Fluorescence in situ hybridization (FISH) on maize metaphase chromosomes with quantum dot-labeled DNA conjugates.
Semiconductor nanocrystals, also called quantum dots (QDs), are novel inorganic fluorophores which are brighter and more photostable than organic fluorophores. In the present study, highly dispersive QD-labeled oligonucleotide (TAG)(8) (QD-deoxyribonucleic acid [DNA]) conjugates were constructed via the metal-thiol bond, which can be used as fluorescence in situ hybridization (FISH) probes. FISH analysis of maize metaphase chromosomes using the QD-DNA probes showed that the probes could penetrate maize chromosomes and nuclei and solely hybridized to complementary target DNAs. Compared with the conventional organic dyes such as Cy3 and fluorescein isothiocyanate, this class of luminescent labels bound with oligonucleotides is brighter and more stable against photobleaching on the chromosomes after FISH. These results suggest that QD fluorophores may be a more stable and useful fluorescent label for FISH applications in plant chromosome mapping considering their size-tunable luminescence spectra.
Ma
Lu
L
Key Laboratory of MOE for Plant Developmental Biology, College of Life Sciences, Wuhan University, Wuhan 430072, People's Republic of China.
Wu
Sheng-Mei
SM
Huang
Jing
J
Ding
Yi
Y
Pang
Dai-Wen
DW
Li
Lijia
L
D019921:Biotinylation; D002232:Carbocyanines / Q000494:pharmacology; D002467:Cell Nucleus / Q000378:metabolism; D002875:Chromosomes / Q000648:ultrastructure*; D004247:DNA / Q000737:chemistry; D015342:DNA Probes / Q000494:pharmacology; D016650:Fluorescein-5-isothiocyanate / Q000494:pharmacology; D017343:Genes, Plant; D006824:Hybridization, Genetic; D017404:In Situ Hybridization, Fluorescence; D008677:Metaphase; D009841:Oligonucleotides / Q000737:chemistry; D045663:Quantum Dots; D013056:Spectrophotometry, Ultraviolet; D003313:Zea mays / Q000235:genetics*
D002232:Carbocyanines; D015342:DNA Probes; D009841:Oligonucleotides; C089187:cyanine dye 3; D004247:DNA; D016650:Fluorescein-5-isothiocyanate