Markham P D, Baluda M A
J Virol. 1973 Oct;12(4):721-32. doi: 10.1128/JVI.12.4.721-732.1973.
The covalent linkage of oncornavirus-specific DNA to chicken DNA was investigated in normal chicken embryo fibroblasts (CEF) and in virus-producing leukemic cells transformed by avian myeloblastosis virus (AMV). The virus-specific sequences present in cellular DNA fractionated by different methods were detected by DNA-RNA hybridization by using 70S AMV RNA as a probe. In CEF and in leukemic cells, the viral DNA appeared to be present only in the nucleus. After cesium chloride-ethidium bromide density equilibrium sedimentation, the viral DNA was present as linear, double-stranded molecules not separable from linear chicken DNA. After extraction by the Hirt procedure, the viral DNA precipitated with the high-molecular-weight DNA. After alkaline sucrose velocity sedimentation, the viral DNA cosedimented with the high-molecular-weight cellular DNA. The results indicate that in both types of cells studied, the oncornavirus-specific DNA sequences were linked by alkali stable bonds to nuclear cellular DNA of high molecular weight and did not appear to be present in free form of any size.
在正常鸡胚成纤维细胞(CEF)以及由禽成髓细胞瘤病毒(AMV)转化产生病毒的白血病细胞中,研究了肿瘤病毒特异性DNA与鸡DNA的共价连接。通过使用70S AMV RNA作为探针进行DNA - RNA杂交,检测了通过不同方法分级分离的细胞DNA中存在的病毒特异性序列。在CEF和白血病细胞中,病毒DNA似乎仅存在于细胞核中。经过氯化铯 - 溴化乙锭密度平衡沉降后,病毒DNA以线性双链分子形式存在,无法与线性鸡DNA分离。通过Hirt方法提取后,病毒DNA与高分子量DNA一起沉淀。经过碱性蔗糖速度沉降后,病毒DNA与高分子量细胞DNA共同沉降。结果表明,在所研究的两种细胞类型中,肿瘤病毒特异性DNA序列通过碱稳定键与高分子量的细胞核细胞DNA相连,并且似乎不以任何大小的游离形式存在。