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Optimized protocols for ChIP-seq and deletion mutant construction in Pseudomonas syringae

Chromatin immunoprecipitation sequencing (ChIP-seq) is an efficient technique to identify the binding sites of transcription factors (TFs) in both eukaryotes and prokaryotes. However, its application in bacteria is very heterogeneous. In this protocol, we optimized the methods of ChIP-seq that can b...

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Detalles Bibliográficos
Autores principales: Yao, Chunyan, Shao, Xiaolong, Li, Jingwei, Deng, Xin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8406033/
https://www.ncbi.nlm.nih.gov/pubmed/34485942
http://dx.doi.org/10.1016/j.xpro.2021.100776
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author Yao, Chunyan
Shao, Xiaolong
Li, Jingwei
Deng, Xin
author_facet Yao, Chunyan
Shao, Xiaolong
Li, Jingwei
Deng, Xin
author_sort Yao, Chunyan
collection PubMed
description Chromatin immunoprecipitation sequencing (ChIP-seq) is an efficient technique to identify the binding sites of transcription factors (TFs) in both eukaryotes and prokaryotes. However, its application in bacteria is very heterogeneous. In this protocol, we optimized the methods of ChIP-seq that can be widely applied to plant pathogens. We used homologous recombination to construct pK18mobsacB-Psph plasmid instead of restriction site ligation and replaced transconjugation with electroporation transformation in Pseudomonas syringae deletion mutant construction, which is more efficient and faster than previous methods. For complete details on the use and execution of this protocol, please refer to Shao et al. (2021).
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spelling pubmed-84060332021-09-02 Optimized protocols for ChIP-seq and deletion mutant construction in Pseudomonas syringae Yao, Chunyan Shao, Xiaolong Li, Jingwei Deng, Xin STAR Protoc Protocol Chromatin immunoprecipitation sequencing (ChIP-seq) is an efficient technique to identify the binding sites of transcription factors (TFs) in both eukaryotes and prokaryotes. However, its application in bacteria is very heterogeneous. In this protocol, we optimized the methods of ChIP-seq that can be widely applied to plant pathogens. We used homologous recombination to construct pK18mobsacB-Psph plasmid instead of restriction site ligation and replaced transconjugation with electroporation transformation in Pseudomonas syringae deletion mutant construction, which is more efficient and faster than previous methods. For complete details on the use and execution of this protocol, please refer to Shao et al. (2021). Elsevier 2021-08-25 /pmc/articles/PMC8406033/ /pubmed/34485942 http://dx.doi.org/10.1016/j.xpro.2021.100776 Text en © 2021 The Author(s) https://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
Yao, Chunyan
Shao, Xiaolong
Li, Jingwei
Deng, Xin
Optimized protocols for ChIP-seq and deletion mutant construction in Pseudomonas syringae
title Optimized protocols for ChIP-seq and deletion mutant construction in Pseudomonas syringae
title_full Optimized protocols for ChIP-seq and deletion mutant construction in Pseudomonas syringae
title_fullStr Optimized protocols for ChIP-seq and deletion mutant construction in Pseudomonas syringae
title_full_unstemmed Optimized protocols for ChIP-seq and deletion mutant construction in Pseudomonas syringae
title_short Optimized protocols for ChIP-seq and deletion mutant construction in Pseudomonas syringae
title_sort optimized protocols for chip-seq and deletion mutant construction in pseudomonas syringae
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8406033/
https://www.ncbi.nlm.nih.gov/pubmed/34485942
http://dx.doi.org/10.1016/j.xpro.2021.100776
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