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The DNA‐protein cross: a method for detecting specific DNA‐protein complexes in crude mixtures.
Author(s) -
Patel S.B.,
Cook P.R.
Publication year - 1983
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1002/j.1460-2075.1983.tb01395.x
Subject(s) - biology , dna , repressor , lac repressor , genomic dna , microbiology and biotechnology , agarose , lysogen , biochemistry , lac operon , escherichia coli , gene , gene expression , bacteriophage , plasmid
A method using crude cellular mixtures is described which permits identification of polypeptides and DNA fragments forming specific complexes. Our procedure incorporates elements of both ‘Southern’ and ‘protein’ blotting and combines, in two dimensions, the resolving power of a denaturing protein gel with that of an agarose DNA gel. Conditions for ‘crossing’ have been established using the lambda repressor‐operator system: the specific complex can be detected by crossing total protein from bacteria overproducing the repressor with a mixture of total genomic fragments from a lysogen.