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. 2005 Dec 27;102(52):19231–19236. doi: 10.1073/pnas.0506437103

Fig. 2.

Fig. 2.

The KU80 protein binds to and recombines dsT-DNA molecules in planta. (a) PCR analysis of T-DNA molecules immunoprecipitated and PCR-amplified as described (28) with antibodies against a His-tagged KU80 protein and using primers specific to T-DNA (Left), binary plasmid (center) and Agrobacterium VirE2 (Right). Lanes 1 (a and b) Precipitated extract treated with anti-His antibodies; lanes 2 (a and b) mock-precipitated extract to which no antibody was added; lanes 3 (a and b) untreated total DNA extract; lane 4 (Center) amplification of plasmid DNA and lane 4 (Right) amplification of bacterial DNA. (b) T-DNA to T-DNA recombination assay. Wild-type (Left) and KU80-overexpressing (Right) root segments were cotransformed with two binary plasmids, each carrying a partial GUS expression cassette (i.e., promoter-GUSint and intGUS-terminator). T-DNA to T-DNA recombination and expression of the GUS reporter gene were monitored by 5-bromo-4-chloro-3-indolyl β-d-glucuronide (X-Gluc) staining. Arrows (Left) indicate the presence of GUS-stained tissue in wild-type root segments.