ChIP-seq study of sigma factors in Pseudomonas aeruginosa
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ABSTRACT: Sigma factors are master regulators of bacterial transcription which direct gene expression of specific subsets of genes. In particular, alternative sigma factors are well-known to be key players of bacterial adaptation to changing environments. To elucidate the regulatory network of sigma factors in P. aeruginosa, an integrative approach including ChIP-seq of 11 polyhistidine-tag sigma factors was performed to define the primary regulon of each sigma factor. Sigma factor genes were fused to a polyhistidine-tag and provided in trans. Under optimal conditions regarding sigma factor activity and induction of sigma factor expression, DNA-sigma factor interactions were conserved by formaldehyde treatment. Upon DNA fragmentation by sonication, the complexes were specifically immunoprecipitated by polyclonal anti-6X His-tag antibodies and the purified DNA was analyzed by Illumina sequencing. DNA enrichment to a control strain was calculated and used for peak calling within promoter region (-500,+100) to identify directly regulated genes/operons.
ORGANISM(S): Pseudomonas aeruginosa
SUBMITTER: Denitsa Eckweiler
PROVIDER: E-GEOD-54997 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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