GLUTATHIONE-S-TRANSFERASE FUSION PROTEINS AS AN AFFINITY REAGENT FOR RAPID ISOLATION OF SPECIFIC SEQUENCE DIRECTLY FROM GENOMIC DNA

We describe a DNA binding assay for isolation of specific sequence(s) recognized by protein of interest directly from genomic or cosmid DNA. In our assay, the protein is fused to the glutathione-S-transferase a nd bound to glutathione-Sepharose beads. Then the immobilized fusion p rotein can be used to search for DNA fragment(s) that interact specifi cally with the protein of interest. As an example of such an approach, we identified and cloned a few prokaryotic oriC regions using the ini tiator DnaA protein fused to the glutathione-S-transferase.