多重荧光PCR同时检测转基因成分35S和Nos方法的建立  

Multiplex Fluorescence PCR Method for Detecting Transgenic Component 35S and Nos Simultaneously

作　　者：刘光明[1,2] 李庆阁[1] 王群力[3] 梁基选[1] 陈伟铃[3] 栾国彦[1] 苏文金[1] 

机构地区：[1]厦门大学生命科学学院 [2]厦门出入境检验检疫局,福建厦门361012 [3]厦门出入境检验检疫局

出　　处：《厦门大学学报（自然科学版）》2002年第4期493-497,共5页Journal of Xiamen University：Natural Science

基　　金：厦门市科技计划资助项目 (35 0 2Z2 0 0 110 9)

摘　　要：根据商品化转基因作物中常用的花椰菜花叶病毒启动子 (CaMV 35S)和根癌农杆菌终止子 (Nos)的序列特点 ,设计并合成了两对引物和相对应的荧光双链探针 ,建立一种应用荧光双链探针的多重荧光PCR同时检测转基因成分 35S启动子和Nos终止子的方法 .并利用该方法对马铃薯、大豆、玉米、甜椒、番茄等 11份实物样品进行了检测 ,其中有 5份样品结果阳性 .结果表明所建立的多重荧光PCR方法能同时检测出 35S和Nos双组分 ,较常规PCR技术更为简便、快速、准确 ,有很好的应用前景 .The article is to establish Multiplex Fluorescence PCR (MF PCR) method for detecting transgenic component 35S promoter derived from Cauliflower Mosaic Virus and Nos terminator derived from Agrobacterium tumefaciens simultaneously. 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 were designed and synthesized for detection of 35S & Nos simultaneously with MF PCR in a tube. 11 samples were tested with this method. The results showed that 5 samples were positive, 6 samples were negative. The described methods enabled a sensitive?specific?simple and accurate detection of genetically modified food and thus provide a useful tool for routine analysis of raw and processed food products.

关 键 词：多重荧光PCR 35S NOS 转基因作物 荧光双链探针 花椰菜花叶病毒启动子 极癌农杆菌终止子 检测方法 

分 类 号：Q943.2[生物学—植物学]