Yang Jihong, Hearing Patrick
Methods Mol Med. 2007;131:113-21. doi: 10.1007/978-1-59745-277-9_9.
The encapsidation of adenovirus DNA into virus particles depends on cis-acting sequences located at the left end of the viral genome. Repeated DNA sequences in the packaging domain contribute to viral DNA encapsidation, and several viral proteins bind to these repeats when analyzed using in vitro DNA-protein-binding assays. This chapter describes a chromatin immunoprecipitation approach to study the binding of viral proteins to packaging sequences in vivo. The technique is easily adaptable to study the interaction of any viral or cellular protein to Ad DNA or to cellular genomic DNA sequences. The assay permits accurate quantification over a wide range of DNA concentrations. The use of formaldehyde cross-linking to stabilize DNA-protein and protein-protein complexes formed in vivo allows the identification of macromolecular complexes found in living cells.
腺病毒DNA包装进病毒颗粒取决于位于病毒基因组左端的顺式作用序列。包装结构域中的重复DNA序列有助于病毒DNA的包装，并且当使用体外DNA-蛋白质结合试验进行分析时，几种病毒蛋白会与这些重复序列结合。本章描述了一种染色质免疫沉淀方法，用于研究病毒蛋白在体内与包装序列的结合。该技术很容易适用于研究任何病毒或细胞蛋白与腺病毒DNA或细胞基因组DNA序列的相互作用。该试验允许在很宽的DNA浓度范围内进行准确的定量。使用甲醛交联来稳定体内形成的DNA-蛋白质和蛋白质-蛋白质复合物，可以鉴定活细胞中发现的大分子复合物。
