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序列特异性超声 DNA 切割。

Sequence-specific ultrasonic cleavage of DNA.

机构信息

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia.

出版信息

Biophys J. 2011 Jan 5;100(1):117-25. doi: 10.1016/j.bpj.2010.10.052.

Abstract

We investigated the phenomenon of ultrasonic cleavage of DNA by analyzing a large set of cleavage patterns of DNA restriction fragments using polyacrylamide gel electrophoresis. The cleavage intensity of individual phosphodiester bonds was found to depend on the nucleotide sequence and the position of the bond with respect to the ends of the fragment. The relative intensities of cleavage of the central phosphodiester bond in 16 dinucleotides and 256 tetranucleotides were determined by multivariate statistical analysis. We observed a remarkable enhancement of the mean values of the relative intensities of cleavage (cleavage rates) in phosphodiester bonds following deoxycytidine, which diminished in the row of dinucleotides: d(CpG) > d(CpA) > d(CpT) >> d(CpC). The cleavage rates for all pairs of complementary dinucleotides were significantly different from each other. The effect of flanking nucleotides in tetranucleotides on cleavage rates of all 16 types of central dinucleotides was also statistically significant. The sequence-dependent ultrasonic cleavage rates of dinucleotides are consistent with reported data on the intensity of the conformational motion of their 5'-deoxyribose. As a measure of local conformational dynamics, cleavage rates may be useful for characterizing functional regions of the genome.

摘要

我们通过分析大量使用聚丙烯酰胺凝胶电泳的 DNA 限制片段的切割模式来研究超声切割 DNA 的现象。发现单个磷酸二酯键的切割强度取决于核苷酸序列以及键相对于片段末端的位置。通过多元统计分析确定了 16 个二核苷酸和 256 个四核苷酸中中央磷酸二酯键的相对切割强度。我们观察到在脱氧胞嘧啶之后,磷酸二酯键的相对切割强度(切割率)的平均值显着增强,在二核苷酸行中减小:d(CpG) > d(CpA) > d(CpT) >> d(CpC)。所有互补二核苷酸对的切割率彼此显着不同。四核苷酸中外围核苷酸对所有 16 种中心二核苷酸切割率的影响在统计学上也具有显着性。二核苷酸的序列依赖性超声切割率与它们 5'-脱氧核糖构象运动强度的报道数据一致。作为局部构象动力学的度量,切割率可能有助于表征基因组的功能区域。

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