Analytical Methods
·
2019
·Vol. 11
·期 28
·3584-3589
·
IF 2.378
Yanqiong Lai,Xia Teng,Yanli Zhang,Hongbin Wang,Pengfei Pang,Chun Yang,Colin J. Barrow,Wenrong Yang
In this study, we developed a simple, sensitive, low-cost and label-free method to detect rutin by using double-stranded DNA-templated copper nanoclusters (dsDNA-CuNCs) as a fluorescent probe. The dsDNA-template can facilitate the formation of highly fluorescent CuNCs with a maximum emission wavelength of 575 nm. In the dsDNA-CuNC fluorescent probe, rutin was intercalated into the structure of dsDNA via hydrogen bonding and electrostatic interactions. The fluorescence of dsDNA-CuNCs was quenched considerably due to the electron transfer from CuNCs to rutin. This decrease in the fluorescence intensity of dsDNA-CuNCs can be used to monitor the concentration of rutin from 0.6 to 400 μM with a detection limit of 0.12 μM (S/N = 3). Compared with the previous reports, this method does not require any complex DNA sequence design, fluorescent dye label and sophisticated experimental techniques. Moreover, the proposed sensing system could be applicable for the detection of rutin in pharmaceutical samples.
研究领域:
生物化学
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分析化学
·
药物分析
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DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2021
·
IF 7.46
Dawei Li,Shen Ling,Hongsong Wu,Zhaoqi Yang,Bei Lv
In addition to its application in genome editing, the CRISPR/Cas system provides a new molecular tool for the construction of biosensor platforms due to its high target sequence specificity and programmability . To exploit the potential of the CRISPR/Cas system for detecting non-nucleic-acid targets, two ultrasensitive biosensors (sensor-ss and sensor-ds) consisting of allosteric aptamer probes integrated with the CRISPR/Cas12a system were designed and constructed for tobramycin detection. Both sensors utilized hairpin DNA containing a tobramycin aptamer sequence as the target recognition probe, which can respond to target binding and further produce signal transduction sequences. However, there are two differences between the sensors. First, in sensor-ss, single-stranded DNA was set to be the activator to trigger cutting activity ( trans -cleavage) of CRISPR/Cas12a, leading to cleavage of the FQ-reporter (a short single-stranded DNA labeled with a fluorophore and quencher at each end) and increasing the fluorescent signal. However, double-stranded DNA containing a protospacer adjacent motif (PAM) site was produced in sensor-ds as the signal transduction sequence, which was recognized by CRISPR/Cas12a, resulting in the cleavage of the FQ-reporter. Second, sensor-ss involved a strand displacement amplification (SDA) reaction as the signal enhancement step, while no DNA amplification procedure was designed in sensor-ds. Through DNA sequence and reaction condition optimization, sensor-ds showed a linear relationship between the fluorescence response and tobramycin concentrations ranging from 10 to 300 pM with a limit of detection (LOD) as low as 3.719 pM. Although sensor-ss exhibited higher sensitivity (LOD = 1.542 pM) than sensor-ds, rapidly reaching the fluorescent signal saturation platform led to a narrow linear response range (5–30 pM). Moreover, our CRISPR/Cas12a-based assay can realize on-site detection by the naked eye readout under UV light. The constructed sensor-ds was also used to detect tobramycin in milk and lake water samples, and the results indicated that the detection system was reliable. The design strategy reported in this study could shed new light on CRISPR/Cas12a-based biosensor construction for small molecule detection.
研究领域:
分子生物学
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分析化学
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医学检测
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DOI
高影响因子
ACS Omega
·
2019
·
IF 2.584
Yu Ding,Xianming Li,Xinfeng Zhang,Feng Li,Xiandeng Hou,Peng Wu
Exonuclease III (Exo III) is an important enzymatic tool that is being widely used in molecular biology, biotechnology, and bioassay development. Exo III prefers to cleave double-stranded DNA (dsDNA) with blunt and recessed 3′-termini rather than their protruding counterpart. While it has been accepted that a short 3′-overhang (e.g., >4 nt) is necessary to protect a dsDNA from Exo III cleavage, critical roles of the length and sequence of this 3′-overhang remain unexplored. Herein, we develop a novel light-induced colorimetric assay allowing the systematic probe of the sequence selectivity of Exo III in a rapid and high-throughput manner. Our finding that Exo III is highly specific to 3′-overhang in terms of both length and sequence will be valuable for guiding the design of bioassays and DNA manipulating tools mediated by Exo III.
研究领域:
分子生物学
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生物技术
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酶学研究
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DOI
PubMed
高影响因子
Polymers
·
2022
·Vol. 14
·期 19
·4133
·
IF 4.967
Lianming Zhang,Kui Luo,Jingxia Gao,Jianping Li
A new chiral molecularly imprinted polymer (MIP) sensor with dual recognition ability was developed for the highly selective separation of enantiomers with toxic side effects in drugs. The sensor contains double-stranded deoxyribonucleic acid (dsDNA) as the element that immobilizes the chiral molecular conformation: the dsDNA enables the imprinted cavities to match the three-dimensional structure and functional groups from the chiral molecule. By embedding the spatial orientation of dsDNA in MIPs, one can accurately capture and immobilize the molecular conformation, eliminating the influence of interfering analogues. Herein, L-penicillamine (L-Pen) was selected as the chiral template molecule and embedded into dsDNA to form dsDNA-L-Pen complex, which was then embedded into the MIPs by electropolymerization. After elution, the stereo-selective imprinted cavities were obtained. The ATATATATATAT-TATATATATATA base sequence showed a high affinity for the embedded L-Pen, which endowed the imprinted cavities with a larger number of sites and improved the selectivity toward Pen enantiomers. Under the optimal working conditions, the current response of the MIP/dsDNA sensor exhibited a positive linear relationship with the logarithm of the L-Pen concentration in the range of 3.0 × 10−16to 3.0 × 10−13mol/L, and the detection limit was 2.48 × 10−16mol/L. After the introduction of dsDNA into the MIP, the selectivity of the sensor toward D-Pen increased by 6.4 times, and the sensor was successfully applied in the analysis of L-Pen in penicillamine tablets.Keywords:molecularly imprint;sensor;dsDNA;chiral;L-Pen
研究领域:
分析化学
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生物医学工程
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2017
·
IF 6.32
Jinyang Chen,Xinghu Ji,Zhike He
On the basis of an interesting experimental phenomenon, a novel and smart composite reagent consisting of double-stranded DNA-templated copper nanoparticles (dsDNA–CuNPs) and DNA intercalator (SYBR Green I) was developed and exploited for hydrogen peroxide (H2O2) detection as well as oxidase-based biosensing. The study found that, within the composite reagent, the small molecule SYBR Green I was easily adsorbed on the surfaces of CuNPs, instead of intercalating into the dsDNA. So the composite reagent only exhibited the red fluorescence generated from dsDNA–CuNPs. However, when the solution of H2O2 was added into the composite reagent, the CuNPs were deconstructed and their fluorescence was quenched; meanwhile, the inhibition of SYBR Green I binding with dsDNA was eliminated. As a result, the mixed solution of the composite reagent with H2O2 exhibited green fluorescence generated from the intercalation of SYBR Green I into dsDNA. Since H2O2 is an important molecule and involved in various research fields, this developed composite reagent could be employed for many applications in biological analysis. As a proof-of-application demonstration, the sensitive detection of glucose was conducted. Moreover, the method was also extended to the detection of other biomolecules, such as cholesterol and horseradish peroxidase, which indicated the broad applications of the proposed sensing strategy in biomedical analysis.
研究领域:
生物化学
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分析化学
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生物医学
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DOI
PubMed
高影响因子
DYES AND PIGMENTS
·
2018
·
IF 3.767
Wei Long,Yu-Jing Lu,Kun Zhang,Xuan-He Huang,Jin-Qiang Hou,Sen-Yuan Cai,Ying Li,Xian Du,Leonard G. Luyt,Wing-Leung Wong,Cheuk-Fai Chow
New fluorescent dyes derivatized on a classical thiazole orange framework are able to show unexpectedly strong interaction signal and unusual binding selectivity with different structures of nucleic acid , particularly when bound with double-stranded DNA or G-quadruplex DNA. The present study reveals that these small binding ligands simply bearing an additional amino side group on its parent molecule of thiazole orange have almost no background fluorescence in solution. Conversely, they are able to produce an extremely strong yellow emission signal upon interaction with targeting nucleic acids in live cells. The induced fluorescence intensity was approximately 10–15 times stronger than that of thiazole orange. The dyes are therefore excellent fluorescent stains for bio-sensing and bio-imaging applications. It is particularly suitable for fluorescence microscopy experiments requiring very low working concentration (0.25 μM or less) targeting nucleic acids. This was demonstrated using pu27 G-quadruplex DNA, which has a low limit of detection (LOD = 3–4 nM) while thiazole orange was much higher (LOD = 48.7 nM) under the same conditions. In addition, it was found that structural modification on the quinolinium scaffold of thiazole orange was less effective than modification on the benzothiazole moiety. The findings of the present study provide important information for structural advancement of small molecules based on the widely used thiazole orange skeleton, resulting in analogues that are able to achieve higher sensitivity and selectivity for targeting at a specific class of nucleic acids. Computational docking studies were also conducted to illustrate the interaction behaviors of the dyes with different DNA structures.
研究领域:
生物化学
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分子生物学
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分析化学
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高影响因子
JOURNAL OF POLYMER SCIENCE PART B-POLYMER PHYSICS
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2017
·Vol. 56
·期 6
·501-508
·
IF 2.838
Sha Wu,Xiangjun Gong,Geoffrey I. N. Waterhouse,Guangzhao Zhang
The effect of monovalent and divalent cations on the ssDNA to dsDNA transformation was systematically examined. For salts containing monovalent cations (LiCl, NaC1, KCl, CsCl), the conversion of ssDNA to dsDNA increased with ionic strength up to a value of I = 0.01 M and then plateaued, confirming that all four of the monovalent cations behaved similarly and promoted the formation of dsDNA. The monovalent cation type influenced the equilibrium constant for the conversion of ssDNA to dsDNA, indicating a degree of ion-specificity in dsDNA formation. In the case of salts containing divalent cations (e.g., MgCl 2 ), the conversion of ssDNA to dsDNA also increased with increasing ionic strength, though the plateau region was reached at a much lower ionic strength ( I = 5.0 × 10 −4 M), which can be attributed to the higher electrostatic screening efficiency of Mg 2+ cations and thus their superior ability to link DNA chains. © 2017 Wiley Periodicals, Inc. J. Polym. Sci., Part B: Polym. Phys. 2018 , 56 , 501–508
研究领域:
生物化学
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分子生物学
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DOI
高影响因子
BIOELECTROCHEMISTRY
·
2024
·
IF 4.8
Longjian Huang,Wenzhao Zhang,Mingxuan Liu,Yuanxun Gong,Qianli Tang,Kaihua Wang,Xianjiu Liao,Kai Zhang,Jihua Wei
We present an innovative biosensor designed for the precise identification of Escherichia coli ( E.coli ), a predominant pathogen responsible for gastrointestinal infections. E.coli is prevalent in environments characterized by substandard water quality and can lead to severe diarrhea, especially in hospital settings. The device employs entropy-driven reactions to synthesize copious amounts of double-stranded DNA (dsDNA), which, upon binding with crRNA, triggers the CRISPR/Cas12a system’s cleavage mechanism. This process results in the separation of a ferrocene (Fc)-tagged DNA strand from the electrode, enhancing the electrochemical signal for E.coli ’s rapid and accurate detection. Our tests confirm the biosensor’s ability to quantify E.coli across a dynamic range from 100 to 10 million CFU/mL, achieving a detection threshold of just over 5 CFU/mL. The development of this electrochemical biosensor highlights its exceptional selectivity, high sensitivity, and user-friendly interface for E.coli detection. It stands as a significant step forward in pathogen detection technology, promising new directions for identifying various bacterial infections through the CRISPR/Cas mechanism.
研究领域:
分子生物学
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病原体检测
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生物传感技术
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2018
·
IF 5.667
Jing Wang,Tingting Li,Haigang Li,Gongke Li,Shixin Wu,Liansheng Ling
AuNPs-DNA probes remain a challenge to recognize the double-stranded DNA (dsDNA) from routine PCR product despite its success in the recognition of the single-stranded (ssDNA) DNA of asymmetry-PCR product. Here, a universal colorimetric PCR biosensor is established with oligonucleotide modified AuNPs by recognizing the dsDNA of PCR product based on triplex DNA formation with the assistance of a triplex DNA intercalator Ru(phen) 2 dppx 2+ . The strategy was applied to detect different length dsDNA, including short dsDNA 1 (49 bp) and long dsDNA 2 (321 bp), both of which had the good linear relationship between the absorbance and the logarithm of the concentration of the targets. Moreover, the limit of detection (LOD) for short and long length dsDNA was 0.5 pM and 0.05 fg, lower than those of the previously reported methods, respectively. This strategy also displayed high selectivity against dsDNA 1 and dsDNA 2. These results demonstrate a potential strategy to detect all length of dsDNA, providing new opportunities in the life science, diagnosis, and medicine.
研究领域:
生物化学
·
分子诊断
·
纳米医学
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产品关联
DOI
高影响因子
PARTICLE & PARTICLE SYSTEMS CHARACTERIZATION
·
2023
·Vol. 40
·期 9
·2300041
·
IF 2.7
Wenhui Bao,Jun Ai
In this work, silver nanoparticles are synthesized using a simple and sensitive method by using double-stranded DNA (dsDNA-Ag NPs) as a template. The prepared dsDNA-Ag NPs are characterized by fluorescence spectroscopy analysis, X-ray photoelectron spectroscopy analysis, and transmission electron microscopy analysis. The excitation wavelength of the prepared silver nanoparticles is 295 nm, the emission wavelength is 377 nm, the average particle size is 11.2 nm, and the dispersion is uniform with pleasurable stability. The nanomaterials are used as fluorescent probes to detect glutathione (GSH). After adding glutathione to the dsDNA-Ag NPs fluorescent probes, the fluorescence of dsDNA-Ag NPs is burst due to electron transfer and S Ag bond generation, and the linear range of detection concentration is 0–90 m m with a detection limit of 0.37 m m .
研究领域:
生物化学
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纳米生物学
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分析化学
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DOI
高影响因子
ANALYTICA CHIMICA ACTA
·
2015
·
IF 4.513
Xiaoyun Lin,Yongnian Ni,Serge Kokot
An electrochemical biosensor capable of indirect detection of DNA damage induced by any one of the three endocrine-disrupting compounds (EDCs) – bisphenol A (BPA), 4-nonylphenol (NP) and 4- t -octylphenol (OP), has been researched and developed. The methylene blue (MB) dye was used as the redox indicator. The glassy carbon electrode (GCE) was modified by the assembled dsDNA/graphene oxide-chitosan/gold nano-particles to produce a dsDNA/GO-CS/AuNPs/GCE sensor. It was characterized with the use of electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV) and scanning electron microscopy (SEM) techniques. The loading/release of the MB dye by the dsDNA/GO-CS/AuNPs film was investigated, and the results showed that the process was reversible. Based on this, the sensor was used to measure the difference between the loading capabilities of intact and damaged dsDNA in the films. The sensor was then successfully applied to detect DNA damage electrochemically. The differential pulse voltammetry (DPV) peak current ratio for MB, observed before and after DNA damage, increased linearly in the presence the BPA, NP or OP compounds; the treatment range was 10–60 min, and the respective damage rates were 0.0069, 0.0044 and 0.0031 min −1 , respectively. These results were confirmed by the binding constants: 2.09 × 10 6 M −1 (BPA-DNA), 1.28 × 10 6 M −1 (NP-DNA) and 9.33 × 10 5 M −1 (OP-DNA), all of which were obtained with the use of differential pulse stripping voltammetry (DPSV).
研究领域:
生物化学
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环境科学
·
毒理学
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产品关联
DOI
PubMed
高影响因子
CHINESE JOURNAL OF ANALYTICAL CHEMISTRY
·
2018
·
IF 0.824
Lian-Ming ZHANG,Dong-You ZHANG,Ying ZENG,Jian-Ping LI
A novel strategy based on DNA induced conformation change was developed by rendering the imprinting cavity with refined structure in order to further improve the selectivity of molecular imprinting membrane. The double-stranded DNA (dsDNA) were first self-assembled on the surface of the gold electrode, then cimaterol (CIM) molecule, taken as the representative target, would bind with the groove of dsDNA to form complex template, together with which the molecular imprinting polymers (MIPs) was prepared by electropolymerization in the monomer solution. After the elution of CIM, the obtained MIPs sensor had a remarkably high selectivity toward CIM. The combination process of CIM and DNA was studied by UV-Vis spectroscopy and was found to follow the groove combination model. The optimal conditions of the system were optimized by electrochemical method. The results showed that the sensor had the best recognition ability toward CIM when the elution time was 35 min by ethanol-acetic acid (8:1, V/V ) as an eluent and rebound time was 45 min. This sensor exhibited a linear detection range of 1.0 × 10 −13 −1.0 × 10 −10 M with a detection limit of 3.2 × 10 −14 M. The sensor was successfully applied for detection of CIM in pork samples.
研究领域:
生物化学
·
分析化学
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产品关联
DOI
高影响因子
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