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G03026: Development of a standard, validated procedure for the isolation of transgene flanking regions in GM crops and detailed analysis of transgene insertion

Monday 12 February 2007

This project will involve the refinement and validation of a rapid polymerase chain reaction (PCR) based method for characterisation of junction sequences in a range of genetically modified (GM) crops.

Study Duration : October 2006 to October 2008

Contractor : John Innes Centre, Norwich

Background

The isolation and analysis of transgene flanking regions forms a key component of the molecular analysis and safety assessment of GM plants. Until recently the characterisation of the insertion point for crop plants has been difficult and time consuming. However, work carried out under the G02 programme and other recently published work has resulted in discovery of a rapid PCR based method, which can be applied to the identification of junction sequences.

The characterisation of the insertion point of the introduced transgenic trait will provide information as to whether any functional genes are disrupted and whether any new open reading frames (ORFs) are created resulting in the synthesis of novel proteins. If so, further work can be carried out to determine whether these ORFs may code for potential allergens or toxins.

The project will allow refinement and validation of this method, advancing the state-of-the art technology in this field. The standard validated methods developed could form part of the risk assessment procedure for a large range of GM crops and different types of transgene insertion.

Research Approach

A new methodology will be compared to the existing technology to see if it offers any advantages in throughput, reproducibility and cost. The project will provide standard validated methods for the analysis of junction sequences in a range of GM crops, containing transgene insertions of different complexity. The methods developed will be validated by the analysis of a standard set of GM samples at two different sites. The project will also provide valuable information on the patterns of transgene insertion in a range of GM crops.

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