Random Mutagenesis Using 2-Amino-9-(2-Deoxy-β-D-Ribofuranosyl)Purine-5′-Triphosphate and the Polymerase Chain Reaction

Abstract

Base analogues offer an attractive method for mutagenising DNA in combination with the polymerase chain reaction (PCR). We have synthesised the 5′-triphosphate-2′-deoxyribosyl derivative of 2-aminopurine (dAPTP), one of the first base analogues to be used for mutagenesis, and examined its utility in PCRs. An E. coli amber suppressor gene, supF, was used as a template for mutagenesis. The analogue induced exclusively transition mutations, but at a low frequency, consistent with its weak mutagenicity in vivo.