Quantum-dot-labeled DNA probes for fluorescence in situ hybridization (FISH) in the microorganism Escherichia coli.
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.
Wu
Sheng-Mei
SM
College of Chemistry and Molecular Sciences and State Key Laboratory of Virology, Wuhan University, Wuhan 430072, P. R. China.
Zhao
Xiang
X
Zhang
Zhi-Ling
ZL
Xie
Hai-Yan
HY
Tian
Zhi-Quan
ZQ
Peng
Jun
J
Lu
Zhe-Xue
ZX
Pang
Dai-Wen
DW
Xie
Zhi-Xiong
ZX
D015342:DNA Probes / Q000737:chemistry*; D004277:DNA, Single-Stranded / Q000737:chemistry; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry; D017404:In Situ Hybridization, Fluorescence / Q000295:instrumentation* / Q000379:methods; D008856:Microscopy, Fluorescence; D008956:Models, Chemical; D036103:Nanotechnology / Q000295:instrumentation* / Q000379:methods*; D009693:Nucleic Acid Hybridization; D010957:Plasmids / Q000378:metabolism; D045663:Quantum Dots; D012680:Sensitivity and Specificity; D013997:Time Factors
D015342:DNA Probes; D004277:DNA, Single-Stranded; D005456:Fluorescent Dyes