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外源DNA片段对人细胞提取物介导的DNA修复合成的影响。

Effect of exogenous DNA fragments on human cell extract-mediated DNA repair synthesis.

作者信息

Biggerstaff M, Robins P, Coverley D, Wood R D

机构信息

Imperial Cancer Research Fund, Clare Hall Laboratories, South Mimms, Herts, Great Britain.

出版信息

Mutat Res. 1991 May;254(3):217-24. doi: 10.1016/0921-8777(91)90059-x.

DOI:10.1016/0921-8777(91)90059-x
PMID:2052011
Abstract

Extracts from HeLa cells were used to study the susceptibility of repair synthesis in UV-irradiated plasmid DNA to inhibition by exogenously added nucleic acid. Purified DNA restriction fragments have little inhibitory effect on repair synthesis. However, activated calf thymus DNA fragments, genomic DNA fragments in cell extracts, and sonicated plasmid DNA all inhibited repair synthesis. Degraded DNA fragments arising from E. coli during bacterial plasmid purification were found to be particularly inhibitory. tRNA is not a potent inhibitor of in vitro repair synthesis. In order to observe efficient DNA repair synthesis mediated by human cell extracts, it is essential to prepare highly purified closed circular plasmid DNA, and we describe a reliable method for doing so.

摘要

利用从海拉细胞中提取的物质来研究紫外线照射过的质粒DNA中修复合成对外源添加核酸抑制作用的敏感性。纯化的DNA限制片段对修复合成几乎没有抑制作用。然而，活化的小牛胸腺DNA片段、细胞提取物中的基因组DNA片段以及超声处理的质粒DNA均能抑制修复合成。发现在细菌质粒纯化过程中由大肠杆菌产生的降解DNA片段具有特别强的抑制作用。tRNA不是体外修复合成的有效抑制剂。为了观察由人类细胞提取物介导的高效DNA修复合成，制备高度纯化的闭环质粒DNA至关重要，我们描述了一种可靠的制备方法。

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Reduced RNA polymerase II transcription in extracts of cockayne syndrome and xeroderma pigmentosum/Cockayne syndrome cells.
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