EPSTEIN-BARR VIRUS GENOMES WITH PROPERTIES OF CIRCULAR DNA-MOLECULES IN CARRIER CELLS
EPSTEIN-BARR VIRUS GENOMES WITH PROPERTIES OF CIRCULAR DNA-MOLECULES IN CARRIER CELLS
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DOI：
10.1073/pnas.72.4.1477
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发表时间：
1975-01-01
影响因子：
11.1
通讯作者：
LINDAHL, T
中科院分区：
文献类型：
--
作者：
ADAMS, A;LINDAHL, T
从人淋巴样细胞系 Raji 中分离出高密度的高分子量 DNA。该细胞系每个细胞含有 50 至 60 个与 Epstein-Barr 病毒 DNA 等价的病毒基因组，并且此类病毒序列中的高密度 DNA 部分富集了 10 倍。中性甘油梯度的沉降分析，随后与病毒互补RNA的杂交实验表明，该材料中的大多数细胞内病毒DNA序列不与细胞DNA共沉降，而是作为两种不同的物种回收，沉降系数分别为100 S和65 S。这两种形式的沉降速度是病毒颗粒中线性Epstein-Barr病毒DNA的1.70-1.75和1.10-1.12倍，因此具有病毒基因组的共价闭合环状形式和带切口（包含单链断裂）环状形式的流体动力学特性。在 CsCl/二碘化丙啶梯度离心中，100S 形式也表现为共价闭合环状 EBV DNA 分子。看来潜伏的 Epstein-Barr 病毒 DNA 具有哺乳动物附加体的特性，并且非整合和整合的病毒 DNA 序列都可以从载体细胞中分离出来。
A high-density fraction of high-molecular-weight DNA was isolated from the human lymphoid cell line Raji. This cell line contains 50 to 60 virus genome equivalents of Epstein-Barr virus DNA per cell, and the high-density DNA fraction was 10-fold enriched in such viral sequences. Sedimentation analysis on neutral glycerol gradients, followed by hybridization experiments with viral complementary RNA, showed that most of the intracellular viral DNA sequences in this material did not cosediment with the cellular DNA, but were recovered as two distinct species with sedimentation coefficients of 100 S and 65 S. These two forms sediment 1.70-1.75 and 1.10-1.12 times as fast as the linear Epstein-Barr virus DNA from virus particles, and thus have the hydrodynamic properties of a covalently closed circular form and a nicked (containing single-strand breaks) circular form of the virus genome. The 100S form also behaved as a covalently closed circular EBV DNA molecule on gradient centrifugation in CsC1/propidium diiodide, It would appear that latent Epstein-Barr virus DNA has the properties of a mammalian episome, and that both nonintegrated and integrated viral DNA sequences can be isolated from carrier cells.