Agrobacterium tumefaciens T-DNA Integration and Gene Targeting in Arabidopsis thaliana Non-Homologous End-Joining Mutants
Figure 1
Molecular analysis of the T-DNA insertion lines. (a) Genomic organization of the AtKu70 and AtKu80 loci in the Atku70 and Atku80 mutants. Inserted T-DNA and pROK2 vector part of the binary vector are indicated. Exons are shown as grey boxes, 3′ and 5′ UTRs are shown as white boxes, and introns are shown as lines. The primers used for genotyping and Q-RT-PCR analysis are indicated. The probe and the restriction enzyme digestion sites (H for HindIII) used for Southern blot analysis are also indicated. The sequence of the T-DNA-genomic DNA junctions are shown: genomic DNA (italic) in dark grey boxes, T-DNA insertions in light grey boxes and filler DNA not boxed. (b) Southern blot analysis of the Atku70 and Atku80 mutants. DNA was digested with HindIII and separated in 0.7% agarose gel along a dig-labeled Lambda EcoRI/HindIII marker (lane M), blotted, and hybridized with a DIG-labelling T-DNA probe. The bands with the expected size are indicated with an asterisk.
