Peptide nucleic acid-assisted topological labeling of duplex DNA
Peptide nucleic acid-assisted topological labeling of duplex DNA
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DOI：
10.1006/meth.2000.1113
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发表时间：
2001-02-01
期刊：
影响因子：
4.8
通讯作者：
Frank-Kameneteskii, MD
中科院分区：
文献类型：
--
作者：
Demidov, VV;Kuhn, H;Frank-Kameneteskii, MD
肽核酸 (PNA) 是一类合成的核酸聚酰胺模拟物，可提供多种应用。嘧啶双-PNA 可用于合理设计新颖的互锁 DNA 纳米结构、耳环标记，代表锁定的拟轮烷或锁定的索烷。这些结构是通过 DNA 连接酶介导的双链 DNA 与指定 DNA 位点处的环状寡核苷酸标签的连接而创建的。通过形成由一对双-PNA 开启子和探针寡核苷酸组成的 PD 环来进行组装。开启子局部暴露双链 DNA 的两条链之一，用于与探针杂交，其末端与置换的 DNA 链互补。杂交后，它们并列，随后可以通过 DNA 连接酶连接。结果，在 DNA 双螺旋上的精确位置形成真正的拓扑连接，产生锁定的耳环状标签。 DNA 拓扑标记既可以在溶液中进行，对于较长的模板，也可以在琼脂糖凝胶塞内进行。因此，通过DNA支架上各种配体的精确空间定位，利用杂交信号的滚环放大进行高度定位的DNA检测以及利用DNA双链体进行有效的显微操作成为可能。 (C) 2001 年学术出版社。
Peptide nucleic acids (PNAs) are a family of synthetic polyamide mimics of nucleic acids that offer a variety of applications. Pyrimidine bis-PNAs can be used for rational design of novel interlocked DNA nanostructures, earring labels, representing locked pseudorotaxanes or locked catenanes. These structures are created through DNA ligase-mediated catenation of duplex DNA with a circularized oligonucleotide tag at a designated DNA site. The assembly is performed via formation of the PD-loop consisting of a pair of bis-PNA openers and the probe oligonucleotide. The openers locally expose one of the two strands of duplex DNA for hybridizing the probe, whose termini are complementary to the displaced DNA strand. After hybridization, they are in juxtaposition and can subsequently be linked by DNA ligase. As a result, a true topological link forms at a precise position on the DNA double helix yielding locked, earring-like label. DNA topological labeling can be done both in solution and, for longer templates, within the agarose gel plug. Accordingly, highly localized DNA detection with rolling circle amplification of hybridization signal and effective micromanipulations with DNA duplexes become possible through precise spatial positioning of various ligands on the DNA scaffold. (C) 2001 Academic Press.