Detection and identification of multiple genetically modified events using DNA insert fingerprinting
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Current screening and event-specific polymerase chain reaction (PCR) assays for the detection and identification of genetically modified organisms (GMOs) in samples of unknown composition or for the detection of non-regulated GMOs have limitations, and alternative approaches are required. A transgenic DNA fingerprinting methodology using restriction enzyme digestion, adaptor ligation, and nested PCR was developed where individual GMOs are distinguished by the characteristic fingerprint pattern of the fragments generated. The inter-laboratory reproducibility of the amplified fragment sizes using different capillary electrophoresis platforms was compared, and reproducible patterns were obtained with an average difference in fragment size of 2.4 bp. DNA insert fingerprints for 12 different maize events, including two maize hybrids and one soy event, were generated that reflected the composition of the transgenic DNA constructs. Once produced, the fingerprint profiles were added to a database which can be readily exchanged and shared between laboratories. This approach should facilitate the process of GMO identification and characterization.
KeywordsGenetically modified organism detection GMO Anchored PCR Genome walking Fingerprint
Work conducted at the National Measurement Institute, Australia was funded by the Office of the Gene Technology Regulator, Australia. CFIA research was funded by the Canadian Regulatory System for Biotechnology (CRSB). We thank Kate Griffiths, Carole Simard, and André Perron for their constructive review of the manuscript.
- 3.Holst-Jensen A, Rønning SB, Løvseth A, Berdal KG (2003) Anal Bioanal Chem 375:985–993Google Scholar
- 8.Anklam E, Neumann DA (2002) J AOAC Int 85:754–756Google Scholar
- 10.Lipp M, Brodmann P, Pietsch K, Pauwels J, Anklam E (1999) J AOAC Int 82:923–928Google Scholar
- 12.Kok EJ, Aarts HJ, Van Hoef AM, Kuiper HA (2002) J AOAC Int 85:797–800Google Scholar
- 18.Leimanis S, Hamels S, Nazé F, Mbella GM, Sneyers M, Hochegger R, Broll H, Roth L, Dallmann K, Micsinai A, La Paz JL, Pla M, nen-Nieweler C, Papazova N, Taverniers I, Hess N, Kirschneit B, Bertheau Y, Audeon C, Laval V, Busch U, Pecoraro S, Neumann K, Sel S, Van Dijk J, Kok E, Bellocchi G, Foti N, Mazzara M, Moens W, Remacle J, Van Den Eede G (2008) Eur Food Res Technol 227:1621–1632CrossRefGoogle Scholar
- 22.Spertini D, Beliveau C, Bellemare G (1999) Biotechniques 27:308–314Google Scholar
- 26.Akritidis P, Pasentsis K, Tsftaris AS, Mylona PV, Polidoros AN (2008) Electronic J Biotechnol 11:1–199Google Scholar
- 28.GM Crop Database, Agbios. Merrickville, Canada. http://www.agbios.com. Accessed 1 June 2009
- 31.GMO-Watch Report: Genetically Modified Crops: molecular and regulatory facts. BATS. Centre for Biosafety and Sustainability, Zurich, Switzerland. http://www.bats.ch/gmo-watch/. Accessed 1 June 2009