Fig. 5.

Activity of dormant origin ARS301 in hst3 mutant. Genomic DNA was prepared from hst3Δ (YIC281) and wild-type (YJT503) strains. After NdeI digestion, replication intermediates were separated by 2D electrophoresis, blotted and probed with a fragment containing VBA3 and ARS301, indicated by the black bar below the diagram of the NdeI fragment containing ARS301. The fragment from the balancer chromosome III is 4.8 kb; the extra NdeI site present on the 5ORIΔ-ΔR fragment shortens the ARS301 fragment to 4.1 kb. The ARS301 probe also hybridized to a 7.1-kb NdeI fragment on chromosome XI containing the VBA5 gene. The arrows point to the bubble-shaped intermediates arising from the 5ORIΔ-ΔR fragment