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疣猴C型病毒:未整合病毒DNA的分子克隆及疣猴内源性病毒基因组的特征分析

Colobus type C virus: molecular cloning of unintegrated viral DNA and characterization of the endogenous viral genomes of Colobus.

作者信息

Birkenmeier E H, Bonner T I, Reynolds K, Searfoss G H, Todaro G J

出版信息

J Virol. 1982 Mar;41(3):842-54. doi: 10.1128/JVI.41.3.842-854.1982.

Abstract

The unintegrated viral DNA intermediates of colobus type C virus (CPC-1) were isolated from infected human cells that were permissive for viral growth. There were two major species of DNA, linear molecules with two copies of the long terminal repeat and relaxed circles containing only a single long terminal repeat. In addition, there was a minor species (approximately 10%) composed of relaxed circles with two copies of the long terminal repeat. A restriction endonuclease map of the unintegrated DNA was constructed. The three EcoRI fragments of circular CPC-1 DNA were cloned in the EcoRI site of lambda gtWES . lambda B and then subcloned in the EcoRI site of pBR322. Using these subgenomic fragments as probes, we have characterized the endogenous viral sequences found in colobus cellular DNA. They are not organized in tandem arrays, as is the case in some other gene families. The majority of sequences detected in cellular DNA have the same map as the CPC-1 unintegrated DNA at 17 of 18 restriction endonuclease sites. There are, however, other sequences that are present in multiple copies and do not correspond to the CPC-1 map. They do not contain CPC-1 sequences either in an altered form or fused to common nonviral sequences. Instead, they appear to be derived from a distinct family of sequences that is substantially diverged from the CPC-1 family. This second family of sequences, CPC-2, is also different from the sequences related to baboon endogenous type C virus that forms a third family of virus-related sequences in the colobus genome.

摘要

从允许病毒生长的受感染人类细胞中分离出疣猴C型病毒(CPC - 1)的未整合病毒DNA中间体。有两种主要的DNA种类,一种是带有两个长末端重复序列拷贝的线性分子,另一种是仅含有一个长末端重复序列的松弛环状分子。此外,还有一小部分(约10%)由带有两个长末端重复序列拷贝的松弛环状分子组成。构建了未整合DNA的限制性内切酶图谱。环状CPC - 1 DNA的三个EcoRI片段被克隆到λgtWES.λB的EcoRI位点,然后亚克隆到pBR322的EcoRI位点。使用这些亚基因组片段作为探针,我们对疣猴细胞DNA中发现的内源性病毒序列进行了表征。它们不像其他一些基因家族那样以串联阵列的形式组织。在细胞DNA中检测到的大多数序列在18个限制性内切酶位点中的17个处与CPC - 1未整合DNA具有相同的图谱。然而,还有其他以多拷贝形式存在且与CPC - 1图谱不对应的序列。它们既不包含改变形式的CPC - 1序列,也不与常见的非病毒序列融合。相反,它们似乎源自一个与CPC - 1家族有很大差异的独特序列家族。这个第二个序列家族,即CPC - 2,也不同于与狒狒内源性C型病毒相关的序列,后者在疣猴基因组中形成了第三个病毒相关序列家族。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/260b/256821/0ecbb5e63d60/jvirol00162-0110-a.jpg

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