Rapid and selective DNA-based detection of melamine using α-hemolysin nanopores

Sheng, Yingying; You, Yi; Cao, Zhong; Liu, Lei; Wu, Hai‐Chen

doi:10.1039/c8an00580j

Cited by 47 publications

(19 citation statements)

References 65 publications

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“…Liu with coauthors [34] used single-wall carbon nanotubes to selectively detect modified 5-hydroxymethylcytosine in single-stranded DNA, which can be used to screen specific genomic DNA. The proposed strategy can be extended to many other useful applications in the field of chemistry and biology [35]. Despite the rapid development of the technologies described above, the exact determination of nucleotides and the nature of methylation remain an ongoing issue.…”

Section: Resultsmentioning

confidence: 99%

An alternative approach to study the enzymatic specificities of the CfrBI restriction–modification system

Zakharova

Beletskaya

Ibryashkina

et al. 2019

Heliyon

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Restriction–modification systems (RMS) are the main gene-engineering tools and a suitable model to study the molecular mechanisms of catalysis and DNA–protein interactions. Research into the catalytic properties of these enzymes, determination of hydrolysis and DNA-methylation sites remain topical. In our previous work we have cloned and sequenced the CfrBI restriction–modification system (strain Citrobacter freundii) , which recognizes the nucleotide sequence 5′-CCWWGG-3′. In this article we describe the cloning of the methyltransferase and restriction endonuclease genes (gene encoding CfrBI DNA methyltransferase ( cfrBIM) and gene encoding CfrBI restriction endonuclease ( cfrBIR) ) separately to obtain strains overproducing the enzymes of this system. His 6 -CfrBI, which had been purified to homogeneity, was used to establish the DNA-hydrolysis point in its recognition site. CfrBI was shown to cleave DNA after just the first 5′C within the recognition site and then to generate 4-nt 3′ cohesive ends (5′-C/CWWGG-3′). To map the site of methylation by M.CfrBI, we exploited the fact that the Cfr BI site partially overlaps with the recognition sites of the well-documented enzymes KpnI and ApaI. The M.CfrBI- induced hemimethylation of the internal C residue of the ApaI recognition sequence (GGGC N4m CC) was observed to block cleavage by ApaI. In contrast, KpnI was able to digest its M.CfrBI-hemimethylated site (GGTA N4m CC). KpnI was used to restrict a fragment of DNA harbouring the CfrBI and KpnI sites, in which the CfrBI site was methylated in vitro by His 6 -M.CfrBI using [ 3 H]-SAM. The subsequent separation of hydrolysis products by electrophoresis and the enumeration of incorporated [H3]-methyl groups in each of the fragments made it possible to determine that external cytosine undergoes modification in the recognition site.

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Section: Resultsmentioning

confidence: 99%

An alternative approach to study the enzymatic specificities of the CfrBI restriction–modification system

Zakharova

Beletskaya

Ibryashkina

et al. 2019

Heliyon

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“…The α-Hemolysin could detect melamine which is a common food additive as low as 10 pM with the aid of DNA probes. [59] Under de novo design by performing mutagenesis, label-free detection is achievable. In the case of monosaccharides, D-glucose and D-fructose exist in aqueous solution as an equilibrium mixture.…”

Section: Nanopore Beyond Dna Sequencingmentioning

confidence: 99%

Single-entity electrochemistry at confined sensing interfaces

et al. 2020

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Measurements at the single-entity level provide more precise diagnosis and understanding of basic biological and chemical processes. Recent advances in the chemical measurement provide a means for ultra-sensitive analysis. Confining the single analyte and electrons near the sensing interface can greatly enhance the sensitivity and selectivity. In this review, we summarize the recent progress in single-entity electrochemistry of single molecules, single particles, single cells and even brain analysis. The benefits of confining these entities to a compatible size sensing interface are exemplified. Finally, the opportunities and challenges of single entity electrochemistry are addressed.

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“…However, to the best of our knowledge, the preparation of quinolin-2-yl substituted ureas from readily available quinoline N -oxides has never been reported. In continuation of our research program in green and sustainable chemistry, − we have reported the ecofriendly preparation of N -heterocycles and their derivatives. − In this paper, we report an environmentally friendly method for the clean preparation of quinolin-2-yl substituted ureas from readily available quinoline N -oxides and carbodiimides in water (Scheme d).…”

Section: Introductionmentioning

confidence: 99%

Clean Preparation of Quinolin-2-yl Substituted Ureas in Water

Xie

Peng

Liu

et al. 2019

ACS Sustainable Chem. Eng.

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An environmentally-friendly method for the clean preparation of various quinolin-2-yl substituted ureas in water under mild and toxic reagent-, base-, and organic solvent-free conditions was established. In the large-scale synthesis, the products could be rapidly collected via simple filtration and washing with ethanol. The usage of readily available raw materials, 100% atom economy, high yield, and excellent regioselectivity enhanced the practicability of this protocol.

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Rapid and selective DNA-based detection of melamine using α-hemolysin nanopores

Abstract: We have developed a rapid and selective approach for the detection of melamine based on simple DNA probes and α-hemolysin nanopores. The limit of detection can be as low as 10 pM.

Cited by 47 publications

References 65 publications

An alternative approach to study the enzymatic specificities of the CfrBI restriction–modification system

An alternative approach to study the enzymatic specificities of the CfrBI restriction–modification system

Single-entity electrochemistry at confined sensing interfaces

Clean Preparation of Quinolin-2-yl Substituted Ureas in Water

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