Single-Stranded DNA Curtains for Studying Homologous Recombination.
Methods Enzymol
; 582: 193-219, 2017.
Article
em En
| MEDLINE
| ID: mdl-28062035
ABSTRACT
Homologous recombination is an important pathway involved in the repair of double-stranded DNA breaks. Genetic studies form the foundation of our knowledge on homologous recombination. Significant progress has also been made toward understanding the biochemical and biophysical properties of the proteins, complexes, and reaction intermediates involved in this essential DNA repair pathway. However, heterogeneous or transient recombination intermediates remain extremely difficult to assess through traditional ensemble methods, leaving an incomplete mechanistic picture of many steps that take place during homologous recombination. To help overcome some of these limitations, we have established DNA curtain methodologies as an experimental platform for studying homologous DNA recombination in real-time at the single-molecule level. Here, we present a detailed overview describing the preparation and use of single-stranded DNA curtains in applications related to the study of homologous DNA recombination with emphasis on recent work related to the study of the eukaryotic recombinase Rad51.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Rad51 Recombinase
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Recombinação Homóloga
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Imagem Individual de Molécula
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Microscopia de Fluorescência
Idioma:
En
Revista:
Methods Enzymol
Ano de publicação:
2017
Tipo de documento:
Article
País de afiliação:
Estados Unidos