Canonical nucleosides can be utilized by T4 DNA ligase as universal template bases at ligation junctions

Rashada C. Alexander, Ashley K. Johnson, Jeffrey A. Thorpe, Travis Gevedon, Stephen M. Testa

Research output: Contribution to journalArticlepeer-review

28 Scopus citations


T4 DNA ligase catalyzes the template-dependent ligation of DNA. Using T4 DNA ligase under specific experimental conditions, we demonstrate that each of the four canonical nucleosides, centrally located on a template molecule such that they flank the site of ligation, can direct the ligation of nucleic acids regardless of the identity of the terminal nucleosides being covalently joined. This universal templating capability extends to those positions adjacent to the ligation junction. This is the first report, irrespective of the ligation method used or the identity of the template nucleosides (including analogs), which shows that nucleosides can act essentially as universal templates at ligation junctions in vitro. The canonical nucleosides do, however, differ in their ability to template sequence-independent ligations, with thymidine and guanosine being equally effective, yet more effective than adenosine and cytidine. Results indicate that hybridization strength surrounding the ligation junction is an important factor. The implications of this previously undiscovered property of T4 DNA ligase with canonical nucleosides are discussed.

Original languageEnglish
Pages (from-to)3208-3216
Number of pages9
JournalNucleic Acids Research
Issue number12
StatePublished - Jun 15 2003

Bibliographical note

Funding Information:
The authors would like to thank Dr Michael Bell and Dana Baum for helpful discussions. This work was aided by grant 85-001-13-IRG from the American Cancer Society, by an award from the Research Corporation and by the Kentucky Research Challenge Trust Fund.

ASJC Scopus subject areas

  • Genetics


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