Protocol for Purification of Human ZGRF1 and Its Regulatory Function on RAD51-Mediated D-Loop Formation

Human ZGRF1 is one of the many helicases that play important roles in the repair of damaged chromosomes by homologous recombination. However, the lack of biochemical characterization of the ZGRF1 protein has hindered the understanding of its function. We have developed a facile protocol to express h...

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Detalles Bibliográficos
Autores principales: Lisby, Michael, Xue, Xiaoyu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7518512/
https://www.ncbi.nlm.nih.gov/pubmed/32984852
http://dx.doi.org/10.1016/j.xpro.2020.100099
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author Lisby, Michael
Xue, Xiaoyu
author_facet Lisby, Michael
Xue, Xiaoyu
author_sort Lisby, Michael
collection PubMed
description Human ZGRF1 is one of the many helicases that play important roles in the repair of damaged chromosomes by homologous recombination. However, the lack of biochemical characterization of the ZGRF1 protein has hindered the understanding of its function. We have developed a facile protocol to express human ZGRF1 in insect cells and for its purification. We have also tuned biochemical assays by lowering the protein and DNA concentrations to analyze its functions in RAD51-mediated D-loop formation and dissociation. For complete details on the use and execution of this protocol, please refer to Brannvoll et al. (2020).
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spelling pubmed-75185122020-09-25 Protocol for Purification of Human ZGRF1 and Its Regulatory Function on RAD51-Mediated D-Loop Formation Lisby, Michael Xue, Xiaoyu STAR Protoc Protocol Human ZGRF1 is one of the many helicases that play important roles in the repair of damaged chromosomes by homologous recombination. However, the lack of biochemical characterization of the ZGRF1 protein has hindered the understanding of its function. We have developed a facile protocol to express human ZGRF1 in insect cells and for its purification. We have also tuned biochemical assays by lowering the protein and DNA concentrations to analyze its functions in RAD51-mediated D-loop formation and dissociation. For complete details on the use and execution of this protocol, please refer to Brannvoll et al. (2020). Elsevier 2020-09-01 /pmc/articles/PMC7518512/ /pubmed/32984852 http://dx.doi.org/10.1016/j.xpro.2020.100099 Text en © 2020 The Author(s) http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Lisby, Michael
Xue, Xiaoyu
Protocol for Purification of Human ZGRF1 and Its Regulatory Function on RAD51-Mediated D-Loop Formation
title Protocol for Purification of Human ZGRF1 and Its Regulatory Function on RAD51-Mediated D-Loop Formation
title_full Protocol for Purification of Human ZGRF1 and Its Regulatory Function on RAD51-Mediated D-Loop Formation
title_fullStr Protocol for Purification of Human ZGRF1 and Its Regulatory Function on RAD51-Mediated D-Loop Formation
title_full_unstemmed Protocol for Purification of Human ZGRF1 and Its Regulatory Function on RAD51-Mediated D-Loop Formation
title_short Protocol for Purification of Human ZGRF1 and Its Regulatory Function on RAD51-Mediated D-Loop Formation
title_sort protocol for purification of human zgrf1 and its regulatory function on rad51-mediated d-loop formation
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7518512/
https://www.ncbi.nlm.nih.gov/pubmed/32984852
http://dx.doi.org/10.1016/j.xpro.2020.100099
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