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通过用乙二醛完全封闭鸟嘌呤残基来防止小鼠DNA中G:C配对。胞嘧啶、腺嘌呤和胸腺嘧啶与添加的未修饰多核苷酸形成氢键的可能性。

Prevention of G:C pairing in mouse DNA by complete blocking of guanine residues with glyoxal. Availability of cytosine, adenine and thymine for hydrogen bonding with added unmodified polynucleotides.

作者信息

Birnboim H C, Mitchel R E

出版信息

Biochim Biophys Acta. 1978 Feb 16;517(2):296-307. doi: 10.1016/0005-2787(78)90196-x.

Abstract

We have developed conditions for the reaction of single-stranded DNA with glyoxal which permit blocking of essentially all guanine residues. This procedure effectively prevents base pairing involving these guanine residues, yet permits cytosine, thymine and adenine residues in the DNA to pair with added polynucleotides. Modification of DNA with glyoxal greatly reduces intrastrand helical regions, resulting in a very low binding to hydroxyapatite, as compared to unmodified DNA. Annealing of modified DNA with some synthetic polynucleotides of restricted composition (but not others) leads to a significant increase in binding, presumably because mouse DNA has sequences not containing guanine which are capable of hydrogen bonding to the added polynucleotides. This relatively simple procedure may allow isolation and further study of these guanine-free sequences in DNA.

摘要

我们已经开发出了单链DNA与乙二醛反应的条件,该条件能使基本上所有的鸟嘌呤残基被封闭。此程序有效地阻止了涉及这些鸟嘌呤残基的碱基配对,但允许DNA中的胞嘧啶、胸腺嘧啶和腺嘌呤残基与添加的多核苷酸配对。与未修饰的DNA相比,用乙二醛修饰DNA会大大减少链内螺旋区域,导致与羟基磷灰石的结合非常低。用一些组成受限的合成多核苷酸(但不是其他多核苷酸)对修饰后的DNA进行退火会导致结合显著增加,推测是因为小鼠DNA具有不包含鸟嘌呤的序列,这些序列能够与添加的多核苷酸形成氢键。这个相对简单的程序可能允许分离并进一步研究DNA中这些不含鸟嘌呤的序列。

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