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用于微生物大肠杆菌荧光原位杂交(FISH)的量子点标记DNA探针。

Quantum-dot-labeled DNA probes for fluorescence in situ hybridization (FISH) in the microorganism Escherichia coli.

作者信息

Wu Sheng-Mei, Zhao Xiang, Zhang Zhi-Ling, Xie Hai-Yan, Tian Zhi-Quan, Peng Jun, Lu Zhe-Xue, Pang Dai-Wen, Xie Zhi-Xiong

机构信息

College of Chemistry and Molecular Sciences and State Key Laboratory of Virology, Wuhan University, Wuhan 430072, P. R. China.

出版信息

Chemphyschem. 2006 May 12;7(5):1062-7. doi: 10.1002/cphc.200500608.

Abstract

Semiconductor quantum dots (QDs) as a kind of nonisotopic biological labeling material have many unique fluorescent properties relative to conventional organic dyes and fluorescent proteins, such as composition- and size-dependent absorption and emission, a broad absorption spectrum, photostability, and single-dot sensitivity. These properties make them a promising stable and sensitive label, which can be used for long-term fluorescent tracking and subcellular location of genes and proteins. Here, a simple approach for the construction of QD-labeled DNA probes was developed by attaching thiol-ssDNA to QDs via a metal-thiol bond. The as-prepared QD-labeled DNA probes had high dispersivity, bioactivity, and specificity for hybridization. Based on such a kind of probe with a sequence complementary to multiple clone sites in plasmid pUC18, fluorescence in situ hybridization of the tiny bacterium Escherichia coli has been realized for the first time.

摘要

半导体量子点(QDs)作为一种非同位素生物标记材料,相对于传统有机染料和荧光蛋白具有许多独特的荧光特性,如组成和尺寸依赖性吸收与发射、宽吸收光谱、光稳定性和单量子点灵敏度。这些特性使其成为一种有前景的稳定且灵敏的标记物,可用于基因和蛋白质的长期荧光追踪及亚细胞定位。在此,通过金属硫醇键将巯基化单链DNA连接到量子点上,开发了一种构建量子点标记DNA探针的简单方法。所制备的量子点标记DNA探针具有高分散性、生物活性和杂交特异性。基于这种与质粒pUC18中多个克隆位点序列互补的探针,首次实现了对微小细菌大肠杆菌的荧光原位杂交。

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