转基因产品中抗除草剂基因的SYBR®绿色qPCR筛选方法

Zhen Zhen , Lv Wei , Tang Zhi-fen , Liu Ying , Ao Jin-xia , Yuan Xiao-han , Zhang Ming-hui , Qiu You-wen , Gao Xue-jun
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引用次数: 3

摘要

转基因生物(GMOs)作为食品的使用越来越广泛。欧盟已经实施了一套非常严格的程序来批准种植、进口和/或使用转基因生物作为食品或食品成分。因此,检测转基因生物的分析方法是必要的,以便核实是否符合标签要求。目前对转基因加工产品的有效筛选方法很少。CP4-EPSPS、BAR和PAT三种抗除草剂基因是商品化转基因大豆、玉米和水稻中常用的外源基因。本研究建立了一种新的SYBR®Green qPCR筛选方法,可同时检测3个外源抗除草剂基因和1个内源抗除草剂基因。采用该方法对7种具有代表性的加工产品(大豆卵磷脂、大豆蛋白粉、巧克力饮料、婴儿米粉、玉米蛋白粉、玉米淀粉和玉米果酱)进行了检测,获得了所有加工产品的内源基因扩增子和转基因片段,灵敏度为0.1%。结果表明,SYBR®Green qPCR筛选方法适用于转基因大豆、玉米和水稻加工产品的定性检测。
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SYBR® Green qPCR Screening Methods for Detection of Anti-herbicide Genes in Genetically Modified Processed Products

The use of genetically modified organisms (GMOs) as food products becomes more and more widespread. The European Union has implemented a set of very strict procedures for the approval to grow, import and/or utilize GMOs as food or food ingredients. Thus, analytical methods for detection of GMOs are necessary in order to verify compliance with labelling requirements. There are few effective screening methods for processed GM (genetically modified) products. Three anti-herbicide genes (CP4-EPSPS, BAR and PAT) are common exogenous genes used in commercialized transgenic soybean, maize and rice. In the present study, a new SYBR® Green qPCR screening method was developed to simultaneously detect the three exogenous anti-herbicide genes and one endogenous gene in a run. We tested seven samples of representative processed products (soya lecithin, soya protein powder, chocolate beverage, infant rice cereal, maize protein powder, maize starch, and maize jam) using the developed method, and amplicons of endogenous gene and transgenic fragments were obtained from all the processed products, and the sensitivity was 0.1%. These results indicated that SYBR® Green qPCR screening method was appropriate for qualitative detection of transgenic soybean, maize and rice in processed products.

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