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