Schick J, Doerfler W
J Gen Virol. 1978 May;39(2):365-70. doi: 10.1099/0022-1317-39-2-365.
In human KB cells permissively infected with adenovirus type 2, the high mol. wt. forms of virus DNA have been characterized. These size classes of virus DNA sediment at greater than 100S and 40 to 90S in alkaline sucrose density gradients. Considerable evidence from a series of earlier communications supports the notion that the high mol. wt. forms of virus DNA represent virus DNA sequences covalently linked to cellular DNA. 3H-labelled high mol. wt. adenovirus type 2 DNA from productively infected cells can be shown to hybridize to virus DNA fixed to filters. In the present paper we demonstrate that on alkali elution of the DNA from the filters used in the first step of the hybridization experiment, the labelled DNA re-hybridizes to cellular DNA in the second step of a sequential hybridization experiment. The order of performing the two successive hybridization experiments can be reversed and very similar results are obtained. These data provide conclusive evidence for the covalent linkage of virus and cellular DNA sequences in KB cells productively infected with adenovirus type 2.
在被2型腺病毒允许性感染的人KB细胞中，已对高分子量的病毒DNA形式进行了表征。这些大小类别的病毒DNA在碱性蔗糖密度梯度中以大于100S和40至90S的速度沉降。一系列早期通讯中的大量证据支持这样一种观点，即高分子量的病毒DNA形式代表与细胞DNA共价连接的病毒DNA序列。来自生产性感染细胞的3H标记的高分子量2型腺病毒DNA可显示与固定在滤膜上的病毒DNA杂交。在本文中，我们证明，在杂交实验第一步所用滤膜上的DNA进行碱洗脱后，标记的DNA在连续杂交实验的第二步中与细胞DNA重新杂交。进行这两个连续杂交实验的顺序可以颠倒，并且会获得非常相似的结果。这些数据为在被2型腺病毒生产性感染的KB细胞中病毒和细胞DNA序列的共价连接提供了确凿证据。
