Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.2.1.17 (lysozyme)
 21,489 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A new strategy for quantitatively detecting micrococcal nuclease (MNase) is proposed using electrostatic interaction-based fluorescence resonance energy transfer (FRET) between positively charged QDs and negatively charged dye-labeled single-stranded DNA (dye-ssDNA). Herein, we have made our attempt to develop a strategy where the variation of FRET efficiency is due to the change of the electrostatic interaction between QDs and the ssDNA that result from the cleavage of dye-ssDNA by a single-strand-specific nuclease. To demonstrate the feasibility of this design, positively charged QDs (lysozyme modified QDs, Lyz-QDs) are prepared as the energy donor, with the fluorescent dye 6-carboxy-X-rhodamine (ROX) that is labeled to ssDNA serving as the energy acceptor. The ROX-labeled probe ssDNA (ROX-ssDNA) is absorbed to the surface of the QDs through electrostatic interaction, which results in resonance energy transfer between the QDs and the dye. In the presence of MNase which cleaves the ROX-ssDNA into small fragments, the weakened interaction between QDs and the shortened ssDNA causes the decrease of FRET efficiency. At given amounts of donor and acceptor, the ratio of fluorescence intensity of QDs to ROX changes in a MNase concentration-dependent manner. Under optimized conditions, the ratio is linear with MNase concentration over the range of 8 x 10(-3) to 9.0 x 10(-2) unit mL(-1), with a limit of detection of 1.6 x 10(-3) unit mL(-1). This new detection strategy features straightforward design and easy operation, which is capable of expanding the application of the positively charged QDs-based FRET in DNA-related bioassays.
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PMID:A positively charged QDs-based FRET probe for micrococcal nuclease detection. 2067 36

In this article, we report a novel dual on/off thrombin fluorescence aptasensor by combining the energy driven target induced strand displacement reaction and a non-enzyme catalyst recycling DNA machine. Firstly, the specific binding of an aptamer strand and thrombin induce the release of a catalyst which was used as a DNA machine trigger. Subsequently, the catalyst as the trigger initiated the DNA machine through nucleic acid hybridization and branch migration of the DNA machine, resulting in the DNA substrate melting and re-hybridization. In such a working model, the DNA machine achieved cooperative control of the circular strand displacement reaction, realizing catalyst recycling and dual-amplification. The fluorescence signal change of FAM and ROX accumulation had a good linear relationship with the thrombin concentration in the range of 1 fM to 1 nM. On account of catalyst recycling and dual recognition, the detection limit for thrombin was determined to be as low as 0.45 fM (S/N = 3).This biosensor also showed good selectivity for thrombin without being affected by some other proteins, such as PSA, lysozyme etc. Moreover, this assay can be applied to the detection of thrombin in diluted human serum.
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PMID:An off/on thrombin activated energy driven molecular machine for sensitive detection of human thrombin via non-enzymatic catalyst recycling amplification. 3282 Feb 97