Filter-binding assay for covalent DNA-protein complexes: adenovirus DNA-terminal protein complex.
Filter-binding assay for covalent DNA-protein complexes: adenovirus DNA-terminal protein complex.
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DOI：
10.1073/pnas.75.11.5291
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发表时间：
1978-11
影响因子：
11.1
通讯作者：
D. Coombs;G. D. Pearson
中科院分区：
文献类型：
--
作者：
D. Coombs;G. D. Pearson
一种快速、简便、定量的玻璃纤维滤膜结合试验已被开发用于检测共价腺病毒DNA-末端蛋白复合物。该测定法异常灵敏，因为无蛋白DNA的结合通常小于0.1%。腺病毒复合物与滤器的结合由末端蛋白介导。我们发现：（i）在浓度高于0.2M的NaCl中，腺病毒复合物最大程度地结合到滤膜上;（ii）在浓度低于0.4M但不高于0.7M的盐中，无蛋白DNA和腺病毒蛋白之间的非共价复合物结合到滤膜上;和（iii）在浓度高于0.8M的盐酸胍中，无蛋白DNA单独结合到滤膜上。通过改变离子条件，可以实现这些相互作用的“全部或全部不”调节。
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.