Journal Article

Synthesis of novel poly(dG)–poly(dG)–poly(dC) triplex structure by Klenow exo<sup>−</sup> fragment of DNA polymerase I

Alexander Kotlyar, Natalia Borovok, Tatiana Molotsky, Dmitry Klinov, Benjamin Dwir and Eli Kapon

in Nucleic Acids Research

Volume 33, issue 20, pages 6515-6521
Published in print December 2005 | ISSN: 0305-1048
Published online December 2005 | e-ISSN: 1362-4962 | DOI: https://dx.doi.org/10.1093/nar/gki963

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The extension of the G-strand of long (700 bp) poly(dG)–poly(dC) by the Klenow exo fragment of DNA polymerase I yields a complete triplex structure of the H-DNA type. High-performance liquid chromatography analysis demonstrates that the length of the G-strand is doubled during the polymerase synthesis. Fluorescence resonance energy transfer analysis shows that the 5′ ends of the G- and the C-strands, labeled with fluorescein and TAMRA, respectively, are positioned close to each other in the product of the synthesis. Atomic force microscopy morphology imaging shows that the synthesized structures lack single-stranded fragments and have approximately the same length as the parent 700 bp poly(dG)–poly(dC). CD spectrum of the polymer has a large negative peak at 278 nm, which is characteristic of the poly(dG)–poly(dG)–poly(dC) triplex. The polymer is resistant to DNase and interacts much more weakly with ethidium bromide as compared with the double-stranded DNA.

Journal Article.  5213 words.  Illustrated.

Subjects: Molecular and Cell Biology

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