Filter-binding assay for covalent DNA-protein complexes: adenovirus DNA-terminal protein complex.
A rapid, simple, and quantitative filter-binding assay using glass fiber filters has been developed to detect the convalent adenovirus DNA-terminal protein complex. The assay is unusually sensitive because binding of protein-free DNA generally is less than 0.1%. Binding of the adenovirus complex to filters is mediated by terminal protein. We have found that: (i) the adenovirus complex binds maximally to filters in NaCl at concentrations higher than 0.2 M; (ii) noncovalent complexes between protein-free DNA and adenovirus proteins bind to filters in salt at concentrations lower than 0.4 M but not in concentrations higher than 0.7 M; and (iii) protein-free DNA alone binds to filters in guanidine.hydrochloride at concentrations higher than 0.8 M. By varying the ionic conditions, "all or none" modulation of these interactions can be achieved.
D000256:Adenoviridae; D004279:DNA, Viral / Q000302:isolation & purification; D005374:Filtration; D007700:Kinetics; D009994:Osmolar Concentration; D011485:Protein Binding; D014764:Viral Proteins / Q000302:isolation & purification
D004279:DNA, Viral; D014764:Viral Proteins