Enzymatic synthesis of deoxyribonucleic acid. I. Preparation of substrates and partial purification of an enzyme from Escherichia coli.

In the foregoing paper (l), a procedure was described for purifying an enzyme (“polymerase”) from Escherichia coli which catalyzes the incorporation o f deoxyribonucleotides into DNA. The purpose o f this report is to describe the requirements for the reaction, the net synthesis o f DNA, and some chemical features of the synthesized DNA. The liberation of inorganic pyrophosphate as a consequence o f the reaction and studies on the participation of inorganic pyrophosphate in reversing the polymerization will also be presented.