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[体细胞的遗传转化。III. 转化体克隆细胞染色体外DNA中质粒核苷酸序列状态的分析及质粒DNA染色体外分子的拯救]

[Genetic transformation of somatic cells. III. An analysis of the status of the plasmid nucleotide sequences in the extrachromosomal DNA of transformant clone cells and the rescue of extrachromosomal molecules of the plasmid DNA].

作者信息

Tomilin N V, Svetlova M P, Podgornaia O I, Glebov O K

出版信息

Tsitologiia. 1985 May;27(5):565-71.

PMID:2990075
Abstract

Extrachromosomal DNAs from TK+ transformant clones of A238 Chinese hamster cells isolated after the treatment with plasmid pST826 containing thymidine kinase gene (TK-gene) of Herpes simplex virus (HSV1) and 1.8 kb insert of human satellite III DNA (HSIII) were studied by hybridization technique. In two TK+-clones (2T301 and 2T16) large quantities of rearranged plasmid DNA molecules were found. Electron microscopy show in clone 2T301 the presence of circular DNAs with average length being 4.64 +/- 0.27 kb. These molecules were rescued by retransformation into E. coli and analysed by restriction mapping and hybridization. All of them contain deletions spanning the entire TK gene of HSV1 and pBR325 sequences situated just downstream from the ORI of replication. The origin of extra-replicating circular DNA in 2T301 clone is discussed.

摘要

用含有单纯疱疹病毒1型(HSV1)胸苷激酶基因(TK基因)和1.8 kb人卫星III DNA(HSIII)插入片段的质粒pST826处理后分离得到的A238中国仓鼠细胞的TK +转化体克隆中的染色体外DNA,通过杂交技术进行了研究。在两个TK +克隆(2T301和2T16)中发现了大量重排的质粒DNA分子。电子显微镜显示在克隆2T301中存在平均长度为4.64 +/- 0.27 kb的环状DNA。这些分子通过重新转化到大肠杆菌中进行拯救,并通过限制性图谱分析和杂交进行分析。它们都包含跨越HSV1整个TK基因的缺失以及位于复制起点下游的pBR325序列。讨论了2T301克隆中额外复制环状DNA的起源。

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