Identification and characterization of T-DNA inserts by T-DNA fingerprinting
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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.
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- Identification and characterization of T-DNA inserts by T-DNA fingerprinting
Volume 123, Issue 1 , pp 75-84
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- anchored PCR
- GMO detection
- T-DNA integration
- Industry Sectors
- Author Affiliations
- 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