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Synthesis of poly (d(A-s 4 T)-d(A-s 4 T)) by Bacillus subtilis DNA polymerase.

Two bacterial DNA polymerases lacking the 5′-exonuclease, isolated from Escherichia coli and Bacillus subtilis are compared with respect to their capability to synthesize the nucleoside analong containing polydeoxynucleotide poly[d(A-s4T) · d(A-s4T)] extensively. Only the enzyme from B. subtilis can use this biopolymer as template for further synthesis, whereas the DNA polymerase A of E. coli requires poly[d(A-T) · d(A-T)] to copy. A method for preparation of pure poly[d(A-s4T) · d(A-s4T)] is described that can be scaled up at will. Proof is given by nucleoside analysis after enzymatic digestion that the polymer consists of equal amounts of deoxyadenosine and 4-thiothymidine nucleotides. Absorption coefficients of poly[d(A-sT) · d(A-s4T)] were determined by two independent methods.