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论DNA链断裂作为引发姐妹染色单体交换(SCE)的首个事件的重要性:使用限制性内切酶BglI的实验

On the importance of DNA strand breaks as the first event to initiate sister chromatid exchange (SCE): experiments with restriction endonuclease BglI.

作者信息

Ortiz T, Piñero J, Cortés F

机构信息

Department of Cell Biology, Faculty of Biology, Seville, Spain.

出版信息

Chromosome Res. 1996 Nov;4(7):540-4. doi: 10.1007/BF02261782.

Abstract

The exact molecular mechanism of sister chromatid exchange (SCE) is still unknown, despite the many reports dealing with this cytogenetic end point published in the last 40 years. One point to be investigated is the nature of the original lesion(s) in DNA leading to the production of SCE. Whereas, for chromosomal aberrations, the importance of DNA double-strand breaks has been well established, there is still controversy about the relative importance of strand breaks and base modifications for triggering the process of SCE formation. In the present paper, we have taken advantage of the ability of the restriction endonuclease BglI to induce SCE and have exploited the fact that preincubation with 2,3-butanedione results in the loss of BgiI ability to cut DNA, while it is still able to recognize its sequence in DNA and bind to it, to see whether this alone is enough to initiate SCE formation, or if a physical DNA double-strand break is required. Our results seem to support the necessity of DNA breaks for SCE production.

摘要

尽管在过去40年里发表了许多关于这个细胞遗传学终点的报告,但姐妹染色单体交换(SCE)的确切分子机制仍然未知。一个需要研究的问题是导致SCE产生的DNA原始损伤的性质。对于染色体畸变,DNA双链断裂的重要性已经得到充分证实,然而,关于链断裂和碱基修饰在触发SCE形成过程中的相对重要性仍存在争议。在本文中,我们利用限制性内切酶BglI诱导SCE的能力,并利用了这样一个事实:与2,3 - 丁二酮预孵育会导致BglI切割DNA的能力丧失,而它仍然能够识别DNA中的序列并与之结合,以此来观察这是否足以启动SCE的形成,或者是否需要物理性的DNA双链断裂。我们的结果似乎支持了DNA断裂对于SCE产生的必要性。

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