Explorative Studies On Quantitative Determination Of Protein-DNA Binding Dissociation Constant Using GFP As Label

Understanding gene regulatory mechanism andbiological processes based on protein-nucleic acidinteractions requires biochemical analysis of the bindingreactions. In this study, a rapid quantitative DNA bindingassay was developed using the fusion protein of a DNA -binding protein (i.e. EREBP2) and green fluorescent protein(GFP). This hydrolysis activity was used quantify theamount of EREBP2-GFP bound to an immobilized biotin-labeled target sequence. The preliminary results of this studyindicated that quantitative determination of protein-DNAbinding dissociation constant (Kd) was feasible through...