Kitagawa Mamiko, Nakamura Kosuke, Kondo Kazunari, Ubukata Shoji, Akiyama Hiroshi
Research and Development Division, KAGOME Co., Ltd.
Shokuhin Eiseigaku Zasshi. 2014;55(6):247-53. doi: 10.3358/shokueishi.55.247.
The contamination of processed vegetable foods with genetically modified tomatoes was investigated by the use of qualitative PCR methods to detect the cauliflower mosaic virus 35S promoter (P35S) and the kanamycin resistance gene (NPTII). DNA fragments of P35S and NPTII were detected in vegetable juice samples, possibly due to contamination with the genomes of cauliflower mosaic virus infecting juice ingredients of Brassica species and soil bacteria, respectively. Therefore, to detect the transformation construct sequences of GM tomatoes, primer pairs were designed for qualitative PCR to specifically detect the border region between P35S and NPTII, and the border region between nopaline synthase gene promoter and NPTII. No amplification of the targeted sequences was observed using genomic DNA purified from the juice ingredients. The developed qualitative PCR method is considered to be a reliable tool to check contamination of products with GM tomatoes.
通过使用定性PCR方法检测花椰菜花叶病毒35S启动子(P35S)和卡那霉素抗性基因(NPTII),对加工蔬菜食品中转基因番茄的污染情况进行了调查。在蔬菜汁样品中检测到了P35S和NPTII的DNA片段,这可能分别是由于感染十字花科植物汁成分的花椰菜花叶病毒基因组和土壤细菌的污染所致。因此,为了检测转基因番茄的转化构建体序列,设计了引物对用于定性PCR,以特异性检测P35S和NPTII之间的边界区域,以及胭脂碱合酶基因启动子和NPTII之间的边界区域。使用从汁成分中纯化的基因组DNA未观察到目标序列的扩增。所开发的定性PCR方法被认为是检查产品是否被转基因番茄污染的可靠工具。