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阿留申病病毒基因组及其细胞内形式的特征分析。

Characterization of the Aleutian disease virus genome and its intracellular forms.

作者信息

Bloom M E, Mayer L W, Garon C F

出版信息

J Virol. 1983 Mar;45(3):977-84. doi: 10.1128/JVI.45.3.977-984.1983.

Abstract

Aleutian disease virus (ADV) of mink is a nondefective parvovirus with a single-stranded DNA genome. We characterized the viral DNA forms found in infected cells prepared by a modified Hirt extraction procedure. Double-stranded DNA molecules corresponding in size to 4.8-kilobase-pair duplex monomers and 9.6-kilobase-pair duplex dimers were identified in agarose gels by blot hybridization to 32P-labeled ADV DNA. A rapidly reannealing ADV duplex monomer was isolated on a preparative scale and physically mapped with a series of restriction endonucleases. The map derived was similar to one derived from double-stranded ADV DNA produced by self-primed synthesis on virion DNA, but differed from restriction endonuclease maps reported for other parvovirus DNAs. The purified duplex monomer could be labeled with [32P]dCTP by nick translation and used as a probe in blot hybridization to detect ADV sequences in DNA from small numbers of infected cells. Additional studies indicated that double-stranded ADV DNA could first be detected at 24 h after infection.

摘要

水貂阿留申病病毒(ADV)是一种具有单链DNA基因组的非缺陷型细小病毒。我们对通过改良的赫特提取程序制备的感染细胞中发现的病毒DNA形式进行了表征。通过与32P标记的ADV DNA进行印迹杂交,在琼脂糖凝胶中鉴定出大小对应于4.8千碱基对双链单体和9.6千碱基对双链二聚体的双链DNA分子。在制备规模上分离出一种快速复性的ADV双链单体,并用一系列限制性内切酶进行物理图谱绘制。得到的图谱与通过在病毒体DNA上进行自我引物合成产生的双链ADV DNA图谱相似,但与报道的其他细小病毒DNA的限制性内切酶图谱不同。纯化的双链单体可以通过缺口平移用[32P]dCTP进行标记,并用作印迹杂交中的探针,以检测来自少量感染细胞的DNA中的ADV序列。进一步的研究表明双链ADV DNA在感染后24小时首次被检测到。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a7ba/256504/4c996f9ae15a/jvirol00150-0089-a.jpg

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