Green M R, Chinnadurai G, Mackey J K, Green M
Cell. 1976 Mar;7(3):419-28. doi: 10.1016/0092-8674(76)90172-0.
We have analyzed the viral DNA sequences persisting in HE C19, a clonal line of hamster embryo cells transformed in vitro by human adenovirus 12 (Ad 12), a member of highly oncogenic group A. Saturation-hybridization of in vitro labeled Ad 12 DNA (2-4 x 10(8) cpm/mug) with HE C19 DNA showed that 93-100% of viral DNA sequences are present in the transformed cell. Reassociation of Ad 12 32P-DNA in the presence of HE C19 DNA detected 8 viral genome equivalents per diploid quantity of cell DNA. REannealing each of the six Ad 12 EcoRl restriction DNA fragments with HE C19 DNA indicated that the sequences of each fragment are present in near equimolar quantities, 8-10.5 copies per cell. HE C19 DNA "networks" were prepared and shown to contain the same viral DNA sequences and in the same amounts as present in unfractionated transformed cell DNA; these data provide the first direct evidence that human adenovirus genes are covalently linked to cellular DNA. The presence of most, if not all, of the viral genes was demonstrated also in the Ad 12 hamster tumor cell line, HT-2. The pattern of integrated viral genes in these nonpermissive hamster cells transformed by Ad 12 is different from that described for cells transformed by group C and B human adenoviruses and for SV40.
我们分析了存在于HE C19中的病毒DNA序列,HE C19是一种仓鼠胚胎细胞的克隆系,由人腺病毒12(Ad 12)在体外转化而来,Ad 12是高致癌性A组的成员。用体外标记的Ad 12 DNA(2 - 4×10⁸ cpm/μg)与HE C19 DNA进行饱和杂交,结果显示93 - 100%的病毒DNA序列存在于转化细胞中。在HE C19 DNA存在的情况下,Ad 12 ³²P - DNA的复性检测到每二倍体细胞DNA量中有8个病毒基因组当量。将六个Ad 12 EcoRl限制性DNA片段中的每一个与HE C19 DNA进行再退火,表明每个片段的序列以近等摩尔量存在,每个细胞有8 - 10.5个拷贝。制备了HE C19 DNA“网络”,并显示其含有与未分级的转化细胞DNA中相同的病毒DNA序列,且数量相同;这些数据提供了首个直接证据,证明人腺病毒基因与细胞DNA共价连接。在Ad 12仓鼠肿瘤细胞系HT - 2中也证实了大部分(如果不是全部)病毒基因的存在。在这些由Ad 12转化的非允许性仓鼠细胞中,整合病毒基因的模式与由C组和B组人腺病毒以及SV40转化的细胞中所描述的模式不同。