Enzymatic synthesis of deoxyribo-oligonucleotides of defined sequence. Properties of the enzyme

Abstract

A modified purification is described for an enzyme, from Escherichia coli B, which polymerizes deoxyribonucleoside-5^′ diphosphates. Under appropriate conditions, the enzyme will add a single deoxyribonucleotide residue to a deoxyribo-oligo-nucleotide primer. At all stages, the enzyme activity copurified with the activity which will polymerize adenosine-5^′ diphosphate (polynucleotide phosphorylase). Studies of heat stability, the effect of various temperatures of reaction and of disc gel electrophoresis failed to provide evidence that the two activities are separable.