DNA polymerase A (an alpha-type polymerase) from the ciliate, Tetrahymena pyriformis, has been purified 260,000-fold (40,000 units/mg protein). The polymerase A did not show any heterogeneity in terms of size and charge during purification. Enzymatic properties of the DNA polymerase A remained unchanged during the purification. Two-dimensional gel electrophoresis revealed that in the first dimension (isoelectric focusing agarose gel), the activity of the purified enzyme was focused at around pH 5.5 and that in the second dimension (sodium dodecyl sulfate-polyacrylamide gel), 135,000- and 66,000-dalton polypeptides emerged from the activity peak at a stoichiometric ratio of about 1:3. The native molecular weight of the DNA polymerase A estimated from the stoichiometric subunit ratio approximately coincided with that estimated from gel filtration on Sephadex G-200 under low ionic strength conditions. The present results strongly suggest the existence of a common high-molecular-weight catalytic core subunit in alpha-type polymerases of eukaryotes.