Fluorescently labeled circular DNA molecules for DNA topology and topoisomerases

Authors

Maxwell Gu, Biomolecular Sciences Institute and Department of Chemistry and Biochemistry, Florida International UniversityFollow
Andrea Berrido, Biomolecular Sciences Institute and Department of Chemistry and Biochemistry, Florida International UniversityFollow
Walter Gonzalez, Biomolecular Sciences Institute and Department of Chemistry and Biochemistry, Florida International UniversityFollow
Jaroslava Miksovska, Biomolecular Sciences Institute and Department of Chemistry and Biochemistry, Florida International UniversityFollow
Jeremy W. Chambers, Biomolecular Sciences Institute and Department of Chemistry and Biochemistry, Florida International UniversityFollow
Fenfei Leng, Biomolecular Sciences Institute and Department of Chemistry and Biochemistry, Florida International UniversityFollow

Date of this Version

10-31-2016

Document Type

Article

Rights

by

Abstract

DNA topology plays essential roles in several fundamental biological processes, such as DNA replication, recombination, and transcription. Typically agarose gel electrophoresis is employed to study DNA topology. Since gel electrophoresis is time-consuming and labor intensive, it is desirable to develop other methods, such as fluorescence-based methods, for such studies. In this paper we report the synthesis of a type of unique fluorescence-labeled DNA molecules that can be used to study DNA topology and topoisomerases by fluorescence resonance energy transfer (FRET). Specifically, we inserted an 82 nt. synthetic DNA oligomer FL905 carrying a 42 nt. AT sequence with fluorescein and dabcyl labels into a gapped DNA molecule to generate relaxed and supercoiled pAB1_FL905. Since the fluorescence intensity of pAB1_FL905 is dependent on its supercoiling status, pAB1_FL905 is a powerful tool to study DNA topology and topoisomerases by FRET. pAB1_FL905 can also be developed into rapid and efficient high-throughput screening assays to identify inhibitors that target various DNA topoisomerases.

DOI

10.1038/srep36006

Identifier

FIDC001613

Comments

Originally published in Scientific Reports.

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 License.

Recommended Citation

Gu, M. et al. Fluorescently labeled circular DNA molecules for DNA topology and topoisomerases. Sci. Rep. 6, 36006; doi: 10.1038/srep36006 (2016).