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  1 / 806 MEDLINE  
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[PMID]:27706624
[Au] Autor:Barbosa AM; de Souza SR; Frare AB; Costa E Silva RC; da Costa IR; Freitas E Silva KS; Ribeiro Júnior CL; Bordin BM; Moura KK
[Ad] Endereço:Departamento de Biologia, Núcleo de Pesquisas Replicon, Pontifícia Universidade Católica de Goiás, Goiânia, GO, Brasil andreiamarcelino_@hotmail.com.
[Ti] Título:Association of CYP1A1 (cytochrome P450) MspI polymorphism in women with endometriosis.
[So] Source:Genet Mol Res;15(3), 2016 Aug 26.
[Is] ISSN:1676-5680
[Cp] País de publicação:Brazil
[La] Idioma:eng
[Ab] Resumo:Endometriosis is a disease that affects 10 to 15% of the women of reproductive age. It is characterized by the presence of endometrial-like tissues outside of the uterus. Some definitions claim that the functional ectopic tissue is sensitive to the action of hormones. Severity of endometriosis is defined according to a system proposed by the American Society for Reproductive Medicine, which is based on laparoscopic findings. A large number of genetic polymorphisms has been reported for CYP1A1, the gene that is responsible for enzymes involved in stage I detoxification of xenobiotics; this gene is located at 15q22-24, and encodes an isoenzyme that catalyzes the oxidation of polycyclic aromatic hydrocarbons present in phenolic compounds and epoxides. The aim of this study was to analyze the frequency of the MspI polymorphism and its relation to endometriosis. We obtained peripheral blood samples from 52 women with endometriosis (confirmed by laparoscopy) as well as 42 women without endometriosis (control group). In the case group, the women were between 25 and 35 years of age; the age range was between 25 and 57 years old in the control group. Molecular analysis was performed by polymerase chain reaction. We found a significant association (P = 0.039) between the polymorphic allele m1 and endometriosis (32.70%). In conclusion, this study showed that the m1 polymorphism is associated with endometriosis, and that W1/m1 and m1/m1 polymorphisms are more frequently observed in patients with infertility and severe endometriosis.
[Mh] Termos MeSH primário: Citocromo P-450 CYP1A1/genética
Desoxirribonuclease HpaII/química
Endometriose/genética
Infertilidade Feminina/genética
Polimorfismo de Fragmento de Restrição
[Mh] Termos MeSH secundário: Adulto
Alelos
Estudos de Casos e Controles
Endometriose/complicações
Endometriose/diagnóstico
Endometriose/patologia
Feminino
Expressão Gênica
Frequência do Gene
Genótipo
Seres Humanos
Infertilidade Feminina/complicações
Infertilidade Feminina/diagnóstico
Infertilidade Feminina/patologia
Leucócitos Mononucleares/metabolismo
Leucócitos Mononucleares/patologia
Meia-Idade
Índice de Gravidade de Doença
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
EC 1.14.14.1 (CYP1A1 protein, human); EC 1.14.14.1 (Cytochrome P-450 CYP1A1); EC 3.1.21.- (Deoxyribonuclease HpaII)
[Em] Mês de entrada:1702
[Cu] Atualização por classe:170224
[Lr] Data última revisão:
170224
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161006
[St] Status:MEDLINE
[do] DOI:10.4238/gmr.15038389


  2 / 806 MEDLINE  
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[PMID]:26727463
[Au] Autor:Liu G; Weston CQ; Pham LK; Waltz S; Barnes H; King P; Sphar D; Yamamoto RT; Forsyth RA
[Ad] Endereço:FLIR Systems, Inc., La Jolla, California, 92037, United States of America.
[Ti] Título:Epigenetic Segregation of Microbial Genomes from Complex Samples Using Restriction Endonucleases HpaII and McrB.
[So] Source:PLoS One;11(1):e0146064, 2016.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:We describe continuing work to develop restriction endonucleases as tools to enrich targeted genomes of interest from diverse populations. Two approaches were developed in parallel to segregate genomic DNA based on cytosine methylation. First, the methyl-sensitive endonuclease HpaII was used to bind non-CG methylated DNA. Second, a truncated fragment of McrB was used to bind CpG methylated DNA. Enrichment levels of microbial genomes can exceed 100-fold with HpaII allowing improved genomic detection and coverage of otherwise trace microbial genomes from sputum. Additionally, we observe interesting enrichment results that correlate with the methylation states not only of bacteria, but of fungi, viruses, a protist and plants. The methods presented here offer promise for testing biological samples for pathogens and global analysis of population methylomes.
[Mh] Termos MeSH primário: 5-Metilcitosina/análise
Enzimas de Restrição do DNA
DNA Bacteriano/isolamento & purificação
DNA Fúngico/isolamento & purificação
DNA de Plantas/isolamento & purificação
DNA de Protozoário/isolamento & purificação
DNA Viral/isolamento & purificação
Desoxirribonuclease HpaII
Proteínas de Escherichia coli
Genética Microbiana/métodos
Genômica/métodos
Metagenoma
[Mh] Termos MeSH secundário: Ilhas de CpG/genética
Metilação de DNA
Enzimas de Restrição do DNA/isolamento & purificação
Enzimas de Restrição do DNA/metabolismo
DNA Bacteriano/genética
DNA Fúngico/genética
DNA de Plantas/genética
DNA de Protozoário/genética
DNA Viral/genética
Desoxirribonuclease HpaII/isolamento & purificação
Desoxirribonuclease HpaII/metabolismo
Proteínas de Escherichia coli/isolamento & purificação
Proteínas de Escherichia coli/metabolismo
Biblioteca Gênica
Seres Humanos
Microbiota/genética
Análise de Sequência de DNA
Escarro/microbiologia
Especificidade por Substrato
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, U.S. GOV'T, NON-P.H.S.
[Nm] Nome de substância:
0 (DNA, Bacterial); 0 (DNA, Fungal); 0 (DNA, Plant); 0 (DNA, Protozoan); 0 (DNA, Viral); 0 (Escherichia coli Proteins); 6R795CQT4H (5-Methylcytosine); EC 3.1.21.- (DNA Restriction Enzymes); EC 3.1.21.- (Deoxyribonuclease HpaII); EC 3.1.21.- (mcrB protein, E coli)
[Em] Mês de entrada:1607
[Cu] Atualização por classe:170220
[Lr] Data última revisão:
170220
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160105
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0146064


  3 / 806 MEDLINE  
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[PMID]:26841491
[Au] Autor:Kulibaba RA; Yurko PS; Liashenkon YV
[Ti] Título:[MspI-POLYMORPHISM IN FOURTH INTRON OF THE GROWTH HORMONE GENE IN CHICKEN POPULATIONS OF DIFFERENT BREEDS. ANALYSIS OF THE CAUSES OF ADDITIONAL RESTRICTION PATTERN ORIGIN].
[So] Source:Tsitol Genet;49(6):30-7, 2015 Nov-Dec.
[Is] ISSN:0564-3783
[Cp] País de publicação:Ukraine
[La] Idioma:rus
[Ab] Resumo:The MspI-polymorphism in the fourth intron of the growth hormone gene in populations of White Plymouth Rock, Poltava Clay, Rhode Island Red and Borkovskaya Barvistaya chicken breeds was studied. It is shown that in all examined chicken populations the growth hormone gene is polymorphic. It was found that the presence of "additional" phenotype (restriction pattern) is not associated with duplication of the growth hormone gene. The possibility of formation of heteroduplex DNA of two different types in the course of amplification of heterozygous samples B/C, containing the site 'CCGG', which leads to formation the additional DNA fragment which do not contain the site 'CCGG', was described. The nucleotide sequences of alleles A, B, C and "additional" fragment is described. Frequencies of alleles A, B and C in chicken population of White Plymouth Rock breed were 0.56; 0.16 and 0.28; Poltava Clay--0.10; 0.07 and 0.83; Rhode Island Red--0.27; 0.31 and 0.42; Borkovskaya Barvistaya--0.75; 0.08 and 0.17 respectively.
[Mh] Termos MeSH primário: Galinhas/genética
Desoxirribonuclease HpaII/metabolismo
Hormônio do Crescimento/genética
Íntrons
Polimorfismo de Fragmento de Restrição
[Mh] Termos MeSH secundário: Animais
Sequência de Bases
Cruzamento
Eletroforese em Gel de Ágar
Frequência do Gene
Dados de Sequência Molecular
Ácidos Nucleicos Heteroduplexes/genética
Reação em Cadeia da Polimerase
[Pt] Tipo de publicação:ENGLISH ABSTRACT; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Nucleic Acid Heteroduplexes); 9002-72-6 (Growth Hormone); EC 3.1.21.- (Deoxyribonuclease HpaII)
[Em] Mês de entrada:1603
[Cu] Atualização por classe:160204
[Lr] Data última revisão:
160204
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160205
[St] Status:MEDLINE


  4 / 806 MEDLINE  
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[PMID]:26267039
[Au] Autor:Liu Y; Wei M; Zhang L; Wei W; Zhang Y; Liu S
[Ad] Endereço:Jiangsu Province Hi-Tech Key Laboratory for Bio-medical Research, School of Chemistry and Chemical Engineering, Southeast University, Nanjing, 211189, China. wei_wei98@163.com.
[Ti] Título:Evaluation of DNA methyltransferase activity and inhibition via chiroplasmonic assemblies of gold nanoparticles.
[So] Source:Chem Commun (Camb);51(76):14350-3, 2015 Oct 01.
[Is] ISSN:1364-548X
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Circular dichroism spectroscopy has been explored for detection of methyltransferase activity and inhibition based on DNA-induced chiroplasmonic assemblies of gold nanoparticles and endonuclease HpaII. Good accuracy, precision and sensitivity are obtained in complex matrices such as human serum samples, which is significant for clinical diagnosis and drug development.
[Mh] Termos MeSH primário: Dicroísmo Circular/métodos
Desoxirribonuclease HpaII/sangue
Desoxirribonuclease HpaII/metabolismo
Ouro/química
Nanopartículas Metálicas/química
[Mh] Termos MeSH secundário: DNA/metabolismo
DNA-Citosina Metilases/antagonistas & inibidores
DNA-Citosina Metilases/sangue
DNA-Citosina Metilases/metabolismo
Desoxirribonuclease HpaII/antagonistas & inibidores
Dimerização
Avaliação Pré-Clínica de Medicamentos/métodos
Ensaios Enzimáticos/métodos
Inibidores Enzimáticos/farmacologia
Seres Humanos
Nanopartículas Metálicas/ultraestrutura
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Enzyme Inhibitors); 7440-57-5 (Gold); 9007-49-2 (DNA); EC 2.1.1.- (DNA modification methylase SssI); EC 2.1.1.- (DNA-Cytosine Methylases); EC 3.1.21.- (Deoxyribonuclease HpaII)
[Em] Mês de entrada:1606
[Cu] Atualização por classe:150910
[Lr] Data última revisão:
150910
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:150813
[St] Status:MEDLINE
[do] DOI:10.1039/c5cc05375g


  5 / 806 MEDLINE  
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[PMID]:26070170
[Au] Autor:Zhang L; Wei M; Gao C; Wei W; Zhang Y; Liu S
[Ad] Endereço:Jiangsu Province Hi-Tech Key Laboratory for Bio-medical Research, School of Chemistry and Chemical Engineering, Southeast University, Nanjing 211189, PR China; Analytical & Testing Center of Nanjing Normal University, Nanjing 210023, PR China.
[Ti] Título:Label-free electrochemical detection of methyltransferase activity and inhibitor screening based on endonuclease HpaII and the deposition of polyaniline.
[So] Source:Biosens Bioelectron;73:188-94, 2015 Nov 15.
[Is] ISSN:1873-4235
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Detection of DNA methylation and methyltransferase (MTase) activity are important in determining human cancer because aberrant methylation was linked to cancer initiation and progression. In this work, we proposed an electrochemical method for sensitive detection of DNA methylation and MTase activity based on methylation sensitive restriction endonuclease HpaII and the deposition of polyaniline (PANI) catalyzed by HRP-mimicking DNAzyme. In the presence of methylated DNA, HRP-mimicking DNAzyme catalyzed the polymerization of aniline on the dsDNA template, producing huge DPV current. In the presence of non-methylated DNA, dsDNA are cleaved and digested by HpaII and exonuclease III, as a result, no PANI are deposited. This method can be used to determine DNA methylation at the site of CpG. It exhibits a wide linear response toward M.SssI MTase activity in the range of 0.5-0.6 U mL(-1) with the detection limit of 0.12 U mL(-1). G-rich DNA forms HRP mimicking DNAzyme, which avoids complex labeling procedures and is robust. The method is simple, reliable, sensitive and specific, which has been successfully applied in human serum samples and been used to screen the inhibitors. Thus, the proposed method may be a potential and powerful tool for clinical diagnosis and drug development in the future.
[Mh] Termos MeSH primário: Técnicas Biossensoriais/métodos
Metilação de DNA
DNA-Citosina Metilases/análise
Técnicas Eletroquímicas/métodos
[Mh] Termos MeSH secundário: Compostos de Anilina
Sondas de DNA
DNA Catalítico
DNA-Citosina Metilases/antagonistas & inibidores
DNA-Citosina Metilases/sangue
Desoxirribonuclease HpaII
Avaliação Pré-Clínica de Medicamentos
Inibidores Enzimáticos/farmacologia
Seres Humanos
Limite de Detecção
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Aniline Compounds); 0 (DNA Probes); 0 (DNA, Catalytic); 0 (Enzyme Inhibitors); 0 (polyaniline); EC 2.1.1.- (DNA modification methylase SssI); EC 2.1.1.- (DNA-Cytosine Methylases); EC 3.1.21.- (Deoxyribonuclease HpaII)
[Em] Mês de entrada:1604
[Cu] Atualização por classe:150720
[Lr] Data última revisão:
150720
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:150613
[St] Status:MEDLINE


  6 / 806 MEDLINE  
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[PMID]:25886559
[Au] Autor:Zheng H; Zhao Y
[Ad] Endereço:Nursing Department, Tai'an Tumor Hospital, Tai'an City, No. 262 Taidong Road, Shandong Province, 271000, China. Huizheng2000@126.com.
[Ti] Título:Association of CYP1A1 MspI polymorphism in the esophageal cancer risk: a meta-analysis in the Chinese population.
[So] Source:Eur J Med Res;20:46, 2015 Mar 30.
[Is] ISSN:2047-783X
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Although many epidemiologic studies have investigated the CYP1A1 MspI gene polymorphisms and their associations with esophageal cancer (EC), definite conclusions cannot be drawn. To clarify the effects of CYP1A1 MspI polymorphisms on the risk of EC, a meta-analysis was performed in Chinese population. METHODS: Related studies were identified from PubMed, Springer Link, Ovid, Chinese Wanfang Data Knowledge Service Platform, Chinese National Knowledge Infrastructure (CNKI), and Chinese Biology Medicine (CBM) till October 2014. Pooled ORs and 95% CIs were used to assess the strength of the associations. RESULTS: A total of 13 studies including 1,519 EC cases and 1,962 controls were involved in this meta-analysis. Overall, significant association was found between CYP1A1 MspI polymorphism and EC risk when all studies in the Chinese population pooled into this meta-analysis (C vs. T: OR = 1.25, 95% CI = 1.04 to 1.51; CC + CT vs. TT: OR = 1.35, 95% CI = 1.06 to 1.72; CC vs. TT + CT: OR = 1.35, 95% CI = 1.03 to 1.76). When we performed stratified analyses by geographical locations, histopathology type, and source of control, significantly increased risks were found in North China (C vs. T: OR = 1.38, 95% CI = 1.12 to 1.70; CC vs. TT: OR = 1.72, 95% CI = 1.16 to 2.56; CC + CT vs. TT: OR = 1.52, 95% CI = 1.14 to 2.02; CC vs. TT + CT: OR = 1.55, 95% CI = 1.17 to 2.06), in the population-based studies (C vs. T: OR = 1.22, 95% CI = 1.05 to 1.42; CC vs. TT: OR = 1.38, 95% CI = 1.02 to 1.88; CC + CT vs. TT: OR = 1.36, 95% CI = 1.10 to 1.69; CC vs. TT + CT: OR = 1.43, 95% CI = 1.13 to 1.81) and ESCC (C vs. T: OR = 1.17, 95% CI = 1.04 to 1.32; CC + CT vs. TT: OR = 1.28, 95% CI = 1.08 to 1.52). CONCLUSIONS: This meta-analysis provides the evidence that CYP1A1 MspI polymorphism may contribute to the EC development in the Chinese population.
[Mh] Termos MeSH primário: Citocromo P-450 CYP1A1/genética
Desoxirribonuclease HpaII/genética
Neoplasias Esofágicas/genética
Predisposição Genética para Doença
Polimorfismo Genético
[Mh] Termos MeSH secundário: China
Seres Humanos
[Pt] Tipo de publicação:JOURNAL ARTICLE; META-ANALYSIS
[Nm] Nome de substância:
EC 1.14.14.1 (CYP1A1 protein, human); EC 1.14.14.1 (Cytochrome P-450 CYP1A1); EC 3.1.21.- (Deoxyribonuclease HpaII)
[Em] Mês de entrada:1602
[Cu] Atualização por classe:170220
[Lr] Data última revisão:
170220
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:150418
[St] Status:MEDLINE
[do] DOI:10.1186/s40001-015-0135-3


  7 / 806 MEDLINE  
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[PMID]:25421665
[Au] Autor:Hahn MA; Li AX; Wu X; Pfeifer GP
[Ad] Endereço:Department of Cancer Biology, Beckman Research Institute, City of Hope, Duarte, CA, USA, mhahn@coh.org.
[Ti] Título:Single base resolution analysis of 5-methylcytosine and 5-hydroxymethylcytosine by RRBS and TAB-RRBS.
[So] Source:Methods Mol Biol;1238:273-87, 2015.
[Is] ISSN:1940-6029
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Sodium bisulfite-assisted deamination of cytosine forms the basis for conducting single base resolution analysis of 5-methylcytosine in DNA. The TET family of proteins represents a group of enzymes that can oxidize 5-methylcytosine to 5-hydroxymethylcytosine. A modification of the bisulfite-based DNA methylation mapping technique employs TET1-mediated oxidation of 5-methylcytosine (TET-assisted bisulfite sequencing) for single base analysis of 5-hydroxymethylcytosine. Whole genome analysis of cytosine modifications with bisulfite sequencing techniques still is challenging and expensive. Reduced representation bisulfite sequencing (RRBS) has been used to limit the complexity of the analysis to mostly CpG-rich genomic fragments flanked by restriction enzyme cleavage sites, for example MspI (5'CCGG). In this chapter, we describe detailed methods used in our laboratory for analysis of 5-methylcytosine and 5-hydroxymethylcytosine combined (RRBS) and for specific analysis of 5-hydroxymethylcytosine (TAB-RRBS).
[Mh] Termos MeSH primário: 5-Metilcitosina/metabolismo
Citosina/análogos & derivados
Análise de Sequência de DNA/métodos
[Mh] Termos MeSH secundário: Citosina/metabolismo
Desoxirribonuclease HpaII/metabolismo
Glicosilação
Seres Humanos
Oxirredução
Sulfitos/farmacologia
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, N.I.H., EXTRAMURAL
[Nm] Nome de substância:
0 (Sulfites); 1123-95-1 (5-hydroxymethylcytosine); 6R795CQT4H (5-Methylcytosine); 8J337D1HZY (Cytosine); EC 3.1.21.- (Deoxyribonuclease HpaII); TZX5469Z6I (sodium bisulfite)
[Em] Mês de entrada:1507
[Cu] Atualização por classe:161019
[Lr] Data última revisão:
161019
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:141126
[St] Status:MEDLINE
[do] DOI:10.1007/978-1-4939-1804-1_14


  8 / 806 MEDLINE  
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[PMID]:25408952
[Au] Autor:Zhang C; Lou J; Tu W; Bao J; Dai Z
[Ad] Endereço:Jiangsu Key Laboratory of Biofunctional Materials, College of Chemistry and Materials Science, Nanjing Normal University, Nanjing, 210023, P. R. China. daizhihuii@njnu.edu.cn.
[Ti] Título:Ultrasensitive electrochemical biosensing for DNA using quantum dots combined with restriction endonuclease.
[So] Source:Analyst;140(2):506-11, 2015 Jan 21.
[Is] ISSN:1364-5528
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:A universal and sensitive electrochemical biosensing platform for the detection and identification of DNA using CdSe quantum dots (CdSe QDs) as signal markers was designed. The detection mechanism was based on the specific recognition of MspI endonuclease combined with the signal amplification of gold nanoparticles (AuNPs). MspI endonuclease could recognize its specific sequence in the double-strand DNA (dsDNA) and cleave the dsDNA fragments linked with CdSe QDs from the electrode. The remaining attached CdSe QDs can be easily read out by square-wave voltammetry using an electrodeposited bismuth (Bi) film-modified glass carbon electrode. The concentrations of target DNA could be simultaneously detected by the signal of metal markers. Using mycobacterium tuberculosis (Mtb) DNA as a model, under the optimal conditions, the proposed biosensor could detect Mtb DNA down to 8.7 × 10(-15) M with a linear range of 5 orders of magnitude (from 1.0 × 10(-14) to 1.0 × 10(-9) M) and discriminate mismatched DNA with high selectivity. This strategy presented a universal and convenient biosensing platform for DNA assay, and its satisfactory performances make it a potential candidate for the early diagnosis of gene-related diseases.
[Mh] Termos MeSH primário: Técnicas Biossensoriais/métodos
DNA Bacteriano/análise
Desoxirribonuclease HpaII/metabolismo
Pontos Quânticos/química
Tuberculose Pulmonar/diagnóstico
[Mh] Termos MeSH secundário: Compostos de Cádmio/química
DNA Bacteriano/genética
Técnicas Eletroquímicas/métodos
Ouro/química
Nanopartículas Metálicas/química
Mycobacterium tuberculosis/genética
Compostos de Selênio/química
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Cadmium Compounds); 0 (DNA, Bacterial); 0 (Selenium Compounds); 7440-57-5 (Gold); A7F646JC5C (cadmium selenide); EC 3.1.21.- (Deoxyribonuclease HpaII)
[Em] Mês de entrada:1510
[Cu] Atualização por classe:141216
[Lr] Data última revisão:
141216
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:141120
[St] Status:MEDLINE
[do] DOI:10.1039/c4an01284d


  9 / 806 MEDLINE  
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[PMID]:25281112
[Au] Autor:Wei W; Gao C; Xiong Y; Zhang Y; Liu S; Pu Y
[Ad] Endereço:Key Laboratory of Environmental Medicine Engineering, Ministry of Education, School of Chemistry and Chemical Engineering, Southeast University, Jiangning District, Nanjing 211189, PR China.
[Ti] Título:A fluorescence method for detection of DNA and DNA methylation based on graphene oxide and restriction endonuclease HpaII.
[So] Source:Talanta;131:342-7, 2015 Jan.
[Is] ISSN:1873-3573
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:DNA methylation plays an important role in many biological events and is associated with various diseases. Most traditional methods for detection of DNA methylation are based on the complex and expensive bisulfite method. In this paper, we report a novel fluorescence method to detect DNA and DNA methylation based on graphene oxide (GO) and restriction endonuclease HpaII. The skillfully designed probe DNA labeled with 5-carboxyfluorescein (FAM) and optimized GO concentration keep the probe/target DNA still adsorbed on the GO. After the cleavage action of HpaII the labeled FAM is released from the GO surface and its fluorescence recovers, which could be used to detect DNA in the linear range of 50 pM-50 nM with a detection limit of 43 pM. DNA methylation induced by transmethylase (Mtase) or other chemical reagents prevents HpaII from recognizing and cleaving the specific site; as a result, fluorescence cannot recover. The fluorescence recovery efficiency is closely related to the DNA methylation level, which can be used to detect DNA methylation by comparing it with the fluorescence in the presence of intact target DNA. The method for detection of DNA and DNA methylation is simple, reliable and accurate.
[Mh] Termos MeSH primário: Metilação de DNA
DNA/análise
Desoxirribonuclease HpaII/metabolismo
Corantes Fluorescentes/química
Grafite/química
[Mh] Termos MeSH secundário: Sondas de DNA/química
Fluoresceínas/química
Fluorescência
Limite de Detecção
Espectrometria de Fluorescência
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (DNA Probes); 0 (Fluoresceins); 0 (Fluorescent Dyes); 76823-03-5 (4-carboxyfluorescein); 7782-42-5 (Graphite); 9007-49-2 (DNA); EC 3.1.21.- (Deoxyribonuclease HpaII)
[Em] Mês de entrada:1506
[Cu] Atualização por classe:141004
[Lr] Data última revisão:
141004
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:141005
[St] Status:MEDLINE


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[PMID]:25343162
[Au] Autor:Gao C; Li H; Liu Y; Wei W; Zhang Y; Liu S
[Ad] Endereço:Key Laboratory of Environmental Medicine Engineering, Ministry of Education, School of Chemistry and Chemical Engineering, Southeast University, Jiangning District, Nanjing, 211189, P.R. China. wei_wei98@163.com.
[Ti] Título:Label-free fluorescence detection of DNA methylation and methyltransferase activity based on restriction endonuclease HpaII and exonuclease III.
[So] Source:Analyst;139(24):6387-92, 2014 Dec 21.
[Is] ISSN:1364-5528
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Strategies to detect the methylation of site specific DNA and assay of M.SssI methyltransferase (M.SssI MTase) activity are important in determining human cancers due to aberrant methylation linked to cancer initiation and progression. Herein, we report a label-free fluorescence detection method for DNA methylation and MTase activity based on restriction endonuclease HpaII and exonuclease III (Exo III). A label-free probe DNA was designed, which hybridized with target DNA (one 32-mer DNA from the exon 8 promoter region of the Homo sapiens p53 gene) to form double stranded DNA (dsDNA). Upon the cleavage action of HpaII and degradation reaction of Exo III, dsDNA changed to single stranded DNA (ssDNA) and the fluorescence intensity of thiazole orange (TO) is weak. After the resulting dsDNA was methylated by M.SssI MTase, the action of HpaII and Exo III was prevented, then TO intercalates into the dsDNA and emits strong fluorescence. This method can determine DNA methylation at the site of CpG and distinguish a one-base mismatched target sequence. The fluorescence intensity has a linear relationship with M.SssI MTase activities in the range of 1-10 U mL(-1) with a detection limit of 0.16 U mL(-1) in terms of 3 times deviation of the blank sample. The methylation of DNA by a hydroxyl radical triggered by DMSO and CH3CHO was also measured. These results show that the proposed method can specifically and selectively detect DNA methylation and M.SssI MTase activity. Human serum has no obvious effects on the assay performance, indicating that the method has great potential for further application in complex samples.
[Mh] Termos MeSH primário: Metilação de DNA
DNA-Citosina Metilases/metabolismo
DNA/genética
Desoxirribonuclease HpaII/metabolismo
Exodesoxirribonucleases/metabolismo
Genes p53
[Mh] Termos MeSH secundário: DNA/sangue
DNA/química
DNA/metabolismo
Ensaios Enzimáticos/métodos
Fluorescência
Seres Humanos
Regiões Promotoras Genéticas
Espectrometria de Fluorescência/métodos
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
9007-49-2 (DNA); EC 2.1.1.- (DNA modification methylase SssI); EC 2.1.1.- (DNA-Cytosine Methylases); EC 3.1.- (Exodeoxyribonucleases); EC 3.1.11.2 (exodeoxyribonuclease III); EC 3.1.21.- (Deoxyribonuclease HpaII)
[Em] Mês de entrada:1508
[Cu] Atualização por classe:141110
[Lr] Data última revisão:
141110
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:141025
[St] Status:MEDLINE
[do] DOI:10.1039/c4an01359j



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