Synthesis of Poly[d(A‐s4T) · d(A‐s4T)] by Bacillus subtilis DNA Polymerase
Definition. A260 unit, the quantity of material contained in 1 ml of a solution which has an absorbance of 1 at 260 nm, when measured in a 1‐cm pathlength cell.
Abstract
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.
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