Chromatin immunoprecipitation DNA-sequencing (ChIP-seq) for RNA-polymerases encoded by P23-45 bacteriophage

By doing ChIP-seq with antibodies against two P23-45 RNA polymerases gp64 and gp96 we showed that both RNA polymerases interact with the phage genome at the early stage of infection. Gp96 interacts with pre-early P23-45 genes, while gp64 interacts with pre-early, early and some middle-stage genes. Overall design: Thermus thermophilus HB8 cells were infected with P23-45 bacteriophage at a high multiplicity of infection. Next, cells were collected at 5, 20, and 40 minutes post infection with the phage. Chromatin immunoprecipitation followed by DNA-sequencing (ChIP-seq) was performed using antibodies against P23-45 RNA polymerases gp64 and gp96 (first 1540 amino acid residues). Samples without addition of antibodies were used as controls for input DNA.