Table I.

Efficiency of homologous recombination in Rad54 ATPase-defective cells

Genotype Targeting efficiency at Rb locus 
Rad54WT-GFP/- 31.9% (16 out of 46) 
Rad54K189R-GFP/- 0.95% (1 out of 105) 
Rad54K189A-GFP/- 1.2% (1 out of 81) 
Rad54−/− <1% (0 out of 82) 
Genotype Targeting efficiency at Rb locus 
Rad54WT-GFP/- 31.9% (16 out of 46) 
Rad54K189R-GFP/- 0.95% (1 out of 105) 
Rad54K189A-GFP/- 1.2% (1 out of 81) 
Rad54−/− <1% (0 out of 82) 

ES cells with the indicated genotypes were electroporated with a linearized Rb-Hyg construct (Niedernhofer et al., 2001). Hygromycin-resistant clones were expanded, genomic DNA was purified, and samples were subjected to DNA blot analysis to distinguish between randomly and homologously integrated events. Values indicate the percentage of clones that contain the homologously integrated targeting construct relative to the total number of clones analyzed. Absolute numbers are indicated in parentheses. The differences in recombination efficiency between Rad54WT-GFP/− cells and cells of all other genotypes listed are significant (P > 0.001), whereas the difference between the mutant genotypes is not.

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