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Nucleic Acids Research, 1991, Vol. 19, No. 7 1369-1374
© 1991

MOLECULAR BIOLOGY

Identification of protein binding sites in genomic DNA by two-dimensional gel electrophoresis

Andrea Boffini and Pierre Prentki*

Department of Molecular Biology, University of Geneva 1211 Geneva 4, Switzerland

* To whom correspondence should be addressed

Received February 12, 1991. Revised March 14, 1991. Accepted March 14, 1991.

We describe a simple two-dimensional electrophoresis procedure to identify the recognition sites of DNA-binding proteins within large DNA molecules. Using this approach, we have mapped E.coli IHF (Integration Host Factor) binding sites within phage Lambda (48 kb) and phage Mu (39 kb) DNA. We are also able to visualize IHF binding sites in E.coli chromosomal DNA (4,700 kb). We present an extension of this technique using direct amplification by PCR of the isolated restriction fragments, which should permit the cloning of a collection of recognition sequences for DNA binding proteins in complex genomes.


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