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Two‐dimensional gel selection of protein binding sites on DNA
Author(s) -
Boffini Andrea,
Prentki Pierre
Publication year - 1993
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150140146
Subject(s) - dna , dna binding protein , electrophoretic mobility shift assay , cloning (programming) , escherichia coli , gel electrophoresis , electrophoresis , genome , biology , selection (genetic algorithm) , chemistry , computational biology , microbiology and biotechnology , genetics , transcription factor , gene , computer science , artificial intelligence , programming language
We have developed a gel electrophoretic approach for visualizing and cloning protein binding sites from complete genomes. This system consists of a simple two‐dimensional band shift, in which protein‐DNA complexes are retarded in the first dimension, performed at low temperature, and disrupted in the second dimesion, performed at high temperature. We present here results obtained with the integration host factor (IHF) and cAMP receptor protein (CRP) proteins of Escherichia coli , and discuss some of the important methodological aspects of the technique.