Nonenzymic sequence-specific ligation of double-helical DNA
- Creators
- Luebke, Kevin J.
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Dervan, Peter B.
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.
Additional Information
© 1991 American Chemical Society. Received May 31 , 1991. This work was supported in part by the National Institutes of Health (GM-35724) and the Caltech Consortium in Chemistry and Chemical Engineering (founding members: E. I. du Pont de Nemours and Company, Inc., Eastman Kodak Company, Minnesota Mining and Manufacturing Company, and Shell Oil Company Foundation). We are grateful for a National Research Service Award to K.J .L. from the National Institute of General Medical Science.Additional details
- Eprint ID
- 66948
- Resolver ID
- CaltechAUTHORS:20160510-145140529
- GM-35724
- NIH
- Caltech Consortium in Chemistry and Chemical Engineering
- NIH Predoctoral Fellowship
- Created
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2016-05-19Created from EPrint's datestamp field
- Updated
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2021-11-11Created from EPrint's last_modified field