This work develops a fluorescence approach for sensitive detection of DNA methyltransferase activity based on endonuclease and rolling circle amplification (RCA) technique. In the presence of DNA adenine methylation...This work develops a fluorescence approach for sensitive detection of DNA methyltransferase activity based on endonuclease and rolling circle amplification (RCA) technique. In the presence of DNA adenine methylation (Dam) MTase, the methylation-responsive sequence of hairpin probe is methylated and cleaved by the methylation-sensitive restriction endonuclease Dpn 1. The products cleaved by restriction endonuclease Dpn I then function as a signal primer to initiate RCA reaction by hybridizing with the circular DNA template. Each RCA product containing thousands of repeated sequences might hybridize with a large number of molecular beacons (detection probes), resulting in an enhanced fluorescence signal. In the absence of Dam MTase, neither methylation/cleavage nor RCA reaction can be initiated and no fluorescence signal is observed. The proposed method exhibits a dynamic range from 0.5 U/mL to 30 U/mL and a detection limit of 0.18 U/mL. This method can be used for the screening of antimicrobial drugs and has a great potential to be further applied in early clinical diagnosis.展开更多
A novel method for the detection of PDGF-BB has been developed using double-strand DNA-copper nanoparticles (dsDNA-CuNPs) as fluorescent markers. This assay relies on the premise that the aptamer- based probe underg...A novel method for the detection of PDGF-BB has been developed using double-strand DNA-copper nanoparticles (dsDNA-CuNPs) as fluorescent markers. This assay relies on the premise that the aptamer- based probe undergoes a conformational change upon binding with target protein, and subsequently triggers polymerization reaction to generate dsDNA. Then, the resultant dsDNA can be used as a template for the formation of CuNPs with high fluorescence. Under the optimized conditions, the proposed assay allowed sensitive and selective detection of PDGF-BB with a detection limit of 4 nmol/L. This possibly makes it an attractive platform for the detection of a variety of biomolecules whose aptamers undergo similar conformational change.展开更多
A one-step and facile strategy toward the construction of multicolor polymers via supramolecular selfassembly was proposed.Multicolor polymers were simply prepared by the self-assembly of adamantane-labeled fluorescei...A one-step and facile strategy toward the construction of multicolor polymers via supramolecular selfassembly was proposed.Multicolor polymers were simply prepared by the self-assembly of adamantane-labeled fluorescein,adamantane-labeled rhodamine B and β-cyclodextrin polymers via host-guest interaction between β-cyclodextrin and adamantane.Multicolor polymers showed several interesting properties:multiple emission signatures by a single wavelength excitation,easy tunability,intense fluorescence,high photostablility.In addition,the self-assembly approach implied a facile and flexible strategy for constructing functionalized materials,such as multicolor materials for biological labeling and imaging,and sensing materials for the detection of physiological parameters.展开更多
基金supported by the National Natural Science Foundation of China(Nos.21190044 and 21175035)National Basic Research Program(No.2011CB911002)International Science&Technology operation Program of China(No.2010DFB30300)
文摘This work develops a fluorescence approach for sensitive detection of DNA methyltransferase activity based on endonuclease and rolling circle amplification (RCA) technique. In the presence of DNA adenine methylation (Dam) MTase, the methylation-responsive sequence of hairpin probe is methylated and cleaved by the methylation-sensitive restriction endonuclease Dpn 1. The products cleaved by restriction endonuclease Dpn I then function as a signal primer to initiate RCA reaction by hybridizing with the circular DNA template. Each RCA product containing thousands of repeated sequences might hybridize with a large number of molecular beacons (detection probes), resulting in an enhanced fluorescence signal. In the absence of Dam MTase, neither methylation/cleavage nor RCA reaction can be initiated and no fluorescence signal is observed. The proposed method exhibits a dynamic range from 0.5 U/mL to 30 U/mL and a detection limit of 0.18 U/mL. This method can be used for the screening of antimicrobial drugs and has a great potential to be further applied in early clinical diagnosis.
基金supported by the National Natural Science Foundation of China(Nos.21190041,21190044,21175035)National Basic Research Program(No.2011CB911002)Interna-tional Science&Technology operation Program of China(No.2010DFB30300)
文摘A novel method for the detection of PDGF-BB has been developed using double-strand DNA-copper nanoparticles (dsDNA-CuNPs) as fluorescent markers. This assay relies on the premise that the aptamer- based probe undergoes a conformational change upon binding with target protein, and subsequently triggers polymerization reaction to generate dsDNA. Then, the resultant dsDNA can be used as a template for the formation of CuNPs with high fluorescence. Under the optimized conditions, the proposed assay allowed sensitive and selective detection of PDGF-BB with a detection limit of 4 nmol/L. This possibly makes it an attractive platform for the detection of a variety of biomolecules whose aptamers undergo similar conformational change.
基金supported by the National Natural Science Foundation of China(Nos.21190044,21175035)National Basic Research Program(No.2011CB911002)International Science & Technology operation Program of China(No.2010DFB30300)
文摘A one-step and facile strategy toward the construction of multicolor polymers via supramolecular selfassembly was proposed.Multicolor polymers were simply prepared by the self-assembly of adamantane-labeled fluorescein,adamantane-labeled rhodamine B and β-cyclodextrin polymers via host-guest interaction between β-cyclodextrin and adamantane.Multicolor polymers showed several interesting properties:multiple emission signatures by a single wavelength excitation,easy tunability,intense fluorescence,high photostablility.In addition,the self-assembly approach implied a facile and flexible strategy for constructing functionalized materials,such as multicolor materials for biological labeling and imaging,and sensing materials for the detection of physiological parameters.