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Identification and characterization of T-DNA inserts by T-DNA fingerprinting
Authors:I. Theuns  P. Windels  S. De Buck  A. Depicker  E. Van Bockstaele  M. De Loose
Affiliation:(1) Department of Plant Genetics and Breeding, Center of Agricultural Research-Gent, Caritasstraat 21, B-9090 Melle, Belgium;(2) Departments of Molecular Genetics and Plant Genetics, Flanders Interuniversity Institute for Biotechnology, University of Gent, K.L, Ledeganckstraat 35, B-9000 Gent, Belgium;(3) Department of Plant Production, University of Gent, Coupure Links 653, B-9000 Gent, Belgium
Abstract:A T-DNA fingerprinting method is presented based on amplified fragmentlength polymorphism with an anchored polymerase chain reaction step. Thismethod allows discrimination between different T-DNA inserts in stablytransformed plants. The technique was evaluated by analyzing 51 transgenicArabidopsis lines that had been characterized in detail by genomicblotting. Comparison of the obtained fingerprints with the availableintegration information demonstrated that fingerprints were correlated tothe predicted patterns, except for the inverted repeat junctions and forthose inserts with large deletions at the left or right border. Ourexperiments show that by using T-DNA fingerprinting multi-copy transgeniclines can be eliminated efficiently so that the technique can be used toenrich a population of transgenic plants for putative single-copytransformants.
Keywords:AFLP  anchored PCR  GMO detection  T-DNA integration
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