Insights into recognition of the T-DNA border repeats as termination sites for T-strand synthesis by Agrobacterium tumefaciens.
The recognition of the T-DNA left border (LB) repeat is affected by its surrounding sequences. Here, the LB regions were further characterized by molecular analysis of transgenic plants, obtained after Agrobacterium tumefaciens-mediated transformation with T-DNA vectors that had been modified in this LB region. At least the 24-bp LB repeat by itself was insufficient to terminate the T-strand synthesis. Addition of the natural inner and/or outer border regions to at least the LB repeat, even when present at a distance, enhanced the correct recognition of the LB repeat, reducing the number of plants containing vector backbone sequences. In tandem occurrence of both the octopine and nopaline LB regions with their repeats terminated the T-strand synthesis most efficiently at the LB, yielding a reproducibly high number of plants containing only the T-DNA. Furthermore, T-strand synthesis did not terminate efficiently at the right border (RB) repeat, which might indicate that signals in the outer RB region inhibit the termination of T-strand synthesis at the RB repeat.
Podevin
Nancy
N
Department of Plant Systems Biology, Flanders Interuniversity Institute for Biotechnology (VIB), Ghent University, Technologiepark 927, B-9052 Gent, Belgium.
De Buck
Sylvie
S
De Wilde
Chris
C
Depicker
Anna
A
D016960:Agrobacterium tumefaciens / Q000235:genetics* / Q000378:metabolism*; D017360:Arabidopsis / Q000235:genetics* / Q000378:metabolism; D004269:DNA, Bacterial / Q000235:genetics* / Q000378:metabolism; D018744:DNA, Plant / Q000096:biosynthesis*; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D012091:Repetitive Sequences, Nucleic Acid
D004269:DNA, Bacterial; D018744:DNA, Plant; C036483:T-DNA