Collins C J, Parsons J T
Proc Natl Acad Sci U S A. 1977 Oct;74(10):4301-5. doi: 10.1073/pnas.74.10.4301.
DNA from six avian sarcoma virus (ASV)-transformed mammalian cell lines was digested with the restriction endonucleases EcoRI, Xho I, or Sal I, fractionated by agarose gel electrophoresis, transferred to nitrocellulose filter strips, and hybridized with specific ASV [32P]cDNA probes. DNA from all of the ASV-transformed cell lines yielded three common virus-specific DNA fragments (2.4, 1.8, and 1.3 X 10(6) daltons) upon cleavage with EcoRI. Xho I appeared to cleave at least once within the integrated provirus and yielded a common fragment of 3.3 X 10(6) daltons as well as a second virus-specific DNA fragment whose size varied from 4.0 to 5.0 X 10(6) daltons in the different transformed cell lines. Sal I did not cleave within the provirus and yielded a single major virus-specific fragment of about 11 X 10(6) daltons in all transformed lines examined. Using specific cDNA probes, we show that the 1.8 X 10(6)-dalton EcoRI fragment contains sequences homologous to the 3' end of the viral RNA as well as to the src region of the viral genome. These studies clearly demonstrate that the same region on the ASV genome is utilized for provirus integration in different ASV-transformed cell lines.
用限制性内切酶EcoRI、Xho I或Sal I消化来自六种禽肉瘤病毒（ASV）转化的哺乳动物细胞系的DNA，通过琼脂糖凝胶电泳进行分离，转移至硝酸纤维素滤纸条上，并用特异性ASV [32P] cDNA探针进行杂交。用EcoRI切割后，来自所有ASV转化细胞系的DNA产生了三个常见的病毒特异性DNA片段（2.4、1.8和1.3×10⁶道尔顿）。Xho I似乎在整合的前病毒内至少切割一次，并产生了一个3.3×10⁶道尔顿的常见片段以及第二个病毒特异性DNA片段，其大小在不同的转化细胞系中从4.0到5.0×10⁶道尔顿不等。Sal I在前病毒内不切割，在所检测的所有转化细胞系中产生了一个约11×10⁶道尔顿的单一主要病毒特异性片段。使用特异性cDNA探针，我们表明1.8×10⁶道尔顿的EcoRI片段包含与病毒RNA 3'端以及病毒基因组src区域同源的序列。这些研究清楚地表明，ASV基因组上的相同区域用于不同ASV转化细胞系中的前病毒整合。
