TY - JOUR T1 - An innovative and integrated approach based on DNA walking to identify unauthorised GMOs JF - Food Chemistry Y1 - 2014 A1 - Marie-Alice Fraiture A1 - Philippe Herman A1 - Taverniers, Isabel A1 - De Loose, Marc A1 - Deforce, Dieter A1 - Nancy Roosens KW - DNA walking KW - GM rice KW - GMO detection KW - Integrated approach KW - pCAMBIA vector KW - Quantitative real-time PCR KW - Transgene flanking region KW - Unauthorised GMO AB -

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®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.

VL - 147 M3 - https://doi.org/10.1016/j.foodchem.2013.09.112 ER -