Van der Eb A J, Houweling A
Gene. 1977;2(3-4):133-46. doi: 10.1016/0378-1119(77)90013-0.
Five clones of rat kidney cells transformed by a small restriction endonuclease fragment of adenovirus 5 (Ad5) DNA (fragment HsuI G, which represents the left terminal 7% of the adenovirus genome) were analyzed with respect to the viral DNA sequences present in the cellular DNAs. In these analyses, the kinetics of renaturation of 32P-labeled specific fragments of Ad5 DNA was measured in the presence of a large amount of DNA extracted either from each of the transformed cell lines or from untransformed cells. The fragments were produced by digestion of 32P-labeled adenovirus 5 DNA with endo R.HsuI, or by digestion of 32P-labeled fragment HsuI G of adeno 5 DNA with endo R.HpaI. All five transformed lines were found to contain DNA sequences homologous to 75--80% of Ad5 fragment HsuI G only. Clones II and V contained approximately 48 copies per quantity of diploid cell DNA, clone VI about 35 copies, clone IV 22 copies and clone III 5--10 copies. These results indicate that a viral DNA segment as small as 5.5% of the Ad5 genome, contains sufficient information for the maintenance of transformation.
对五个由腺病毒5(Ad5)DNA的一个小限制性内切酶片段(HsuI G片段,代表腺病毒基因组的左末端7%)转化的大鼠肾细胞克隆,分析了细胞DNA中存在的病毒DNA序列。在这些分析中,在存在从每个转化细胞系或未转化细胞中提取的大量DNA的情况下,测量了32P标记的Ad5 DNA特异性片段的复性动力学。这些片段是通过用内切酶R.HsuI消化32P标记的腺病毒5 DNA,或用内切酶R.HpaI消化32P标记的腺病毒5 DNA的HsuI G片段产生的。发现所有五个转化系仅含有与Ad5 HsuI G片段的75 - 80%同源的DNA序列。克隆II和V每二倍体细胞DNA量含有约48个拷贝,克隆VI约35个拷贝,克隆IV 22个拷贝,克隆III 5 - 10个拷贝。这些结果表明,一个小至Ad5基因组5.5%的病毒DNA片段含有维持转化所需的足够信息。