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Figure 1 | BMC Molecular Biology

Figure 1

From: Efficient isolation of specific genomic regions retaining molecular interactions by the iChIP system using recombinant exogenous DNA-binding proteins

Figure 1

Scheme of iChIP using r3xFNLDD-D. 3xFNLDD-D consisting of 3xFLAG-tag, a nuclear localization signal (NLS), the DNA-binding domain (DB) plus the dimerization domain of the LexA protein, and Dock-tag, is expressed and purified. The recognition sequences of the LexA protein (LexA BE) are inserted into a genomic region of interest, usually by homologous recombination, in the cell to be analyzed. The resultant cell is stimulated and crosslinked with formaldehyde or other crosslinkers, if necessary. The cell is lysed, and the genomic DNA is fragmented. The target genomic region is affinity purified with r3xFNLDD-D conjugated with anti-FLAG antibody (Ab). After reverse crosslinking, if necessary, purification of the chromatin components (DNA, RNA, proteins, other molecules) allows their identification and characterization.

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