Biochemical Analysis of D-Loop Extension and DNA Strand Displacement Synthesis
Homologous recombination is a conserved genome maintenance pathway through which DNA double-strand breaks are eliminated and perturbed DNA replication forks and eroded telomeres are restored. The central step in homologous recombination is homology-dependent pairing between a single-stranded DNA tail with an intact duplex molecule to generate a displacement-loop (D-loop), followed by DNA synthesis within the D-loop platform. This chapter describes biochemical assays for (1) D-loop formation and DNA synthesis within the D-loop and (2) DNA strand displacement synthesis to test the role of DNA helicases (e.g., Pif1) in repair DNA synthesis. These mechanistic assays are valuable for elucidating the molecular details of HR.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2021 |
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| Erschienen: |
2021 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:2153 |
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| Enthalten in: |
Methods in molecular biology (Clifton, N.J.) - 2153(2021) vom: 25., Seite 87-99 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Kwon, Youngho [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 17.03.2021 Date Revised 17.03.2021 published: Print Citation Status MEDLINE |
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| doi: |
10.1007/978-1-0716-0644-5_7 |
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| funding: |
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NLM314141731 |
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| 520 | |a Homologous recombination is a conserved genome maintenance pathway through which DNA double-strand breaks are eliminated and perturbed DNA replication forks and eroded telomeres are restored. The central step in homologous recombination is homology-dependent pairing between a single-stranded DNA tail with an intact duplex molecule to generate a displacement-loop (D-loop), followed by DNA synthesis within the D-loop platform. This chapter describes biochemical assays for (1) D-loop formation and DNA synthesis within the D-loop and (2) DNA strand displacement synthesis to test the role of DNA helicases (e.g., Pif1) in repair DNA synthesis. These mechanistic assays are valuable for elucidating the molecular details of HR | ||
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