Recovery of Agrobacterium tumefaciens T-DNA molecules from whole plants early after transfer

Cell. 1989 Jun 2;57(5):847-57. doi: 10.1016/0092-8674(89)90799-x.

Abstract

A system for the analysis of independent T-DNA transfer events from Agrobacterium to plants is described. The complete T-DNA except for the 25 bp border sequences was replaced by one genome of a plant virus so that upon transfer to the plant, a viable replicon is produced by circularization. Rescue of virus from such infected plants allowed analysis of DNA sequences at or close to the ends of T-DNA molecules. A rather conserved right border remnant of three nucleotides was found, whereas the sequences remaining at the left end were more variable. A point deletion in the left 25 bp sequence results in even less precise processing at the left end. In addition, many rescued T-DNA molecules carry small direct repeats between the joined T-DNA ends; linear T-DNA molecules are therefore transported to the plant.

MeSH terms

  • Base Sequence
  • DNA, Bacterial / genetics
  • DNA, Bacterial / isolation & purification*
  • Genes, Bacterial*
  • Molecular Sequence Data
  • Plants / genetics
  • Plants / microbiology*
  • Plasmids
  • Rhizobium / genetics*
  • Transfection

Substances

  • DNA, Bacterial