期刊
BIOSENSORS & BIOELECTRONICS
卷 26, 期 2, 页码 491-496出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2010.07.067
关键词
Quantum dot; Molecular beacon; beta-Lactamase; Escherichia coli; Fluorescence in situ hybridization (FISH)
类别
资金
- National Key Scientific Program (973)-Nanoscience and Nanotechnology [2006CB933100, 2011CB933600]
- NSFC [20621502, 20921062]
- National Natural Science Foundation of China [20833006, 0677044]
- Ministry of Public Health [2009ZX10004-107, 2008ZX10004-004]
Quantum dots (QDs) are inorganic fluorescent nanocrystals with excellent properties such as tunable emission spectra and photo-bleaching resistance compared with organic dyes, which make them appropriate for applications in molecular beacons. In this work, quantum dot-based molecular beacons (QD-based MBs) were fabricated to specifically detect beta-lactamase genes located in pUC18 which were responsible for antibiotic resistance in bacteria Escherichia coli (E. coli) DH5 alpha. QD-based MBs were constructed by conjugating mercaptoacetic acid-quantum dots (MAA-QDs) with black hole quencher 2 (BHQ2) labeled thiol DNA vial metal-thiol bonds. Two types of molecular beacons, double-strands beacons and hairpin beacons, were observed in product characterization by gel electrophoresis. Using QD-based MBs, one-step FISH in tiny bacteria DH5a was realized for the first time. QD-based MBs retained their bioactivity when hybridizing with complementary target DNA, which showed excellent advantages of eliminating background noise caused by adsorption of non-specific bioprobes and achieving clearer focus of genes in plasmids pUC18, and capability of bacterial cell penetration and signal specificity in one-step in situ hybridization. (C) 2010 Elsevier B.V. All rights reserved.
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