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.
Recommended Citation
Y. Liu et al., "2',2'-Ligation Demonstrates the Thermal Dependence of DNA-directed Positional Control," Tetrahedron, vol. 64, no. 36, pp. 8417 - 8422, Elsevier, Sep 2008.
The definitive version is available at https://doi.org/10.1016/j.tet.2008.05.046
Department(s)
Chemistry
Keywords and Phrases
Nucleic Acids; Positional Synthesis; Reaction Regioselectivity; Templated Chemical Ligation; Thermal Control
International Standard Serial Number (ISSN)
0040-4020
Document Type
Article - Journal
Document Version
Citation
File Type
text
Language(s)
English
Rights
© 2008 Elsevier, All rights reserved.
Publication Date
01 Sep 2008
