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DNA电子显微镜检查

DNA electron microscopy.

作者信息

Brack C

出版信息

CRC Crit Rev Biochem. 1981;10(2):113-69. doi: 10.3109/10409238109114551.

DOI:10.3109/10409238109114551
PMID:6163590
Abstract

In recent years DNA electron microscopy has become a tool of increasing interest in the fields of molecular genetics and molecular and cell biology. Together with the development of in vitro recombination and DNA cloning, new electron microscope techniques have been developed with the aim of studying the structural and functional organization of genetic material. The most important methods are based on nucleic acid hybridizations: DNA-DNA hybridization (heteroduplex, D-loop), RNA-DNA hybridization (R-loop), or combinations of both (R-hybrid). They allow both qualitative and quantitative analysis of gene organization, position and extension of homology regions, and characterization of transcription. The reproducibility and resolution of these methods make it possible to map a specific DNA region within 50 to 100 nucleotides. Therefore they have become a prerequisite for determining regions of interest for subsequent nucleotide sequencing. Special methods have been developed also for the analysis of protein-DNA interaction: e.g., direct visualization of specific protein-DNA complexes (enzymes, regulatory proteins), and analysis of structures with higher complexity (chromatin, transcription complexes).

摘要

近年来,DNA电子显微镜已成为分子遗传学、分子与细胞生物学领域中越来越受关注的一种工具。随着体外重组和DNA克隆技术的发展,人们开发了新的电子显微镜技术,旨在研究遗传物质的结构和功能组织。最重要的方法基于核酸杂交:DNA-DNA杂交(异源双链、D环)、RNA-DNA杂交(R环)或两者的组合(R杂交)。它们能够对基因组织、同源区域的位置和延伸进行定性和定量分析,并对转录进行表征。这些方法的可重复性和分辨率使得在50至100个核苷酸范围内绘制特定DNA区域成为可能。因此,它们已成为确定后续核苷酸测序感兴趣区域的先决条件。还开发了用于分析蛋白质-DNA相互作用的特殊方法:例如,直接观察特定的蛋白质-DNA复合物(酶、调节蛋白),以及分析具有更高复杂性的结构(染色质、转录复合物)。

相似文献

1
DNA electron microscopy.DNA电子显微镜检查
CRC Crit Rev Biochem. 1981;10(2):113-69. doi: 10.3109/10409238109114551.
2
Electron microscopy of nucleic acid.
Dan Med Bull. 1988 Feb;35(1):33-48.
3
Visual biochemistry: new insight into structure and function of the genome.
Methods Biochem Anal. 1982;28:201-65. doi: 10.1002/9780470110485.ch5.
4
Electron microscopic approaches to the study of bacterial DNA organization.
Microbiol Sci. 1988 Mar;5(3):68-73.
5
A new method for mapping nucleic acid sequence homology by electron microscopy.一种通过电子显微镜绘制核酸序列同源性图谱的新方法。
Nucleic Acids Res. 1975 Apr;2(4):509-20. doi: 10.1093/nar/2.4.509.
6
Cloning and delimiting one chloroplast DNA replicative origin of Chlamydomonas.衣藻叶绿体DNA一个复制起点的克隆与界定
Nucleic Acids Res. 1984 May 11;12(9):3857-72. doi: 10.1093/nar/12.9.3857.
7
Determination of cellular RNA concentrations by electron microscopy of R loop-containing DNA.通过含R环DNA的电子显微镜测定细胞RNA浓度。
Proc Natl Acad Sci U S A. 1981 May;78(5):2820-4. doi: 10.1073/pnas.78.5.2820.
8
Hybridization techniques.杂交技术。
Lab Res Methods Biol Med. 1982;5:185-229.
9
Electron microscopic visualization of nucleic acids and of their complexes with proteins.核酸及其与蛋白质复合物的电子显微镜观察。
Annu Rev Biochem. 1979;48:649-79. doi: 10.1146/annurev.bi.48.070179.003245.
10
Kinetics of spontaneous displacement of RNA from heteroduplexes by DNA.DNA介导RNA从异源双链体中自发置换的动力学
Nucleic Acids Res. 1996 Aug 15;24(16):3246-52. doi: 10.1093/nar/24.16.3246.

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