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[丁酸钠和鲁米诺对培养动物细胞染色体外DNA重组过程中相互交换和基因转换的影响]

[The effect of sodium butyrate and luminol on reciprocal exchanges and gene conversion during extrachromosomal DNA recombination in cultured animal cells].

作者信息

Glebov O K, Abramian D S, Romanov S R, Smagina L V

出版信息

Tsitologiia. 1994;36(5):441-52.

PMID:7809980
Abstract

For determination of the extrachromosomal homologous DNA recombination efficiency, somatic cells of various lines have been transformed with plasmid DNAs which contain copies of neo-gene with non-overlapping deletions. Reconstruction of the neo-gene functional activity, which imparts a geneticin-resistant phenotype to cells, indicates that recombination has occurred. If dP1 and dR copies of the neo-gene are used, a single (reciprocal) exchange is necessary for reconstruction of the neo-gene by homologous DNA recombination, but a double exchange (gene conversion) is needed in the case of dP1 and dS copies. It is shown that in human cells of line HeLa and in mouse cells of line LMtk-, in contrast to the Chinese hamster cells of line A238, the frequency of double exchanges is comparable to that of the single DNA exchanges which is an evidence of participation in DNA recombination of gene conversion in addition to a single exchange mechanism. The treatment of cells with sodium butyrate and luminol exerts different influences on the rate of the single DNA exchanges and on that of gene conversion (double exchanges) in cells of lines LMtk- and HeLa, respectively. Essential distinctions in correlation of the single DNA exchange frequency and the gene conversion frequency in cells of the studied lines, and the possibility to distinguish between these mechanisms of recombination, under the treatment by sodium butyrate and luminol, may suggest the existence of two mechanisms of homologous DNA recombination in cultured animal cells, which function independently of one another, to a considerable extent.

摘要

为了测定染色体外同源DNA重组效率,用含有带有非重叠缺失的新霉素基因拷贝的质粒DNA转化了各种细胞系的体细胞。新霉素基因功能活性的重建赋予细胞遗传霉素抗性表型,这表明发生了重组。如果使用新霉素基因的dP1和dR拷贝,通过同源DNA重组重建新霉素基因需要一次(相互)交换,但对于dP1和dS拷贝,则需要两次交换(基因转换)。结果表明,与A238细胞系的中国仓鼠细胞相比,HeLa细胞系的人细胞和LMtk-细胞系的小鼠细胞中,两次交换的频率与单DNA交换的频率相当,这证明除了单交换机制外,基因转换也参与了DNA重组。用丁酸钠和鲁米诺处理细胞,分别对LMtk-和HeLa细胞系的细胞中单DNA交换速率和基因转换(两次交换)速率产生不同影响。在所研究细胞系的细胞中,单DNA交换频率与基因转换频率的相关性存在本质区别,并且在丁酸钠和鲁米诺处理下能够区分这些重组机制,这可能表明在培养的动物细胞中存在两种同源DNA重组机制,它们在很大程度上相互独立发挥作用。

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