产品和非转基因小麦为对照进行特异性实验,以B73-6-1样品模拟制备 10 个含量梯度的添加样品进行灵敏度实验。结果:本研究设计的引物和探针具有很好的品系鉴定特异性,检测灵敏 度可达到0.01%(m/m)。结论:该方法特异性好、灵敏度高,可快速、准确地鉴定转基因小麦B73-6-1、B72-8-11b 和B102-1-2品系。

关键词: 转基因小麦, B73-6-1, B72-8-11b, B102-1-2, 品系鉴定, 实时荧光聚合酶链式反应

Abstract:

Aim: To establish a real-time PCR method for the identification of transgenic wheat strains B73-6-1, B72-8-11b
and B102-1-2. Methods: Specific primers and probes were designed according to the flanking sequences of exogenous
fragments of transgenic wheat and wheat chromosomes. The specificity of the developed method was validated by using
it to detect a variety of other genetically modified crops and non-genetically modified wheat strains. B73-6-1 was used
to prepare 10 content gradients to test the sensitivity. Results: The primers and probes designed showed good detection
specificity for transgenic wheat strains, with its sensitivity reaching 0.01% (m/m). Conclusion: A real-time PCR method for
the identification of three genetically modified wheat strains has been established. This method is of high specificity, high
reproducibility, rapid identification and excellent accuracy for the identification of genetically modified wheat strains B73-6-1,
B72-8-11b and B102-1-2.

Key words: genetically modified wheat, B73-6-1, B72-8-11b, B102-1-2, event specific detection, real-time polymerase chain reaction (PCR)

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