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. 1988 May 11;16(9):3721–3738. doi: 10.1093/nar/16.9.3721

Site-directed modification of DNA duplexes by chemical ligation.

N G Dolinnaya 1, N I Sokolova 1, O I Gryaznova 1, Z A Shabarova 1
PMCID: PMC336552  PMID: 3375071

Abstract

The efficiency of chemical ligation method have been demonstrated by assembling a number of DNA duplexes with modified sugar phosphate backbone. Condensation on a tetradecanucleotide template of hexa(penta)- and undecanucleotides differing only in the terminal nucleoside residue have been performed using water-soluble carbodiimide as a condensing agent. As was shown by comparing the efficiency of chemical ligation of single-strand breaks in those duplexes, the reaction rate rises 70 or 45 times if the 3'-OH group is substituted with an amino or phosphate group (the yield of products with a phosphoramidate or pyrophosphate bond is 96-100% in 6 d). Changes in the conformation of reacting groups caused by mismatched base pairs (A.A, A.C) as well as the hybrid rU.dA pair or an unpaired base make the template-directed condensation less effective. The thermal stability of DNA duplexes was assayed before and after the chemical ligation. Among all of the modified duplexes, only the duplex containing 3'-rU in the nick was found to be a substrate of T4 DNA ligase.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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