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Development of a standard, validated procedure for the isolation of transgene flanking regions in GM crops and detailed analysis of transgene insertion

Project Code: G03026;
Publication:

Cullen, D., Harwood, W. A., Smedley, M. A., Davies, H., & Taylor, M. (2011) Comparison of DNA walking methods for isolation of transgene-flanking regions in GM potato. Molecular Biotechnology, 49(1), 19-31, published online doi: 10.1007/s12033-010-9371-5

19/04/2011

John Innes Centre
Harwood, W ;
Scottish Crop Research Institute
Taylor, M

The isolation and analysis of transgene flanking regions forms a key component of the molecular analysis and safety assessment of GM plants.

It is important to identify exactly where the additional DNA (transgene) has been inserted in a GM plant as this may highlight whether a plant gene has been disrupted or whether there are rearrangements of the DNA at the site of insertion that may be of concern; for example if a new open reading frame has been created that may be expressed in the plant. Although there are a number of methods available that allow the transgene junction region to be isolated, in general these methods are complex and have been used only with the insertion of specific transgenes or with specific plant species.

The aim of this project was to develop standard and validated PCR-based methods to allow the routine isolation and characterisation of the transgene junction regions in a range of different GM crops with diverse transgene insertions of different complexity.

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