植物源性转基因食品PCR检测技术研究

PCR for identification of genetically modified plants in food

〔目的〕建立简便、快速、灵敏、特异的植物源性转基因食品DNA检测技术。〔方法〕针对转基因植物技术中载体上常用花椰菜花叶病毒的 3 5S启动子和根农杆菌的NOS终止子特异基因序列 ,设计合成特异的引物对 3 5s -f/ 3 5s-r、Nos -f/Nos -r ,利用基因PCR扩增技术检测植物源性转基因食品。〔结果〕引物对 3 5s -f/ 3 5s -r、Nos -f/Nos -r分别成功从转基因大豆中扩增出 195bp和 180bp特异的DNA带。采用引物对 3 5s -f/ 3 5s -r进行PCR时检测灵敏度为每反应 10 4分子 ;采用Nos -f/Nos -r时检测灵敏度为 2× 10 4分子。最低可以对 0 .1μg转基因大豆进行检测。〔结论〕基于 3 5S启动子和NOS终止子基因序列的PCR扩增技术是一种简便、快速、灵敏。

〔Objective〕 To establish a simple,rapid,sensitive and specific method for identification of genetically modified plants in food.〔Methods〕 Based on the sequences of 35s promoter from CaMV and Nos terminator from Agrubacterium tumefaciens,2 primer pairs:35s-f/35s-r and Nos-f/Nos-r were designed to develop a PCR assay for identification of genetically modified plants in food.〔Results〕A 195 bp specific DNA fragment as predicted was produced from genetically modified bean seeds genome DNA by PCR assay using the primer pair 35s-f/35s-r and a 180 bp specific DNA fragment using the primer pair Nos-f/Nos-r.The sensitivity of PCR assay using primer pair 35s-f/35s-r was 10 4 copies per reaction and using primer pair Nos-f/Nos-r 2×10 4 copies per reaction.Both assays could detect our as low as 0.034 ng genome DNA from 0.1 mg genetically modified bean seeds.〔Conclusions〕 The PCR assays based on the sequences of 35s promoter from CaMV and Nos terminator from Agrubacterium tumefaciens are simple,rapid,sensitive,specific and reliable for identification of genetically modified plants in food.