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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...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2021
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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). |
format | Online Article Text |
id | pubmed-8406033 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
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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