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

ABSTRACT: 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).

SUBMITTER: Yao C

PROVIDER: S-EPMC8406033 | biostudies-literature | 2021 Sep

REPOSITORIES: biostudies-literature

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

Yao Chunyan C Shao Xiaolong X Li Jingwei J Deng Xin X

STAR protocols 20210825 3

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 electroporati ...[more]

PMID: 34485942

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