Chapter title |
Synthesis of Hemicatenanes for the Study of Type IA Topoisomerases
|
---|---|
Chapter number | 11 |
Book title |
DNA Topoisomerases
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7459-7_11 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7458-0, 978-1-4939-7459-7
|
Authors |
Shun-Hsiao Lee, Tao-shih Hsieh, Grace Ee-Lu Siaw, Lee, Shun-Hsiao, Hsieh, Tao-shih, Siaw, Grace Ee-Lu |
Abstract |
Hemicatenane is a structure that forms when two DNA duplexes are physically linked through a single-stranded crossover. It is proposed to be an intermediate resulting from double Holliday junction (dHJ) dissolution, repair of replication stalled forks and late stage replication. Our previous study has shown that hemicatenane can be synthesized and dissolved in vitro by hyperthermophilic type IA topoisomerases. Here we present the protocol of hemicatenane synthesis and its structure detection by 2D agarose gel electrophoresis. The generated product can be used as a substrate to study the biochemical mechanism of hemicatenane processing reactions. |
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