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Bands on the run

Nature Methods volume 2, page 556 (2005)Cite this article

Scientists have been trying to understand the nature and importance of protein-DNA interactions since the early 1960s. Such research started with studies of the association of RNA polymerase with DNA, but soon expanded to encompass the characterization of two newly-discovered but equally intriguing DNA-binding proteins: the Lac repressor1 and the phage λ repressor2.

One of the most important early techniques for analyzing these complexes arose from the discovery that certain membrane filters will retain DNA-protein complexes, but not free DNA3. By quantifying the retention of radiolabeled DNA fragments mixed with varying amounts of a protein of interest, it became possible to determine with reasonable accuracy the stoichiometry and binding affinity of a protein for a given sequence, and the technique was soon being successfully applied for the analysis of RNA polymerase3 and the Lac repressor4. But even though filter binding became relatively popular, it remained impractical for the characterization of less stable complexes, and could only be used to demonstrate the existence of DNA-protein complexes without necessary revealing all of the participating components.

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Authors
  1. Michael Eisenstein
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Eisenstein, M. Bands on the run. Nat Methods 2, 556 (2005). https://doi.org/10.1038/nmeth0705-556

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