Article

Euphytica

, Volume 123, Issue 1, pp 75-84

First online:

Identification and characterization of T-DNA inserts by T-DNA fingerprinting

  • I. TheunsAffiliated withDepartment of Plant Genetics and Breeding, Center of Agricultural Research-Gent
  • , P. WindelsAffiliated withDepartment of Plant Genetics and Breeding, Center of Agricultural Research-Gent
  • , S. De BuckAffiliated withDepartments of Molecular Genetics and Plant Genetics, Flanders Interuniversity Institute for Biotechnology, University of Gent, K.L
  • , A. DepickerAffiliated withDepartments of Molecular Genetics and Plant Genetics, Flanders Interuniversity Institute for Biotechnology, University of Gent, K.L
  • , E. Van BockstaeleAffiliated withDepartment of Plant Genetics and Breeding, Center of Agricultural Research-GentDepartment of Plant Production, University of Gent
  • , M. De LooseAffiliated withDepartment of Plant Genetics and Breeding, Center of Agricultural Research-Gent Email author 

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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.

AFLP anchored PCR GMO detection T-DNA integration