Deoxyribonucleic Acid Synthesis in Bacteriophage SP01-infected Bacillus subtilis

Abstract

Abstract A new DNA polymerase has been purified 1000-fold from Bacillus subtilis infected with bacteriophage SP01. It consists of a single polypeptide chain of approximately 122,000 molecular weight. The enzyme requires deoxyribonucleoside 5'-triphosphates, Mg2+, β-mercaptoethanol, and single stranded DNA templates for activity. Bihelical DNA which has been partially denatured serves as the preferred template in in vitro reactions. Although 5-hydroxymethyluracil replaces thymine in SP01 DNA, deoxynucleotide analogues of 5-hydroxymethyldeoxyuridine 5'-triphosphate can serve as efficient substrates for the phage polymerase. The purified enzyme is free of endonuclease activity.