Use of ion exchange chromatography for the study of RecA-DNA interaction |
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| Authors: | M Takahashi P Hagmar |
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| Affiliation: | Institut de Biologie Molécule et Cellulaire du CNRS, Strasbourg, France. |
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| Abstract: | In vitro binding of RecA protein to double-stranded DNA (dsDNA) was studied using ion-exchange liquid chromatography. The method allowed quantification of both free DNA and free protein. The results unambiguously showed a binding stoichiometry of 3 base pairs per RecA monomer. The binding exhibited cooperativity, and the stoichiometry suggested that RecA does not form complexes with two molecules of dsDNA. More than 90% of RecA molecules in the sample were active for DNA binding. |
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