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. 2001 Apr 10;98(9):4978–4983. doi: 10.1073/pnas.081088798

Figure 3.

Figure 3

DNA replication can be restored by adding purified Polɛ back to the depleted extracts. (A) Purification of Xenopus Polɛ from egg extracts. An elution profile of DNA polymerase activity from Mono Q column (Top), immunoblots with Polɛ p60 or Polα p70 antibodies (Middle), and a silver-stained protein gel (Bottom) of the corresponding fractions are shown. Arrowheads indicate the positions of the catalytic subunit (p260) and p60 of Polɛ, respectively. (B) Restoration of DNA replication by the addition of purified Polɛ back to Polɛ-depleted extracts. The products from the reactions (70-min incubation) with mock- or Polɛ-depleted extracts (14 μl each) supplemented with 1 μl each of control buffer or each Mono Q fraction containing Polɛ (fraction 28 in A), Polα (fraction 23 in A), or free p60 (fraction 21 in A) were analyzed as in Fig. 2C. The position of the origin for gel electrophoresis (ori) is indicated on the right. The sizes of marker DNA are on the left.