Fujinaga K, Sawada Y, Uemizu Y
Gan. 1979 Apr;70(2):239-43.
The viral DNA sequences in cells transformed by adenovirus type-12 (Ad12) DNA fragments were investigated by spot hybridization, a detection by autoradiography of nucleic acid hybrids formed between cell DNAs spotted on a membrane filter and various nick-translated Ad12 DNA fragments. In CY1 cell line, a rat cell line transformed by the EcoRI-C fragment (left hand 16%), all of the HindIII fragments included in the EcoRI-C fragment are shown to be present. In GY1 cell line, a rat cell line transformed by the HindIII-G fragment (left hand 7%), both of the BpaI-H and a part of the BpaI-J, two components consisting of the HindIII-G fragment, were found. A dominant presence of the Ad12 BpaI-H fragment (left hand end 4.5%) of the Ad12 DNA molecule, approximately 60% of the Ad12 transforming DNA sequences (Ad12 HindIII-G), was also shown in GY1. Spot hybridization used in the experiment will be of general use for detecting viral nucleic acid sequences in cells and provides a simple and useful screening method for investigating viral etiology of tumors and transformed cells.
通过斑点杂交研究了腺病毒12型（Ad12）DNA片段转化细胞中的病毒DNA序列，斑点杂交是通过放射自显影检测膜滤纸上点样的细胞DNA与各种切口平移的Ad12 DNA片段之间形成的核酸杂交体。在CY1细胞系（一种由EcoRI-C片段（左手端16%）转化的大鼠细胞系）中，显示EcoRI-C片段中包含的所有HindIII片段均存在。在GY1细胞系（一种由HindIII-G片段（左手端7%）转化的大鼠细胞系）中，发现了HindIII-G片段的两个组成部分BpaI-H和部分BpaI-J。在GY1中还显示了Ad12 DNA分子的Ad12 BpaI-H片段（左手端4.5%）占主导地位，约占Ad12转化DNA序列（Ad12 HindIII-G）的60%。实验中使用的斑点杂交将普遍用于检测细胞中的病毒核酸序列，并为研究肿瘤和转化细胞的病毒病因提供一种简单而有用的筛选方法。
