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[PMID]:29335514
[Au] Autor:Kaiser M; Jug F; Julou T; Deshpande S; Pfohl T; Silander OK; Myers G; van Nimwegen E
[Ad] Endereço:Biozentrum, University of Basel and Swiss Institute of Bioinformatics, Klingelbergstrasse 50/70, 4056, Basel, Switzerland.
[Ti] Título:Monitoring single-cell gene regulation under dynamically controllable conditions with integrated microfluidics and software.
[So] Source:Nat Commun;9(1):212, 2018 01 15.
[Is] ISSN:2041-1723
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Much is still not understood about how gene regulatory interactions control cell fate decisions in single cells, in part due to the difficulty of directly observing gene regulatory processes in vivo. We introduce here a novel integrated setup consisting of a microfluidic chip and accompanying analysis software that enable long-term quantitative tracking of growth and gene expression in single cells. The dual-input Mother Machine (DIMM) chip enables controlled and continuous variation of external conditions, allowing direct observation of gene regulatory responses to changing conditions in single cells. The Mother Machine Analyzer (MoMA) software achieves unprecedented accuracy in segmenting and tracking cells, and streamlines high-throughput curation with a novel leveraged editing procedure. We demonstrate the power of the method by uncovering several novel features of an iconic gene regulatory program: the induction of Escherichia coli's lac operon in response to a switch from glucose to lactose.
[Mh] Termos MeSH primário: Regulação Bacteriana da Expressão Gênica
Técnicas Analíticas Microfluídicas/métodos
Análise de Célula Única/métodos
Software
[Mh] Termos MeSH secundário: Algoritmos
Rastreamento de Células/instrumentação
Rastreamento de Células/métodos
Escherichia coli/citologia
Escherichia coli/efeitos dos fármacos
Escherichia coli/genética
Glucose/farmacologia
Óperon Lac/genética
Lactose/farmacologia
Análise de Célula Única/instrumentação
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
IY9XDZ35W2 (Glucose); J2B2A4N98G (Lactose)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180305
[Lr] Data última revisão:
180305
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180117
[St] Status:MEDLINE
[do] DOI:10.1038/s41467-017-02505-0


  2 / 6777 MEDLINE  
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[PMID]:29323842
[Au] Autor:Kuznetsova TV; Smimova MS; Leonovich OA; Gordeichuk IV; Biriukova IK; Zylkova MV; Tyn'o YY; Belyakova AV; Shevelev AB
[Ti] Título:A new method of producing NS5A antigen of hepatitis C virus.
[So] Source:Vopr Virusol;62(1):17-25, 2017.
[Is] ISSN:0507-4088
[Cp] País de publicação:Russia (Federation)
[La] Idioma:eng
[Ab] Resumo:A task of creating a universal platform for engineering affordable recombinant producers of viral proteins conserving immunogenicity has not been solved yet. High toxicity of the viral proteins for the host cells, low yield and abnormal folding of the products often present severe obstacles to obtaining producers of the viral proteins. In this work, we report a new method of engineering and screening of deletion libraries from the viral antigen genes. This method allows selection of artificial derivatives of these genes adapted for expression in microbial producer cells. The method involves PCR amplification of the gene fragments using a system of randomized and adapter primers, which allows the spontaneous formation of duplexes from the random primers in the absence of the template DNA to be prevented. For selecting variants capable of in vivo expression, the obtained PCR products are cloned to a special vector of a direct phenotypical selection pQL30. It contains E. coli ß-galactosidase gene with an inserted polylinker producing a frame-shift mutation. Using this screening method, an artificial variant of hepatitis C (HCV) NS5a gene with optimal biotechnological properties was established. 27 clinical specimens of 1670 bp long HCV1b NS5a fragments were used as a source gene. A PCR bank of the deletion derivatives was produced. 40 LacZ-positive clones based on pQL30 vector with a 50-700 bp long insertion were selected. The LacZ activity of the cell lysates and the immunogenicity of the products were tested. As a result, a single clone encoding a soluble protein with Mr = 114 kDa was selected. Its yield reached 0.3% of the total cell protein. It was highly reactive with sera of HCV 1b infected patients but not with sera of the healthy donors.
[Mh] Termos MeSH primário: Antígenos Virais/genética
Escherichia coli/genética
Hepacivirus/genética
Hepatite C/diagnóstico
Engenharia de Proteínas/métodos
Proteínas Recombinantes/genética
Proteínas não Estruturais Virais/genética
[Mh] Termos MeSH secundário: Antígenos Virais/biossíntese
Antígenos Virais/imunologia
Antígenos Virais/isolamento & purificação
Proteínas de Bactérias/genética
Proteínas de Bactérias/metabolismo
Primers do DNA/síntese química
Primers do DNA/genética
Escherichia coli/metabolismo
Mutação da Fase de Leitura
Biblioteca Gênica
Vetores Genéticos/química
Vetores Genéticos/metabolismo
Hepacivirus/imunologia
Hepatite C/imunologia
Hepatite C/virologia
Seres Humanos
Soros Imunes/química
Óperon Lac
Proteínas Recombinantes/biossíntese
Proteínas Recombinantes/imunologia
Proteínas Recombinantes/isolamento & purificação
Proteínas não Estruturais Virais/biossíntese
Proteínas não Estruturais Virais/imunologia
Proteínas não Estruturais Virais/isolamento & purificação
beta-Galactosidase/genética
beta-Galactosidase/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antigens, Viral); 0 (Bacterial Proteins); 0 (DNA Primers); 0 (Immune Sera); 0 (NS-5 protein, hepatitis C virus); 0 (Recombinant Proteins); 0 (Viral Nonstructural Proteins); EC 3.2.1.23 (beta-Galactosidase)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180208
[Lr] Data última revisão:
180208
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180112
[St] Status:MEDLINE


  3 / 6777 MEDLINE  
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[PMID]:28807902
[Au] Autor:Li Y; Wang C; Cai W; Sengupta S; Zavortink M; Deng H; Girton J; Johansen J; Johansen KM
[Ad] Endereço:Roy J. Carver Department of Biochemistry, Biophysics, and Molecular Biology, Iowa State University, Ames, Iowa 50011, USA.
[Ti] Título:H2Av facilitates H3S10 phosphorylation but is not required for heat shock-induced chromatin decondensation or transcriptional elongation.
[So] Source:Development;144(18):3232-3240, 2017 09 15.
[Is] ISSN:1477-9129
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:A model has been proposed in which JIL-1 kinase-mediated H3S10 and H2Av phosphorylation is required for transcriptional elongation and heat shock-induced chromatin decondensation. However, here we show that although H3S10 phosphorylation is indeed compromised in the null mutant, chromatin decondensation at heat shock loci is unaffected in the absence of JIL-1 as well as of H2Av and that there is no discernable decrease in the elongating form of RNA polymerase II in either mutant. Furthermore, mRNA for the major heat shock protein Hsp70 is transcribed at robust levels in both and null mutants. Using a different chromatin remodeling paradigm that is JIL-1 dependent, we provide evidence that ectopic tethering of JIL-1 and subsequent H3S10 phosphorylation recruits PARP-1 to the remodeling site independently of H2Av phosphorylation. These data strongly suggest that H2Av or H3S10 phosphorylation by JIL-1 is not required for chromatin decondensation or transcriptional elongation in .
[Mh] Termos MeSH primário: Cromatina/metabolismo
Proteínas de Drosophila/metabolismo
Drosophila melanogaster/metabolismo
Histonas/metabolismo
Fosfosserina/metabolismo
Elongação da Transcrição Genética
[Mh] Termos MeSH secundário: Animais
Eucromatina/metabolismo
Proteínas de Fluorescência Verde/metabolismo
Resposta ao Choque Térmico/genética
Immunoblotting
Imuno-Histoquímica
Óperon Lac/genética
Mutação/genética
Fosforilação
Poli(ADP-Ribose) Polimerases/metabolismo
Cromossomos Politênicos/metabolismo
Proteínas Serina-Treonina Quinases/metabolismo
Transgenes
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, N.I.H., EXTRAMURAL
[Nm] Nome de substância:
0 (Chromatin); 0 (Drosophila Proteins); 0 (Euchromatin); 0 (H2AV protein, Drosophila); 0 (Histones); 147336-22-9 (Green Fluorescent Proteins); 17885-08-4 (Phosphoserine); EC 2.4.2.30 (Poly(ADP-ribose) Polymerases); EC 2.7.11.1 (JIL-1 protein, Drosophila); EC 2.7.11.1 (Protein-Serine-Threonine Kinases)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171126
[Lr] Data última revisão:
171126
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170816
[St] Status:MEDLINE
[do] DOI:10.1242/dev.151134


  4 / 6777 MEDLINE  
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[PMID]:28732013
[Au] Autor:Luo M; Yang S; Li X; Liu P; Xue J; Zhou X; Su K; Xu X; Qing Y; Qiu J; Li Y
[Ad] Endereço:School of Public Health and Management, Chongqing Medical University, Chongqing, China.
[Ti] Título:The KP1_4563 gene is regulated by the cAMP receptor protein and controls type 3 fimbrial function in Klebsiella pneumoniae NTUH-K2044.
[So] Source:PLoS One;12(7):e0180666, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Klebsiella pneumoniae (K. pneumoniae) is an opportunistic pathogen that can adhere to host cells or extracellular matrix via type 1 and type 3 fimbriae. KP1_4563 is a gene encoding a hypothetical protein in K. pneumoniae NTUH-K2044. KP1_4563 is located between the type 1 and type 3 fimbrial gene clusters and is likely associated with fimbrial function given its putative conserved domains of unknown function (DUF1471). Cyclic AMP receptor protein (CRP) regulates virulence-related gene expression and is a crucial transcriptional regulator in many bacteria. The predicted DNA recognition motif of CRP is present in the KP1_4563 promoter region. This study aimed to investigate the function of KP1_4563 in fimbriae and its transcriptional regulation mechanism by CRP. We generated Kp-Δ4563 mutant and complementation strains. We utilized phenotype and adhesion assays to evaluate the role of KP1_4563 in fimbriae. We conducted quantitative RT-PCR (qRT-PCR), LacZ fusion, electrophoretic mobility shift, and DNase I footprinting assays to study the transcriptional regulation of KP1_4563 gene by CRP. We found that KP1_4563 negatively regulates the function of type 3 fimbriae. Compared with NTUH-K2044, the absence of KP1_4563 enhanced the ability of Kp-Δ4563 to adhere to A549 cells. CRP negatively regulates KP1_4563 by directly binding to its promoter region. KP1_4563 plays an important role in type 3 fimbrial function. This novel insight will assist in the development of strategies for preventing K. pneumoniae infection.
[Mh] Termos MeSH primário: Proteína Receptora de AMP Cíclico/metabolismo
Proteínas de Fímbrias/metabolismo
Klebsiella pneumoniae/genética
Klebsiella pneumoniae/metabolismo
[Mh] Termos MeSH secundário: Aderência Bacteriana/fisiologia
Pegada de DNA
Desoxirribonuclease I/metabolismo
Ensaio de Desvio de Mobilidade Eletroforética
Escherichia coli
Regulação da Expressão Gênica/fisiologia
Testes de Hemaglutinação
Óperon Lac
Mananas/química
Fenótipo
Reação em Cadeia da Polimerase em Tempo Real
Saccharomyces cerevisiae
Deleção de Sequência
Transcrição Genética/fisiologia
beta-Galactosidase/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Cyclic AMP Receptor Protein); 0 (Mannans); 147680-16-8 (Fimbriae Proteins); EC 3.1.21.1 (Deoxyribonuclease I); EC 3.2.1.23 (beta-Galactosidase)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170927
[Lr] Data última revisão:
170927
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170722
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0180666


  5 / 6777 MEDLINE  
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[PMID]:28686609
[Au] Autor:Landman J; Brewster RC; Weinert FM; Phillips R; Kegel WK
[Ad] Endereço:Van 't Hoff Laboratory for Physical & Colloid Chemistry, Utrecht University, Utrecht, the Netherlands.
[Ti] Título:Self-consistent theory of transcriptional control in complex regulatory architectures.
[So] Source:PLoS One;12(7):e0179235, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Individual regulatory proteins are typically charged with the simultaneous regulation of a battery of different genes. As a result, when one of these proteins is limiting, competitive effects have a significant impact on the transcriptional response of the regulated genes. Here we present a general framework for the analysis of any generic regulatory architecture that accounts for the competitive effects of the regulatory environment by isolating these effects into an effective concentration parameter. These predictions are formulated using the grand-canonical ensemble of statistical mechanics and the fold-change in gene expression is predicted as a function of the number of transcription factors, the strength of interactions between the transcription factors and their DNA binding sites, and the effective concentration of the transcription factor. The effective concentration is set by the transcription factor interactions with competing binding sites within the cell and is determined self-consistently. Using this approach, we analyze regulatory architectures in the grand-canonical ensemble ranging from simple repression and simple activation to scenarios that include repression mediated by DNA looping of distal regulatory sites. It is demonstrated that all the canonical expressions previously derived in the case of an isolated, non-competing gene, can be generalised by a simple substitution to their grand canonical counterpart, which allows for simple intuitive incorporation of the influence of multiple competing transcription factor binding sites. As an example of the strength of this approach, we build on these results to present an analytical description of transcriptional regulation of the lac operon.
[Mh] Termos MeSH primário: Proteínas de Ligação a DNA/genética
Regulação Bacteriana da Expressão Gênica/genética
Redes Reguladoras de Genes/genética
Óperon Lac/genética
[Mh] Termos MeSH secundário: Sítios de Ligação
Proteínas de Ligação a DNA/biossíntese
Escherichia coli/genética
Modelos Teóricos
Ligação Proteica/genética
Fatores de Transcrição/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA-Binding Proteins); 0 (Transcription Factors)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171004
[Lr] Data última revisão:
171004
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170708
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0179235


  6 / 6777 MEDLINE  
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[PMID]:28663257
[Au] Autor:Sawada H; Rateri DL; Moorleghen JJ; Majesky MW; Daugherty A
[Ad] Endereço:From the Saha Cardiovascular Research Center (H.S., D.L.R., J.J.M., A.D.) and Department of Physiology (A.D.), University of Kentucky, Lexington; Seattle Children's Research Institute, Washington (M.W.M.); and Department of Pediatrics and Department of Pathology, University of Washington, Seattle (M
[Ti] Título:Smooth Muscle Cells Derived From Second Heart Field and Cardiac Neural Crest Reside in Spatially Distinct Domains in the Media of the Ascending Aorta-Brief Report.
[So] Source:Arterioscler Thromb Vasc Biol;37(9):1722-1726, 2017 Sep.
[Is] ISSN:1524-4636
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:OBJECTIVE: Smooth muscle cells (SMCs) of the proximal thoracic aorta are embryonically derived from the second heart field (SHF) and cardiac neural crest (CNC). However, distributions of these embryonic origins are not fully defined. The regional distribution of SMCs of different origins is speculated to cause region-specific aortopathies. Therefore, the aim of this study was to determine the distribution of SMCs of SHF and CNC origins in the proximal thoracic aorta. APPROACH AND RESULTS: Mice with repressed LacZ in the ROSA26 locus were bred to those expressing controlled by either the Wnt1 or Mef2c (myocyte-specific enhancer factor 2c) promoter to trace CNC- and SHF-derived SMCs, respectively. Thoracic aortas were harvested, and activity of ß-galactosidase was determined. Aortas from Wnt1- mice had ß-galactosidase-positive areas throughout the region from the proximal ascending aorta to just distal of the subclavian arterial branch. Unexpectedly, ß-galactosidase-positive areas in Mef2c- mice extended from the aortic root throughout the ascending aorta. This distribution occurred independent of sex and aging. Cross and sagittal aortic sections demonstrated that CNC-derived cells populated the inner medial aspect of the anterior region of the ascending aorta and transmurally in the media of the posterior region. Interestingly, outer medial cells throughout anterior and posterior ascending aortas were derived from the SHF. ß-Galactosidase-positive medial cells of both origins colocalized with an SMC marker, α-actin. CONCLUSIONS: Both CNC- and SHF-derived SMCs populate the media throughout the ascending aorta. The outer medial cells of the ascending aorta form a sleeve populated by SHF-derived SMCs.
[Mh] Termos MeSH primário: Linhagem da Célula
Coração/embriologia
Músculo Liso Vascular/fisiologia
Miocárdio
Miócitos de Músculo Liso/fisiologia
Crista Neural/fisiologia
Túnica Média/fisiologia
[Mh] Termos MeSH secundário: Fatores Etários
Animais
Aorta Torácica/embriologia
Aorta Torácica/metabolismo
Aorta Torácica/fisiologia
Feminino
Regulação da Expressão Gênica no Desenvolvimento
Genótipo
Integrases/genética
Óperon Lac
Fatores de Transcrição MEF2/genética
Masculino
Camundongos Transgênicos
Morfogênese
Músculo Liso Vascular/embriologia
Músculo Liso Vascular/metabolismo
Miocárdio/metabolismo
Miócitos de Músculo Liso/metabolismo
Crista Neural/embriologia
Crista Neural/metabolismo
Fenótipo
Regiões Promotoras Genéticas
RNA não Traduzido/genética
Fatores Sexuais
Túnica Média/embriologia
Túnica Média/metabolismo
Proteína Wnt1/genética
beta-Galactosidase/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Gt(ROSA)26Sor non-coding RNA, mouse); 0 (MEF2 Transcription Factors); 0 (Mef2c protein, mouse); 0 (RNA, Untranslated); 0 (Wnt1 Protein); 0 (Wnt1 protein, mouse); EC 2.7.7.- (Cre recombinase); EC 2.7.7.- (Integrases); EC 3.2.1.23 (beta-Galactosidase)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170906
[Lr] Data última revisão:
170906
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170701
[St] Status:MEDLINE
[do] DOI:10.1161/ATVBAHA.117.309599


  7 / 6777 MEDLINE  
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[PMID]:28494968
[Au] Autor:Zander D; Samaga D; Straube R; Bettenbrock K
[Ad] Endereço:Max-Planck-Institute for Dynamics of Complex Technical Systems, Magdeburg, Germany.
[Ti] Título:Bistability and Nonmonotonic Induction of the lac Operon in the Natural Lactose Uptake System.
[So] Source:Biophys J;112(9):1984-1996, 2017 May 09.
[Is] ISSN:1542-0086
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The Escherichia coli lac operon is regulated by a positive feedback loop whose potential to generate an all-or-none response in single cells has been a paradigm for bistable gene expression. However, so far bistable lac induction has only been observed using gratuitous inducers, raising the question about the biological relevance of bistable lac induction in the natural setting with lactose as the inducer. In fact, the existing experimental evidence points to a graded rather than an all-or-none response in the natural lactose uptake system. In contrast, predictions based on computational models of the lactose uptake pathway remain controversial. Although some argue in favor of bistability, others argue against it. Here, we reinvestigate lac operon expression in single cells using a combined experimental/modeling approach. To this end, we parameterize a well-supported mathematical model using transient measurements of LacZ activity upon induction with different amounts of lactose. The resulting model predicts a monostable induction curve for the wild-type system, but indicates that overexpression of the LacI repressor would drive the system into the bistable regime. Both predictions were confirmed experimentally supporting the view that the wild-type lac induction circuit generates a graded response rather than bistability. More interestingly, we find that the lac induction curve exhibits a pronounced maximum at intermediate lactose concentrations. Supported by our data, a model-based analysis suggests that the nonmonotonic response results from saturation of the LacI repressor at low inducer concentrations and dilution of Lac enzymes due to an increased growth rate beyond the saturation point. We speculate that the observed maximum in the lac expression level helps to save cellular resources by limiting Lac enzyme expression at high inducer concentrations.
[Mh] Termos MeSH primário: Proteínas de Escherichia coli/metabolismo
Escherichia coli/genética
Óperon Lac
Lactose/metabolismo
Modelos Biológicos
[Mh] Termos MeSH secundário: Meios de Cultura
Indução Enzimática
Escherichia coli/metabolismo
Proteínas de Escherichia coli/genética
Galactose/metabolismo
Regulação Bacteriana da Expressão Gênica
Glucose/metabolismo
Microscopia de Fluorescência
Ácido Succínico/metabolismo
beta-Galactosidase/biossíntese
beta-Galactosidase/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Culture Media); 0 (Escherichia coli Proteins); AB6MNQ6J6L (Succinic Acid); EC 3.2.1.23 (beta-Galactosidase); IY9XDZ35W2 (Glucose); J2B2A4N98G (Lactose); X2RN3Q8DNE (Galactose)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170808
[Lr] Data última revisão:
170808
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170513
[St] Status:MEDLINE


  8 / 6777 MEDLINE  
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[PMID]:28475610
[Au] Autor:Shen X; Bao W; Yu W; Liang R; Nguyen B; Liu Y
[Ad] Endereço:Department of Biology and Biochemistry, University of Houston, Houston, TX, United States of America.
[Ti] Título:An improved method with high sensitivity and low background in detecting low ß-galactosidase expression in mouse embryos.
[So] Source:PLoS One;12(5):e0176915, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:LacZ is widely used as a reporter in studies of gene expression patterns. ß-galactosidase, the product of LacZ gene, is usually detected by X-gal/FeCN staining. In X-gal/FeCN staining, ß-galactosidase catalyzes X-gal to produce blue precipitates, which indicate the expression patterns of the gene of interest. A newer LacZ detection method using S-gal/TNBT is more sensitive but plagued by high background. Here, we describe an improved procedure that combines advantageous steps from the two methods. By comparing with X-gal/FeCN and S-gal/TNBT methods in detecting the expression patterns of miR-322/503 and miR-451 at a series of developmental stages, the improved method showed higher sensitivity and lower background. Thus, the improved method could be an alternative way of ß-galactosidase staining in low gene expression situations.
[Mh] Termos MeSH primário: Embrião de Mamíferos/enzimologia
beta-Galactosidase/metabolismo
[Mh] Termos MeSH secundário: Animais
Feminino
Óperon Lac
Limite de Detecção
Masculino
Camundongos
Camundongos Knockout
Reprodutibilidade dos Testes
beta-Galactosidase/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
EC 3.2.1.23 (beta-Galactosidase)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170915
[Lr] Data última revisão:
170915
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170506
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0176915


  9 / 6777 MEDLINE  
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[PMID]:28415454
[Au] Autor:Rasoulianboroujeni M; Kupgan G; Moghadam F; Tahriri M; Boughdachi A; Khoshkenar P; Ambrose JJ; Kiaie N; Vashaee D; Ramsey JD; Tayebi L
[Ad] Endereço:Marquette University School of Dentistry, Milwaukee, WI 53233, USA.
[Ti] Título:Development of a DNA-liposome complex for gene delivery applications.
[So] Source:Mater Sci Eng C Mater Biol Appl;75:191-197, 2017 Jun 01.
[Is] ISSN:1873-0191
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:The association structures formed by cationic liposomes and DNA (Deoxyribonucleic acid)-liposome have been effectively utilized as gene carriers in transfection assays. In this research study, cationic liposomes were prepared using a modified lipid film hydration method consisting of a lyophilization step for gene delivery applications. The obtained results demonstrated that the mean particle size had no significant change while the polydispersity (PDI) increased after lyophilization. The mean particle size slightly reduced after lyophilization (520±12nm to 464±25nm) while the PDI increased after lyophilization (0.094±0.017 to 0.220±0.004). In addition. The mean particle size of vesicles increases when DNA is incorporated to the liposomes (673±27nm). According to the Scanning Electron Microscopy (SEM) and transmission electron microscopy (TEM) images, the spherical shape of liposomes confirmed their successful preservation and reconstitution from the powder. It was found that liposomal formulation has enhanced transfection considerably compared to the naked DNA as negative control. Finally, liposomal formulation in this research had a better function than Lipofectamine® 2000 as a commercialized product because the cellular activity (cellular protein) was higher in the prepared lipoplex than Lipofectamine® 2000.
[Mh] Termos MeSH primário: DNA
Técnicas de Transferência de Genes
[Mh] Termos MeSH secundário: DNA/química
DNA/genética
DNA/farmacologia
Células HEK293
Seres Humanos
Óperon Lac
Lipossomos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Liposomes); 9007-49-2 (DNA)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:170724
[Lr] Data última revisão:
170724
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170419
[St] Status:MEDLINE


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[PMID]:28336461
[Au] Autor:Toulouse C; Häse CC; Steuber J
[Ad] Endereço:Institute of Microbiology, University of Hohenheim (Stuttgart), 70599 Stuttgart, Germany.
[Ti] Título:Chloroform-free permeabilization for improved detection of ß-galactosidase activity in Vibrio cholerae.
[So] Source:J Microbiol Methods;137:1-2, 2017 Jun.
[Is] ISSN:1872-8359
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:LacZ (ß-galactosidase) is used to monitor the transcription of genes in reporter strains carrying the lacZ gene under the control of a promotor of interest. This protocol for LacZ activity determinations in Vibrio cholerae following detergent lysis results in 2.5-fold increase of LacZ activities compared to lysis with chloroform.
[Mh] Termos MeSH primário: Clorofórmio/farmacologia
Vibrio cholerae/efeitos dos fármacos
Vibrio cholerae/enzimologia
beta-Galactosidase/metabolismo
[Mh] Termos MeSH secundário: Proteínas de Bactérias/genética
Membrana Celular/metabolismo
Detergentes/farmacologia
Genes Reporter
Óperon Lac
Nitrofenóis/metabolismo
Regiões Promotoras Genéticas
Inibidores de Proteases/farmacologia
Transcrição Genética
Vibrio cholerae/genética
beta-Galactosidase/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Bacterial Proteins); 0 (Detergents); 0 (Nitrophenols); 0 (Protease Inhibitors); 7V31YC746X (Chloroform); EC 3.2.1.23 (beta-Galactosidase)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170808
[Lr] Data última revisão:
170808
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170325
[St] Status:MEDLINE



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