BIOSENSORS & BIOELECTRONICS
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2013
·
IF 5.437
Lichun Zheng,Xiaoyan Li,Panpan Liu,Guofan Wu,Xiaoquan Lu,Xiuhui Liu
We present a novel strategy for simultaneous electrochemical detection of multiple DNA targets based on the use of different encoding metal ions as tags. The principle of this scheme is that metal ions bound to metallothionein (MT) molecules can be released down after hybridization with DNA targets and then be detected by stripping voltammetry . The novel detection probes, ssDNA/MT conjugates , covered with different metal ions were synthesized for the first time, then three encoding metal ions (Zn 2+ , Cd 2+ , and Pb 2+ ) were used to differentiate the signals of three virus DNA due to their well-defined anodic stripping peaks at −1.13 V (Zn), −0.78 V (Cd), and −0.52 V (Pb) at BiFE , respectively. The anodic peak currents increased linearly with the concentrations of DNA targets in the range from 0.1 nM to 10 nM with a detection limit of 33 pM. In addition, the one-base mismatched target was effectively discriminated from the complementary target. The described results demonstrated that this method possesses high sensitivity and selectivity for multi-target DNA assay and has great potential in applications for detection of even more targets in biological assays , particularly immunoassays .
研究领域:
生物化学
·
分子生物学
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2015
·
IF 6.409
Pei Liu,Dandan Wang,Yunlei Zhou,Haiyan Wang,Huanshun Yin,Shiyun Ai
DNA methyltransferase (MTase) has received a large amount of attention due to its catalyzation of DNA methylation in both eukaryotes and prokaryotes , which has a close relationship to cancer and bacterial diseases. Herein, a novel electrochemical strategy based on Dpn I digestion triggering the combination of poly adenine (polyA) DNA with a gold nanoparticles functioned glassy carbon electrode (AuNPs/GCE), is developed for the simple and efficient detection of DNA MTase and inhibitor screening. Only one methylene blue (MB)-labeled DNA hairpin probe and two enzymes are involved in this designed method. In the presence of Dam MTase, the hairpin probe can be methylated and then cleaved by the restriction endonuclease . Thus, a MB-labeled polyA signal-stranded DNA product is introduced to the surface of AuNPs/GCE through the effect between polyA and AuNPs, resulting in an obvious electrochemical signal. On the contrary, in the absence of Dam MTase, the DNA probe cannot be cleaved and a relatively small electrochemical response can be observed. As a result, the as-proposed biosensor offered an efficient way for Dam MTase activity monitoring with a low detection of 0.27 U/mL, a wide linear range and good stability. Additionally, this assay holds great potential for further application in real biological matrices and inhibitors screening, which is expected to be useful in disease diagnosis and drug discovery.
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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癌症研究
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微生物学
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DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
·
2020
·
IF 3.594
Guoli Xu,Danqun Huo,Jingzhou Hou,Chao Zhang,Yanan Zhao,Changjun Hou,Jing Bao,Xin Yao,Mei Yang
A novel electrochemical aptamer sensor platform for malathion detection was successfully constructed based on nanocomposite probe. The zirconium-based metal organic framework (MOF) was synthesized by hydrothermal method, and was coupled with complementary probe (CP) and carboxy-ferrocene (Fc-COOH) by covalent bonds to form CP-MOF-Fc nanocomposite probe. The complex probe hybridized with malathion aptamer on electrode by complementary base-pairing reactions. On account of the stronger affinity of the aptamer to target, CP-MOF-Fc was released into the buffer solution, causing a decrease in the DPV signal of Fc. Under optimized conditions, the prepared sensor showed a linear range of 25–850 ng L −1 toward malathion with a detection limit of 17.18 ng L −1 (S/N = 3) due to the high specific surface area of MOF , showing the excellent ability of signal amplification. In addition, it exhibited good repeatability and excellent selectivity. In the real sample experiment, satisfactory spike recovery results were obtained from 95.00 to 103.16%. The proposed strategy exhibited a rosy prospect in aptamer sensor design for pesticide testing.
研究领域:
分析化学
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电化学
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纳米技术
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环境科学
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产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2022
·
IF 9.221
Mei Wang,Hanfeng Cui,Nian Hong,Qingxia Shu,Xinru Wang,Yuping Hu,Guobing Wei,Hao Fan,Jing Zhang
In this paper, we first synthesized a novel tetraferrocene signal marker. Then a reagentless triplex DNA junctions-based electrochemical DNA sensor using signal amplification strategy of CHA and tetraferrocene was constructed. Tetraferrocene labeled hairpin probes (H1 and H2) were integrated together to form triplex DNA junctions through connecting chain (C1). It enabled catalytic hairpin assembly (CHA) reaction to be initiated without the addition of exogenous probe during the detection process, and the conformational change of the target caused the change of electrochemical signal. Combined with tetraferrocene and CHA strategies, the sensor achieved double signal amplification with a detection limit as low as 0.085pM. In addition, the sensor was successfully applied to mRNA detection in cells with high sensitivity and reagentless operation, providing possibilities for in vivo testing.
研究领域:
生物化学
·
分子生物学
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分析化学
·
生物技术
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DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2018
·
IF 5.667
Qian Kuang,Chunli Li,Zhiwei Qiu,Guifen Jie,Shuyan Niu,Tingyu Huang
In this work, a target-triggered three-way junction (3-WJ) structure was combined with enzyme-powered cascade amplification strategy for ultrasensitive electrochemiluminescence (ECL) detection of microRNA via free-running DNA walker. In the presence of target microRNA-21 (miRNA-21), a three-way junction structure was formed to trigger enzyme-aided multiple DNA amplification, and thus exponentially amplified triggers were generated. Then, the trigger DNA could hybridize with the blocking DNA on the electrode to release walker DNA, which specifically combined with the quantum dots (QDs)-DNA probe, and formed a recognition site for Nt.BbvCI. The movement of DNA walker is powered by the nicking endonuclease that cleaves specific QDs-DNA probes on the track. Thus one walker DNA can automatically cleave multiple QDs probes on the electrode during the movement, resulting in significantly amplified changes of ECL signal, which could be used for the highly sensitive detection of target miRNA-21. Under the optimal conditions, the ECL signal change is linear with the concentration of miRNA-21 in the range from 10 −14 M to 10 -7 M with a detection limit of 1.5 fM. This newly established strategy could achieve rapid, isothermal, and homogeneous signal amplification for specific nucleic acids in complicated biomatrix, which hold great potential for application in early clinical diagnosis.
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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产品关联
DOI
高影响因子
Journal of Physical Chemistry Letters
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2022
·
IF 6.888
Xianjing Zhang,Zhi Zhu,Wenpeng Liu,Feng Gao,Jun Guo,Bo Song,Luke P. Lee,Feng Zhang
Metal ions play vital roles in the electron transfer between biological molecules in humans, animals, and plants. However, the electron transfer between metal ions and nucleic acids and its impact on DNA–ion binding during DNA replication has been ignored. Here, we present a long-range quantum biological electron transfer (QBET) between DNA bases and metal ions and its selective function of DNA–ion binding in DNA replication. We discover biophysical DNA–ion binding and create biological filters that allow selective DNA replication by dual modulators of the valence and concentration of metal ions. QBET-based DNA replication filters provide powerful tools for ultrasensitive polymerase chain reaction (PCR) to selectively amplify target sequences with a discrete concentration window of metal ions; for example, Au3+ exhibits a concentration window that is approximately 3 orders of magnitude lower than that of Na+. DNA–ion filters provide new perspectives into metal ion-mediated QBET in DNA replication and hold great potential in life sciences and medical applications.
研究领域:
生物物理学
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分子生物学
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量子生物学
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生物无机化学
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遗传学
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DOI
高影响因子
ANALYTICAL CHEMISTRY
·
2025
·
IF 6.7
Jialin Zhang,Biyao Mao,Shuoshuo Cheng,Kangqiang Lu,Herui Wen,Jiali Ren
Assessing the activity of DNA methyltransferases (MTases) and screening for methyltransferase inhibitors not only allow for a deep exploration of the role of methylation regulation in disease initiation and progression but also provide an important experimental and clinical basis for the diagnosis and treatment of diseases. Herein, a new COFs functionalized electrochemical biosensor has been developed to detect DNA adenine methylation (Dam) MTase activity with high sensitivity and rapidity by taking advantage of the DNA walker and rolled circular strand displacement amplification (RC-SDA) reaction. Specifically, hairpin probe H1 was methylated by Dam MTase, followed by methylation site-specific cleavage of DpnI enzyme to generate the S5 probe. The padlock probes with two nucleic acid endonuclease sites were introduced to trigger the RC-SDA reaction under the action of the primer, releasing a large number of single-stranded S6 probes. The released probe worked synergistically with the Exo III enzyme to trigger DNA walking, which exposed the binding sites to enable the Au-COF-MB signal probes to bind effectively to the electrode surface, and the electrochemical signals were thus generated. The result showed that the designed electrochemical biosensor demonstrated excellent sensitivity and specificity in detecting the activity of Dam MTase, with the detection limit (LOD) of 6.85 × 10–5 U/mL. This method provides support for the development of new treatment strategies for methylation related diseases.
研究领域:
分子生物学
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分析化学
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医学诊断
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2015
·
IF 5.636
Nannan Liu,Zekun Yang,Xiaoding Lou,Benmei Wei,Juntao Zhang,Pengcheng Gao,Ruizuo Hou,Fan Xia
Almost all of the important functions of DNA are realized by proteins which interact with specific DNA, which actually happens in a limited space. However, most of the studies about the protein–DNA binding are in an unconfined space. Here, we propose a new method, nanopore-based DNA-probe sequence-evolution (NDPSE), which includes up to 6 different DNA-probe systems successively designed in a nanoscale confined space which unveil the more realistic characteristics of protein–DNA binding phenomena. There are several features; for example, first, the edge-hindrance and core-hindrance contribute differently for the binding events, and second, there is an equilibrium between protein–DNA binding and DNA–DNA hybridization.
研究领域:
分子生物学
·
生物信息学
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生物物理学
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产品关联
DOI
PubMed
高影响因子
BIOCHEMISTRY
·
2018
·
IF 2.997
Qianwei Zhao,Wen Yang,Tong Qin,Zhen Huang
RNA is a key player in the cellular central dogma, including RNA transcription and protein synthesis. However, it is unknown whether RNA can directly interfere with DNA synthesis. Recently, we have found in vitro that while binding to DNA polymerase nonspecifically, RNA can transform DNA polymerase to display a moonlighting activity, dNTP phosphatase, in turn interfering with DNA synthesis. This phosphatase activity removes the γ-phosphate from dNTPs (generating dNDPs) and subsequently removes the β-phosphate from the formed dNDPs (generating dNMPs), confirmed by the noncleavable α,β-CH2-dGTP and β,γ-CH2-dGTP analogues. We also found that dGTP is the best substrate for the phosphatase, and the dNTP phosphatase activity is sensitive to the reaction medium. In addition, we have revealed that RNA can tune the activity of closely related proteins and give rise to new catalytic functions with subtle differences. Moreover, we have demonstrated in vitro that at the lower dNTP level, this phosphatase can directly inhibit DNA synthesis by dNTP depletion, though the phosphatase activity is 690-fold slower than the polymerase activity. Our observation in vitro suggests a plausible strategy for RNA to directly interfere with DNA polymerase and DNA synthesis in vivo.
研究领域:
分子生物学
·
生物化学
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DOI
PubMed
高影响因子
TALANTA
·
2017
·
IF 4.162
Jing Zhu,Qijie Hao,Yi Liu,Zhaohui Guo,Buayxigul Rustam,Wei Jiang
The detection of uracil-DNA glycosylase (UDG) activity is pivotal for its biochemical studies and the development of drugs for UDG-related diseases. Here, we explored an integrated DNA structure switch for high sensitive detection of UDG activity. The DNA structure switch containing two branched hairpins was employed to recognize UDG enzyme and generate fluorescent signal. Under the action of UDG, one branched hairpin was impelled folding into a close conformation after the excision of the single uracil . This reconfigured hairpin could immediately initiate the polymerization/nicking amplification reaction of another branched hairpin accompanying with the release of numerous G-quadruplexes (G4s). In the absence of UDG, the DNA structure switch kept its original configuration, and thus the subsequent polymerization/nicking reaction was inhibited, resulting in the release of few G4 strands. In this work, Thioflavin T was used as signal reporter to target G4s. By integrating the DNA structure switch, the quick response and high sensitivity for UDG determination was achieved and a low detection limit of 0.0001 U/mL was obtained, which was superior to the most fluorescent methods for UDG assay. The repeatability of the as-proposed strategy was demonstrated under the concentration of 0.02 U/mL and 0.002 U/mL, the relative standard deviation obtained from 5 successive samples were 1.7% and 2.8%, respectively. The integrated DNA structure switch strategy proposed here has the potential application for the study of mechanism and function of UDG enzyme and the screening the inhibitors as potential drugs and biochemical tools.
研究领域:
生物化学
·
分子生物学
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分析化学
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药物开发
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产品关联
DOI
PubMed
高影响因子
Biomaterials Science
·
2017
·Vol. 5
·期 7
·1279-1292
·
IF 4.21
Yawen Liu,Zhaozhu Zheng,He Gong,Meng Liu,Shaozhe Guo,Gang Li,Xiaoqin Wang,David L. Kaplan
The structure of DNA is susceptible to alterations at high temperature and on changing pH, irradiation and exposure to DNase. Options to protect and preserve DNA during storage are important for applications in genetic diagnosis, identity authentication, drug development and bioresearch. In the present study, the stability of total DNA purified from human dermal fibroblast cells, as well as that of plasmid DNA, was studied in silk protein materials. The DNA/silk mixtures were stabilized on filter paper (silk/DNA + filter) or filter paper pre-coated with silk and treated with methanol (silk/DNA + PT-filter) as a route to practical utility. After air-drying and water extraction, 50–70% of the DNA and silk could be retrieved and showed a single band on electrophoretic gels. 6% silk/DNA + PT-filter samples provided improved stability in comparison with 3% silk/DNA + filter samples and DNA + filter samples for DNA preservation, with ∼40% of the band intensity remaining at 37 °C after 40 days and ∼10% after exposure to UV light for 10 hours. Quantitative analysis using the PicoGreen assay confirmed the results. The use of Tris/borate/EDTA (TBE) buffer enhanced the preservation and/or extraction of the DNA. The DNA extracted after storage maintained integrity and function based on serving as a functional template for PCR amplification of the gene for zinc finger protein 750 (ZNF750) and for transgene expression of red fluorescence protein (dsRed) in HEK293 cells. The high molecular weight and high content of a crystalline beta-sheet structure formed on the coated surfaces likely accounted for the preservation effects observed for the silk/DNA + PT-filter samples. Although similar preservation effects were also obtained for lyophilized silk/DNA samples, the rapid and simple processing available with the silk–DNA–filter membrane system makes it appealing for future applications.
研究领域:
遗传学
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生物技术
·
材料科学
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2022
·
IF 9.221
Hui Zhou,Hui Shen,Baoting Dou,Qiumei Feng,Xiguang Han,Po Wang
Methylated DNA is a promising epigenetic biomarker for diseases diagnosis, therapy monitoring, and prognosis. Here, we developed a sensitive ratiometric electrochemical biosensor for the detection of methylated DNA based on the design of multistep DNA amplification circuits. The methylated target DNA firstly initiates the DNAzyme-assisted amplification circuit and produces a number of single-stranded triggers that catalyze the subsequent double-step catalytic hairpin assembly circuits to release numerous four-way DNA junctions. The four-way DNA junctions displace the methylene blue (MB)-modified signal probes for intercalating doxorubicin (DOX) molecules on the sensing interface, resulting in a significantly increased value of i DOX / i MB to achieve sensitive monitoring of methylated DNA. Taking advantage of the significant signal amplification of multistep DNA amplification circuits, the proposed method exhibited high sensitivity for target analysis with a detection limit of 4 aM and a dynamic linear range of 10 aM to 20 pM. Moreover, the proof-of-concept application of the sensing platform was investigated, suggesting that the system is promising for potential applications in early cancer diagnosis and the fundamental research of epigenetics.
研究领域:
分子生物学
·
医学检测
·
纳米生物技术
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产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）