A quick method to estimate the T-DNA copy number in transgenic plants at an early stage after transformation, using inverse PCR

Plant Mol Biol. 1991 Jul;17(1):151-3. doi: 10.1007/BF00036819.

Abstract

Agrobacterium-mediated transformation of plants is known to result in transgenic plants with a variable number of integrated T-DNA copies. Our aim was to obtain transgenic tobacco plants containing one integrated T-DNA copy per genome. Therefore, a quick method was developed to estimate the T-DNA copy number of young transgenic plantlets within 10 weeks after transformation. Inverse polymerase chain reaction (IPCR) was used to amplify junction fragments, i.e. plant genomic DNA sequences flanking the known T-DNA sequences.

MeSH terms

  • Base Sequence
  • DNA, Bacterial / analysis*
  • Molecular Sequence Data
  • Plants, Toxic*
  • Polymerase Chain Reaction / methods*
  • Rhizobium / genetics
  • Tobacco / genetics*
  • Transformation, Genetic*

Substances

  • DNA, Bacterial
  • T-DNA