Figure 4.
DRaCALA allows detection of specific interaction of CRP with plasmid carrying the ICAP site. (A) 50 pM individual plasmids with 1×, 3× or 5× wild-type binding sites or 3× mutant-binding sites cloned in series were tested for binding in the presence of 100 nM CRP and 200 µM cAMP. (B) Specificity was determined by competition of binding to 32P-labeled 1× wild-type plasmid with unlabeled PCR products. Competitors used were 1× ICAP, 3× 8:G-C, 3× 10:G-C, 3× 8, 10:G-C. All error bars represent SD of three spots with a representative spot (spot images consolidated to fit graph) shown above each column.