Title

2',2'-Ligation Demonstrates the Thermal Dependence of DNA-directed Positional Control

Abstract

An oligodeoxynucleotide (ODN) containing a 2'-aminoalkylmercaptouridine was ligated chemically with another ODN containing a 2'-carboxyalkylmercaptouridine to produce a 2',2'-amide linked conjugate. Templation by a DNA hairpin resulted in ligation yields of >95%. at room temperature, similar yields were obtained for amide ligations between pendent nucleotides located adjacent to each other or separated by three nucleotides. However, at 4 °C, ligation was vastly favored for the case where the pendent amine and carboxylate were attached to adjacent residues (i.e., at the 3'- and 5'-terminus of the two ODNs). No reaction was observed to occur either at 4 °C or at room temperature between pendent nucleotides separated by nine nucleotides. Denaturing gel electrophoresis, MALDI-TOF MS, and full nuclease digestion coupled with LC-MS were used to characterize the reaction products.

Department(s)

Chemistry

International Standard Serial Number (ISSN)

404020

Document Type

Article - Journal

Document Version

Citation

File Type

text

Language(s)

English

Rights

© 2008 Elsevier, All rights reserved.


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