Separation of carcinogen-damaged DNA fragments from undamaged DNA.
Separation of carcinogen-damaged DNA fragments from undamaged DNA.
复制标题
将致癌物质损伤的 DNA 片段与未损伤的 DNA 分离。
DOI：
10.1093/nar/19.24.6955
复制
发表时间：
1991
影响因子：
14.9
通讯作者：
Gentile,C
中科院分区：
文献类型：
--
作者：
Strickland,PT;Gentile,C
Immunological methods employing antibodies specific for carcinogen-damaged DNA have been used to detect DNA damage in human and animal cells (1, 2). These antibodies also provide a potential approach to examining the distribution of carcinogen damage within the genome. We have developed a separation procedure based on immunoprecipitation to separate carcinogen-damaged DNA fragments from undamaged DNA. The procedure involves (i) digestion of carcinogen-damaged DNA with restriction enzyme,(ii) precipitation of complexes between damage-specific monoclonal antibodies and carcinogen-damaged DNA fragments using anti-mouse secondary antibody under conditions where undamaged DNA fragments do not precipitate, and (iii) electrophoretic fractionation of damaged-DNA fragments on agarose gels. Fragments can subsequently be identified by Southern transfer to nitrocellulose and gene-specific DNA hybridization. The procedure has been demonstrated using a monoclonal antibody specific for UV-induced DNA photoproducts to separate damaged and undamaged fragments of UV-irradiated DNA (Figure 1) in a dose-dependent manner. This approach should also be useful in separating DNA damaged by chemical carcinogens when antibodies specific for chemical carcinogen-modified DNA are used.
