食品科学 ›› 2002, Vol. 23 ›› Issue (5): 94-99.

• 分析检测 • 上一篇    下一篇

PCR方法检测食品中的转基因成分35S和NOS的研究

 刘光明, 苏文金, 栾国彦, 宋思扬, 梁基选   

  1. 厦门出入境检验检疫局,厦门大学生命科学学院
  • 出版日期:2002-05-15 发布日期:2012-02-13

PCR Method for Assaying Transgenic Component 35S and NOS in Foods

 LIU  Guang-Ming, SU  Wen-Jin, LUAN  Guo-Yan, SONG  Si-Yang, LIANG  Ji-Xuan   

  • Online:2002-05-15 Published:2012-02-13

摘要: 根据转基因农作物中最常用的花椰菜叶病毒启动子(CaMV35S)和根癌农杆菌终止子(NOS)的序列,设计并合成了两对不同的引物和相对应的两种荧光双链探针(FDCP),分别建立了常规PCR、应用FDCP的新型实时荧光PCR检测转基因成分35S启动子和NOS终止子的方法。利用该套方法对冷冻马铃薯条、大豆、冷冻玉米棒、甜椒、番茄等实物样品进行了检测,发现11份样品中有5份检出35S启动子、NOS终止子,6份样品结果为阴性。结果表明我们建立的两种PCR方法均能有效检测出35S和NOS片段,其中常规PCR方法具有灵敏度高、特异性好的特点;应用FDCP的新型实时荧光PCR方法则更为简便、快速、准确,有很好的应用前景和研究价值。

关键词: 转基因, PCR, 荧光PCR, 荧光双链探针

Abstract: The article was is to establish PCR method for assaying transgenic component 35S promoter derived from Cauliflower Mosaic Virus and NOS terminator derived from Agrobacterium tumefaciens in foods.According to the specific sequence of 35S and NOS Which have been used in transgenic crops frequently,two pairs of primers and two pairs of corresponding Fluorophore double-chain probes (FDCP) were designed to amplify parts of 35S promoter and NOS terminator,to allow for general screening of foods.PCR & Fluorescence PCR(FPCR)methods were established for the assay of transgenic component 35S and NOS.11 samples were tested with PCR & FPCR.The results showed that 5 samples were positive,6 samples were negative.The described methods enabled a sensitive、specific、simple and accurate assay of transgenic component and thus provided & a useful tool for routine analysis of raw and processed food products.

Key words: Transgenic , PCR , Fluorescence PCR , Fluorophore double-chain probe