Abstract
Studies of the sequence-specific binding of proteins to DNA have so far relied on in vitro experiments using cloned restriction fragments containing the relevent DNA sequences1–6. We have applied the genomic sequencing technique of Church and Gilbert7 to show that the interactions observed in vitro occur in vivo. We use this approach to study the binding of regulatory proteins to the lac operon in vivo and detect changes in the reactivity (inhibition or enhancement) of guanines to methylation by dimethyl sulphate caused by the proximity of proteins to the N-7 atom of these guanines1,5. We can detect the simultaneous binding of the catobolite gene activator protein (CAP) and the Lac represser to their specific recognition sequences, and following induction of the lac operon we observe effects that are related to RNA polymerase binding or RNA elongation. We have successfully used oligonucleotide probes as short as 17 bases to display genomic sequence.
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Nick, H., Gilbert, W. Detection in vivo of protein-DNA interactions within the lac operon of Escherichia coli. Nature 313, 795–798 (1985). https://doi.org/10.1038/313795a0
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DOI: https://doi.org/10.1038/313795a0
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