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Fig. 1 | BMC Research Notes

Fig. 1

From: An enChIP system for the analysis of bacterial genome functions

Fig. 1

An enChIP system for biochemical analysis of bacterial genome functions. a Schematic of in-cell enChIP. Engineered DNA-binding molecules such as CRISPR can be used for locus tagging. The isolated materials can be subjected to biochemical analyses such as MS or NGS for identification of interacting molecules. b Inducible expression of 3xFLAG-dCas9. DH5α cells were transformed with 3xFLAG-dCas9/p-bacteria. The bacterial cells were incubated in the presence of doxycycline (Dox, 2 µM) for 4 h and subjected to immunoblot analysis with anti-FLAG M2 Ab. Coomassie brilliant blue (CBB) staining is shown as a protein loading control. c Positions of a gRNA and primer sets targeting the lacZ gene. The positions of primer sets are shown in red. The target sequence of the gRNA is shown, and the protospacer adjacent motif (PAM) sequence is underlined. d Isolation of the lacZ locus by enChIP. Real-time PCR analysis of chromatin complexes isolated by enChIP is shown. Irrelevant loci (crp and lacI) were analyzed as negative controls

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