Abstract
[Introduction] Formation of a phosphorus-oxygen bond between phosphate and hydroxyl termini of DNA in aqueous solution requires chemical activation of the phosphate for nucleophilic substitution and positioning of the hydroxyl for attack on the activated phosphate in competition with water. This esterification reaction is accomplished enzymatically by DNA ligases, which utilize energy from an ATP or NAD cofactor to activate the phosphates. We report a nonenzymatic approach to ligation of double-helical DNA employing a single-stranded template to align two duplex strand termini in a local triple helix (Figure I). A triple-stranded complex is formed by association of a pyrimidine oligodeoxyribonucleotide in the major groove of the Watson-Crick duplex with sequence specificity derived from Hoogsteen hydrogen bonding. Juxtaposition of the two DNA termini by a guide sequence in a triple helix, accompanied by chemical activation of the terminal phosphates, promotes ligation of the double-helical DNA.
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