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. 1999 Aug 3;96(16):9130–9135. doi: 10.1073/pnas.96.16.9130

Figure 1.

Figure 1

Temporally regulated origin associations of Mcm7p, Cdc45p, and DNA polymerases α and ɛ. (AD) Cells of strains OAy534 [Mcm7p-HA (A)], OAy617 [Cdc45p-HA (B)], OAy644 [Polα-HA (C)], and OAy618 [Polɛ-HA (D)] were synchronized in G1 phase with α factor and were released at 16°C (t = 0 min). At the indicated time points, chromatin-containing extracts were prepared from formaldehyde cross-linked cells and were immunoprecipitated with anti-HA monoclonal antibody. PCR with primers specific to the ARS305, ARS1, ARS501, and ARS603 chromosomal replication origins and the non-origin sequences 305+8kb and R11 amplified the precipitated DNA for analysis by PAGE and EtBr staining. (E) FACS analysis indicates the DNA content of cells throughout the time course in B. FACS profiles from the experiments in A, C, and D were very similar. (F) Budding index (% Unbudded) of cells throughout each time course in AD was determined and plotted.