Wold W S, Green M, Mackey J K, Martin J D, Padgett B L, Walker D L
J Virol. 1980 Mar;33(3):1225-8. doi: 10.1128/JVI.33.3.1225-1228.1980.
The physical state of the JC virus (JCV) genome was studied in two clonal cell lines (clones 2 and 7) derived from a tissue culture cell line (HJC-15) established from a hamster brain tumor induced by JCV. Saturation-hybridization and reassociation kinetic analyses, using in vitro (32)P-labeled JCV DNA, indicated that clone 7 and 2 cells contain 9 to 10 and 4 to 5 copies per cell, respectively, of all or most of the viral genome. Both cell DNAs were analyzed by using the Southern blotting procedure with three restriction endonucleases: XhoI, which does not cleave JCV DNA; EcoRI, which cleaves once; and HindIII, which cleaves three times. With each DNA, a variety of JCV-specific DNA fragments were detected. The following conclusions are possible: (i) JCV DNA is integrated into cell DNA in both clonal lines; (ii) both clonal lines contain multiple copies of the viral genome integrated in a tandem head-to-tail orientation; (iii) neither clonal line contains detectable free-form I, II, or III JCV DNA; (iv) each clonal line contains multiple independent sites of JCV DNA integration; and (v) most or all of the sites of integration on the cellular or the viral genome, or both, are different in clone 7 DNA than in clone 2 DNA. Thus, although both clone 7 and clone 2 cells were established from the HJC-15 tumor cell line, they differ in the copy number and integration pattern of JCV DNA.
在源自由JC病毒诱导的仓鼠脑肿瘤建立的组织培养细胞系（HJC - 15）的两个克隆细胞系（克隆2和克隆7）中研究了JC病毒（JCV）基因组的物理状态。使用体外（32）P标记的JCV DNA进行饱和杂交和复性动力学分析表明，克隆7和克隆2细胞中每个细胞分别含有病毒基因组全部或大部分的9至10个拷贝和4至5个拷贝。使用三种限制性内切酶通过Southern印迹法分析了两种细胞DNA：XhoI，它不切割JCV DNA；EcoRI，它切割一次；HindIII，它切割三次。对于每种DNA，检测到多种JCV特异性DNA片段。可能得出以下结论：（i）JCV DNA在两个克隆系中均整合到细胞DNA中；（ii）两个克隆系均含有以头尾串联方向整合的病毒基因组的多个拷贝；（iii）两个克隆系均不含有可检测到的游离形式的I、II或III型JCV DNA；（iv）每个克隆系均含有JCV DNA整合的多个独立位点；（v）克隆7 DNA与克隆2 DNA相比，细胞基因组或病毒基因组上或两者上的大多数或所有整合位点均不同。因此，尽管克隆7和克隆2细胞均源自HJC - 15肿瘤细胞系，但它们在JCV DNA的拷贝数和整合模式上存在差异。
