Recovery of Agrobacterium tumefaciens T-DNA molecules from whole plants early after transfer.
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.
Bakkeren
G
G
Friedrich Miescher-Institut, Basel, Switzerland.
Koukolíková-Nicola
Z
Z
Grimsley
N
N
Hohn
B
B
D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D010944:Plants / Q000235:genetics / Q000382:microbiology*; D010957:Plasmids; D012231:Rhizobium / Q000235:genetics*; D014162:Transfection
D004269:DNA, Bacterial