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狒狒内源性病毒基因组:来自狒狒细胞系DNA的无缺陷病毒基因组的分子克隆与结构表征

Baboon endogenous virus genome: molecular cloning and structural characterization of nondefective viral genomes from DNA of a baboon cell strain.

作者信息

Cohen M, Rein A, Stephens R M, O'Connell C, Gilden R V, Shure M, Nicolson M O, McAllister R M, Davidson N

出版信息

Proc Natl Acad Sci U S A. 1981 Aug;78(8):5207-11. doi: 10.1073/pnas.78.8.5207.

Abstract

Several heterogeneities in the baboon endogenous virus (BaEV) genomes that are present in the DNA of normal baboon tissues and the baboon cell strain BEF-3 have been described previously. To study these genomes, we cloned BaEV proviruses from BEF-3 cellular DNA into the lambda vector Charon 4A. Of the four full-length clones isolated, one was nondefective as determined by transfection. The sequence of a portion of this clone was found to code for amino acids 61-91 in the p30 region of the gag gene. This identification allowed us to align the restriction map with the BaEV genetic map. One heterogeneity, a BamHI site 2.4 kilobases (kb) from the proviral 5' end, was located close to the gag-pol junction; another, a BamHI site 1.4 kb from the 5' end of the genome, corresponded to the gag p30 coding sequence for amino acids 32-34; and a third, a Xho I site, was near the 3' end of the pol gene. To select the nondefective BaEV genomes from BEF-3 cells, we infected permissive cells with virus produced by BEF-3 cells and also transfected BEF-3 cellular DNA into permissive cells. The BaEV genomes in the permissive recipient cultures were then analyzed by restriction enzyme analysis. These nondefective genomes were found to be heterogeneous with respect to the gag-pol BamHI site and the Xho I site, but all were found to contain the BamHI site 1.4 kb from the 5' end of the genome.

摘要

先前已描述过正常狒狒组织的DNA和狒狒细胞系BEF-3中存在的狒狒内源性病毒(BaEV)基因组中的几种异质性。为了研究这些基因组,我们将来自BEF-3细胞DNA的BaEV前病毒克隆到λ载体Charon 4A中。在分离出的四个全长克隆中,有一个经转染测定为无缺陷。发现该克隆一部分的序列编码gag基因p30区域中第61至91位氨基酸。这一鉴定使我们能够将限制酶图谱与BaEV遗传图谱进行比对。一种异质性是,位于前病毒5'端2.4千碱基(kb)处的一个BamHI位点,靠近gag-pol连接点;另一种是,位于基因组5'端1.4 kb处的一个BamHI位点,对应于gag p30编码第32至34位氨基酸的序列;第三种是,一个Xho I位点,位于pol基因的3'端附近。为了从BEF-3细胞中选择无缺陷的BaEV基因组,我们用BEF-3细胞产生的病毒感染允许性细胞,并将BEF-3细胞DNA转染到允许性细胞中。然后通过限制酶分析对允许性受体培养物中的BaEV基因组进行分析。发现这些无缺陷的基因组在gag-pol BamHI位点和Xho I位点方面存在异质性,但均发现含有位于基因组5'端1.4 kb处的BamHI位点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/05a5/320375/7a89afe868d2/pnas00659-0585-a.jpg

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