人类淋巴母细胞系中细胞DNA与爱泼斯坦-巴尔病毒DNA之间的线性关联。
Linear association between cellular DNA and Epstein-Barr virus DNA in a human lymphoblastoid cell line.
作者信息
Adams A, Lindahl T, Klein G
出版信息
Proc Natl Acad Sci U S A. 1973 Oct;70(10):2888-92. doi: 10.1073/pnas.70.10.2888.
High-molecular-weight DNA from cell line Raji (derived from Burkitt's lymphoma), which contains 50-60 copies of Epstein-Barr virus DNA per cell, was fractionated in neutral solution by several cycles of CsCl gradient centrifugation in fixed-angle rotors. Under the fractionation conditions used, intact Epstein-Barr virus DNA from virus particles can be separated from the less-dense cellular DNA. In contrast, a large proportion of the intrinsic Epstein-Barr virus DNA component of Raji cells remains associated with cellular DNA, as determined by nucleic acid hybridization. This interaction, which is resistant to Pronase and phenol treatment, is not the result of aggregation. When the molecular weight of Raji DNA is reduced by hydrodynamic shear, the amount of virus DNA associated with cell DNA decreases. However, some virus DNA still remains bound to fragments of cellular DNA after shearing. The association is completely destroyed in alkaline solution. Molecular weight analysis of Raji DNA after denaturation showed that the alkali-induced release of Epstein-Barr virus DNA was specific and not the result of random single-strand breaks. These data indicate that Epstein-Barr virus DNA is linearly integrated into Raji cell DNA by alkali-labile bonds.
来自Raji细胞系（源自伯基特淋巴瘤）的高分子量DNA，每个细胞含有50 - 60份爱泼斯坦-巴尔病毒DNA，通过在固定角度转子中进行几个循环的CsCl梯度离心在中性溶液中分级分离。在所使用的分级分离条件下，来自病毒颗粒的完整爱泼斯坦-巴尔病毒DNA可以与密度较低的细胞DNA分离。相比之下，通过核酸杂交测定，Raji细胞的大部分内源性爱泼斯坦-巴尔病毒DNA成分仍与细胞DNA相关联。这种相互作用对蛋白酶和苯酚处理具有抗性，并非聚集的结果。当Raji DNA的分子量通过流体动力学剪切降低时，与细胞DNA相关联的病毒DNA量减少。然而，剪切后仍有一些病毒DNA与细胞DNA片段结合。这种关联在碱性溶液中完全被破坏。变性后Raji DNA的分子量分析表明，碱诱导的爱泼斯坦-巴尔病毒DNA释放是特异性的，并非随机单链断裂的结果。这些数据表明，爱泼斯坦-巴尔病毒DNA通过碱不稳定键线性整合到Raji细胞DNA中。
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