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溶液中和凝胶中闭环DNA的超螺旋圈数:构象差异的证据。

Superhelical turns of closed circular DNA in solution and in a gel: evidence for a conformational difference.

作者信息

Dougherty G

出版信息

Aust J Exp Biol Med Sci. 1982 Dec;60(6):675-86. doi: 10.1038/icb.1982.69.

Abstract

We have compared the number of superhelical turns, tau, in circular covalently closed plasmid pBR beta G DNA obtained by four different methods, each based on one particular distinguishing principle. Three of the methods allow an unequivocal determination of tau under gel electrophoresis conditions, whilst the fourth enables us to determine its value in solution. We were able to detect a significant difference between the two environments, corresponding to an unwinding of the DNA duplex angle by 0.3 degrees when a sample is transferred from solution to gel. The possible existence of such an effect has been generally overlooked by previous investigators. Our result suggests that the previously reported value for the number of base-pairs per helical turn should be adjusted downwards by about 0.10, so that it applies to conditions in solution.

摘要

我们比较了通过四种不同方法获得的环状共价闭合质粒pBRβG DNA中的超螺旋圈数τ,每种方法都基于一个特定的区分原理。其中三种方法可以在凝胶电泳条件下明确测定τ,而第四种方法使我们能够在溶液中测定其值。我们能够检测到两种环境之间的显著差异,当样品从溶液转移到凝胶时,这相当于DNA双链角度解开0.3度。以前的研究人员普遍忽略了这种效应可能的存在。我们的结果表明,先前报道的每螺旋圈碱基对数的值应向下调整约0.10,以便适用于溶液中的条件。

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