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本文引用的文献

1
Inhibition of gene expression by triple helix-directed DNA cross-linking at specific sites.通过在特定位点进行三链螺旋导向的DNA交联来抑制基因表达。
Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3501-5. doi: 10.1073/pnas.90.8.3501.
2
Electron microscopy visualization of oligonucleotide binding to duplex DNA via triplex formation.通过三链体形成对与双链DNA结合的寡核苷酸进行电子显微镜观察。
J Mol Biol. 1993 Mar 20;230(2):379-83. doi: 10.1006/jmbi.1993.1154.
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A possible family of B-like triple helix structures: comparison with the Arnott A-like triple helix.一种可能的B类三螺旋结构家族：与阿诺特A类三螺旋的比较。
Biochemistry. 1993 Mar 2;32(8):2098-103. doi: 10.1021/bi00059a030.
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Targeted mutagenesis of DNA using triple helix-forming oligonucleotides linked to psoralen.使用与补骨脂素相连的三链螺旋形成寡核苷酸对DNA进行靶向诱变。
Proc Natl Acad Sci U S A. 1993 Aug 15;90(16):7879-83. doi: 10.1073/pnas.90.16.7879.
5
Influence of fluctuations on DNA curvature. A comparison of flexible and static wedge models of intrinsically bent DNA.波动对DNA曲率的影响。固有弯曲DNA的柔性和静态楔形模型的比较。
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Oligonucleotide clamps arrest DNA synthesis on a single-stranded DNA target.寡核苷酸钳制可阻止单链DNA靶标上的DNA合成。
Proc Natl Acad Sci U S A. 1993 Nov 1;90(21):10013-7. doi: 10.1073/pnas.90.21.10013.
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Targeted mutagenesis of simian virus 40 DNA mediated by a triple helix-forming oligonucleotide.由三链螺旋形成寡核苷酸介导的猿猴病毒40 DNA的靶向诱变
J Virol. 1993 Dec;67(12):7324-31. doi: 10.1128/JVI.67.12.7324-7331.1993.
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DNA curvature influences the internal motions of supercoiled DNA.DNA弯曲影响超螺旋DNA的内部运动。
EMBO J. 1993 Nov;12(11):4407-12. doi: 10.1002/j.1460-2075.1993.tb06125.x.
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A parallel DNA triplex as a model for the intermediate in homologous recombination.作为同源重组中间体模型的平行DNA三链体
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Short-range DNA looping by the Xenopus HMG-box transcription factor, xUBF.非洲爪蟾HMG盒转录因子xUBF介导的短程DNA环化
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通过三链螺旋形成和补骨脂素交联对超螺旋DNA进行序列特异性标记。

Sequence-specific labeling of superhelical DNA by triple helix formation and psoralen crosslinking.

作者信息

Pfannschmidt C, Schaper A, Heim G, Jovin T M, Langowski J

机构信息

German Cancer Research Center, Heidelberg, Germany.

出版信息

Nucleic Acids Res. 1996 May 1;24(9):1702-9. doi: 10.1093/nar/24.9.1702.

DOI:10.1093/nar/24.9.1702
PMID:8649989
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC145834/
Abstract

Site-specific labeling of covalently closed circular DNA was achieved by using triple helix-forming oligonucleotides 10, 11 and 27 nt in length. The sequences consisted exclusively of pyrimidines (C and T) with a reactive psoralen at the 5'-end and a biotin at the 3'-end. The probes were directed to different target sites on the plasmids pUC18 (2686 bp), pUC18/4A (2799 bp) and pUC1 8/4A-H 1 (2530 bp). After triple helix formation at acid pH the oligonucleotides were photocrosslinked to the target DNAs via the psoralen moiety, endowing the covalent adduct with unconditional stability, e.g. under conditions unfavorable for preservation of the triplex, such as neutral pH. Complex formation was monitored after polyacrylamide gel electrophoresis by streptavidin-alkaline phosphatase (SAP)-induced chemiluminescence. The yield of triple helix increased with the molar ratio of oligonucleotide to target and the length of the probe sequence (27mer > 11mer). The covalent adduct DNA were visualized by scanning force microscopy (SFM) using avidin or streptavidin as protein tags for the biotin group on the oligonucleotide probes. We discuss the versatility of triple helix DNA complexes for studying the conformation of superhelical DNA.

摘要

通过使用长度为10、11和27个核苷酸的三链螺旋形成寡核苷酸实现了共价闭合环状DNA的位点特异性标记。这些序列完全由嘧啶（C和T）组成，在5'端带有一个反应性补骨脂素，在3'端带有一个生物素。这些探针靶向质粒pUC18（2686 bp）、pUC18/4A（2799 bp）和pUC18/4A-H1（2530 bp）上的不同靶位点。在酸性pH下形成三链螺旋后，寡核苷酸通过补骨脂素部分与靶DNA进行光交联，使共价加合物具有无条件稳定性，例如在不利于三链体保存的条件下，如中性pH。通过链霉亲和素-碱性磷酸酶（SAP）诱导的化学发光在聚丙烯酰胺凝胶电泳后监测复合物的形成。三链螺旋的产率随着寡核苷酸与靶标的摩尔比以及探针序列的长度增加而增加（27聚体>11聚体）。使用抗生物素蛋白或链霉亲和素作为寡核苷酸探针上生物素基团的蛋白质标签，通过扫描力显微镜（SFM）观察共价加合物DNA。我们讨论了三链螺旋DNA复合物在研究超螺旋DNA构象方面的多功能性。