Figure 6.

Lack of Rgf1p decreases spontaneous HR activity. (A) An example of a recombination reporter gene ura4::kanMX6 (RDUX) with a 200-bp tandem duplication (RDUX200(+)) and its reversion product (ura4+). (lower panel) Spontaneous recombination rate in wild-type, rgf1Δ, rho1-596 and rad51Δ cells was measured by using the RDUX200 reporter as described in the Materials and Methods. The experiments were performed three times and error bars represent 95% CI. Statistical significance was calculated using two-tailed unpaired Student's t test. The asterisk indicates that the rgf1Δ and rho1-596 value are significantly different from that of the wild type. **P < 0.01 and ***P < 0.001. (B) A value for NHEJ efficiency is represented by an L/C ratio which was generated by dividing the transformation frequency obtained with linear plasmid DNA (L) with that obtained with covalently closed circular DNA (C) (uncut pFY20), which normalized any strain to strain variance in the transformation efficiency (rather than NHEJ efficiency). Values are the means from three independent experiments, and S.D. Bars are indicated. Statistical significance was calculated using two-tailed unpaired Student's t test; ns = non-significant. ***P < 0.001. (Lower panel) Serial dilutions (1, 0.5, 0.25, 0.025, 0.0025 and 0.00025) of the indicated strains were spotted onto rich YES plates with or w/o Phl. Colony formation was analyzed after 3 days at 28°C.