Fig. 2: E. coli O157:H7 CMoRE specifically degrades 5hmC- or 5ghmC-modified DNA.
From: A bacterial defense system targeting modified cytosine of phage genomic DNA
a CMoRE degrades the genomic DNA of T-even phages. b Quantitative PCR analysis of T4 phage DNA in the presence or absence of the CMoRE system. At each time post infection, phage-specific primers are used for the measurement, with host 16S rRNA serving as an internal normalization control. Bar graphs represent the mean of three biological replicates. c Normalized next-generation sequencing reads mapped to the T4 phage and E. coli genomes. DNA samples for sequencing are harvested 8 min post-infection. d Endonuclease activity of CMoRE against PCR-amplified products with various cytosine modifications. e Electrophoretic mobility shift assay of CMoRE binding to PCR-amplified DNA containing unmodified cytosine, 5mC, 5hmC, or 5ghmC. Increasing concentrations of CMoRE proteins (0, 0.1, 0.2, 0.5, 1, 2, or 5 μM) are incubated with 0.1 μM DNA. f Schematic diagram of library construction and sequencing workflow for CMoRE-digested DNA fragments. g Sequence logo representation of the CMoRE cleavage sites. h Endonuclease activity of CMoRE against synthesized modified DNA substrates. Source data are provided with this paper.
