An innovative and integrated approach based on DNA walking to identify unauthorised GMOs

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• 作者：Marie-Alice Fraiture ; Philippe Herman ; Isabel Taverniers ; Marc De Loose ; Dieter Deforce ; Nancy H. Roosens
• 关键词：Unauthorised GMO ; GM Rice ; pCAMBIA vector ; DNA walking ; Transgene flanking region ; GMO detection ; Quantitative real-time PCR ; Integrated approach
• 刊名：Food Chemistry
• 出版年：15 March, 2014
• 年：2014
• 卷：147
• 期：Complete
• 页码：60-69
• 全文大小：1812 K

文摘

In the coming years, the frequency of unauthorised genetically modified organisms (GMOs) being present in the European food and feed chain will increase significantly. Therefore, we have developed a strategy to identify unauthorised GMOs containing a pCAMBIA family vector, frequently present in transgenic plants. This integrated approach is performed in two successive steps on Bt rice grains. First, the potential presence of unauthorised GMOs is assessed by the qPCR SYBR庐Green technology targeting the terminator 35S pCAMBIA element. Second, its presence is confirmed via the characterisation of the junction between the transgenic cassette and the rice genome. To this end, a DNA walking strategy is applied using a first reverse primer followed by two semi-nested PCR rounds using primers that are each time nested to the previous reverse primer. This approach allows to rapidly identify the transgene flanking region and can easily be implemented by the enforcement laboratories.