Method to generate Holliday junction recombination intermediates via RecA-mediated four-strand exchange

Anal Biochem. 2023 Oct 14;682:115347. doi: 10.1016/j.ab.2023.115347. Online ahead of print.ABSTRACTDNA molecules that contain single Holliday junctions have served as model substrates to investigate the pathway in which homologous recombination intermediates are processed. However, the preparation of DNA containing Holliday junctions in high yield remains a challenge. In this work, we used a nicking endonuclease to generate gapped DNA, from which α-structured DNA or figure-8 DNA were created via RecA-mediated reactions. The resulting DNA molecules were found to serve as good substrates for Holliday junction resolvases. The simplified method negates the requirement for radioactive labelling of DNA, making the generation of Holliday junction DNA more accessible to non-experts.PMID:37821038 | DOI:10.1016/j.ab.2023.115347

Source: Analytical Biochemistry - October 11, 2023 Category: Biochemistry Authors: Han Ngoc Ho Stephen C West Source Type: research