Deviating T-DNA transfer from Agrobacterium tumefaciens to plants.
We analyzed 29 T-DNA inserts in transgenic Arabidopsis thaliana plants for the junction of the right border sequences and the flanking plant DNA. DNA sequencing showed that in most lines the right border sequences transferred had been preserved during integration, corroborating literature data. Surprisingly, in four independent transgenic lines a complete right border repeat was present followed by binary vector sequences. Cloning of two of these T-DNA inserts by plasmid rescue showed that in these lines the transferred DNA consisted of the complete binary vector sequences in addition to the T-region. On the basis of the structure of the transferred DNA we propose that in these lines T-DNA transfer started at the left-border repeat, continued through the vector part, passed the right border repeat, and ended only after reaching again this left-border repeat.
van der Graaff
E
E
Institute of Molecular Plant Sciences, Leiden University, Clusius Laboratory, Netherlands.
den Dulk-Ras
A
A
Hooykaas
P J
PJ
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D017931:DNA Primers; D004269:DNA, Bacterial / Q000737:chemistry / Q000378:metabolism*; D018744:DNA, Plant / Q000737:chemistry / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000378:metabolism; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified; D010957:Plasmids; D016133:Polymerase Chain Reaction; D014162:Transfection / Q000379:methods
D017931:DNA Primers; D004269:DNA, Bacterial; D018744:DNA, Plant; D004277:DNA, Single-Stranded; C036483:T-DNA