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回文DNA研究。

A study of foldback DNA.

作者信息

Perlman S, Phillips C, Bishop J O

出版信息

Cell. 1976 May;8(1):33-42. doi: 10.1016/0092-8674(76)90182-3.

Abstract

Nuclear DNA from eucaryotes contains a significant fraction which forms duplexes very rapidly and also independently of the DNA concentration. This fraction can be isolated by adsorption to hydroxylapatite and has been called foldback DNA (Britten and Smith, 1970). Here we extend previous studies to show that the foldback fraction is due to the existence of a finite number of foldback foci in each genome equivalent of DNA, approximately 10(5) in the case of Xenopus laevis. More significantly, we have isolated the foldback fraction in quantity from DNA of such a size (in one case broken randomly and in another digested with a restriction endonuclease) that only about 10% of the total DNA has foldback properties. If the foldback foci were located in precisely the same positions in all sets of the Xenopus laevis genome, the prediction would be that these foldback fractions would contain sequences representing 20% (random shear) and 10% (restriction endonuclease) of the total genome. In contrast, our results show that in both cases the foldback fraction contains the entire Xenopus laevis DNA sequence. One possible explanation of these observations is that as in procaryotes, eucaryotic DNA is randomly cross-linked. We show that cross-linkage of Xenopus laevis DNA is not sufficient to explain our observations. In consequence, we have adopted the hypothesis that the formation of foldback DNA is mainly an intrastrand phenomenon, but nevertheless occurs at different sites in different sets of the Xenopus laevis genome.

摘要

真核生物的核DNA中有很大一部分能非常迅速地形成双链,而且这一过程与DNA浓度无关。这部分DNA可以通过吸附到羟基磷灰石上分离出来,被称为回折DNA(布里顿和史密斯,1970年)。在此,我们扩展了先前的研究,以表明回折部分是由于每个基因组当量的DNA中存在有限数量的回折位点,在非洲爪蟾的情况下大约有10⁵个。更重要的是,我们从如此大小的DNA(一种情况是随机断裂,另一种情况是用限制性内切酶消化)中大量分离出了回折部分,以至于只有大约10%的总DNA具有回折特性。如果回折位点在非洲爪蟾基因组的所有组中都精确地位于相同位置,那么预测结果将是这些回折部分将包含代表整个基因组20%(随机剪切)和10%(限制性内切酶)的序列。相比之下,我们的结果表明,在这两种情况下,回折部分都包含了整个非洲爪蟾的DNA序列。对这些观察结果的一种可能解释是,与原核生物一样,真核生物的DNA是随机交联的。我们表明,非洲爪蟾DNA的交联不足以解释我们的观察结果。因此,我们采用了这样一种假说,即回折DNA的形成主要是一种链内现象,但仍然发生在非洲爪蟾基因组不同组的不同位点上。

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