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Pesquisa : D12.644.770 [Categoria DeCS]
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Texto completo SciELO Brasil
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Id: lil-709455
Autor: Feng, Bao Zhen; Li, Peiqian.
Título: Cloning, characterization and expression of a novel laccase gene Pclac2 from Phytophthora capsici
Fonte: Braz. j. microbiol;45(1):351-358, 2014. ilus.
Idioma: en.
Projeto: Program of University of Science and Technology of Shanxi Province; . Natural Science Foundation for Young Scientists of Shanxi Province. 2013021024-6; . Yuncheng University.
Resumo: Laccases are blue copper oxidases (E.C. 1.10.3.2) that catalyze the one-electron oxidation of phenolics, aromatic amines, and other electron-rich substrates with the concomitant reduction of O2 to H2O. A novel laccase gene pclac2 and its corresponding full-length cDNA were cloned and characterized from Phytophthora capsici for the first time. The 1683 bp full-length cDNA of pclac2 encoded a mature laccase protein containing 560 amino acids preceded by a signal peptide of 23 amino acids. The deduced protein sequence of PCLAC2 showed high similarity with other known fungal laccases and contained four copper-binding conserved domains of typical laccase protein. In order to achieve a high level secretion and full activity expression of PCLAC2, expression vector pPIC9K with the Pichia pastoris expression system was used. The recombinant PCLAC2 protein was purified and showed on SDS-PAGE as a single band with an apparent molecular weight ca. 68 kDa. The high activity of purified PCLAC2, 84 U/mL, at the seventh day induced with methanol, was observed with 2,2'-azino-di-(3-ethylbenzothialozin-6-sulfonic acid) (ABTS) as substrate. The optimum pH and temperature for ABTS were 4.0 and 30 ºC, respectively . The reported data add a new piece to the knowledge about P. Capsici laccase multigene family and shed light on potential function about biotechnological and industrial applications of the individual laccase isoforms in oomycetes.
Descritores: Lacase/genética
Lacase/metabolismo
Phytophthora/enzimologia
-Clonagem Molecular
Sequência Conservada
Estabilidade Enzimática
Expressão Gênica
Concentração de Íons de Hidrogênio
Lacase/química
Lacase/isolamento & purificação
Peso Molecular
Fases de Leitura Aberta
Estrutura Terciária de Proteína
Phytophthora/genética
Pichia/genética
Sinais Direcionadores de Proteínas/genética
Proteínas Recombinantes/química
Proteínas Recombinantes/genética
Proteínas Recombinantes/isolamento & purificação
Proteínas Recombinantes/metabolismo
Homologia de Sequência de Aminoácidos
Temperatura Ambiente
Tipo de Publ: Research Support, Non-U.S. Gov't
Responsável: BR1.1 - BIREME


  2 / 7 LILACS  
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Texto completo SciELO Chile
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Id: lil-591914
Autor: Naseer Cheema, Hafiza Masooma; Bashir, Aftab; Khatoon, Asia; Iqbal, Nadia; Zafar, Yusuf; Malik, Kauser A.
Título: Molecular characterization and transcriptome profiling of expansin genes isolated from Calotropis procera fibers
Fonte: Electron. j. biotechnol;13(6):10-11, Nov. 2010. ilus, tab.
Idioma: en.
Resumo: The Calotropis procera seed fibers provide an excellent model system to study the genes involved in fiber elongation, fineness and strength. Expansins constitute one of the important gene families involved in plant cell expansion and other cell wall modification processes. Four homologs of Expansin A gene i.e. CpEXPA1, CpEXPA2, CpEXPA3 and CpEXPA4 were isolated from the cDNA library obtained from fast growing Calotropis procera fibers. These homologs represented typical Expansin A family. Each of them had two conserved domains including GH45 like domain and the putative polysaccharide binding domain. The deduced amino acid sequences of the homologs indicated three conserved motifs: i) eight cysteine residues at N-terminus, ii) four tryptophan residues at C-terminus and iii) a Histidine-Phenylalanine-Aspartate motif in the center of the sequence. The presence of N-terminal signal peptide consisting of hydrophobic amino acids and a transmembrane region in all these expansin isoforms suggests their cotranslational insertion into the endoplasmic reticulum and then transportation to the cell wall by secretory pathway. The relative quantification of the four expansins in root, stem, fiber and leave tissues indicated that the transcripts of CpEXPA1, CpEXPA2, CpEXPA3 and CpEXPA4 are variably transcribed in these tissues. The lowest transcription of all the four Expansin A isoforms was observed in elongating roots indicating that root tissue might be having specific expansins other than those confined to air grown organs.
Descritores: Fibra de Algodão
Calotropis/genética
Calotropis/química
Proteínas de Plantas/genética
-DNA Complementar
Perfilação da Expressão Gênica
Genes de Plantas
Filogenia
Sinais Direcionadores de Proteínas
Proteínas de Plantas/química
RNA Mensageiro
Reação em Cadeia da Polimerase/métodos
Análise de Sequência
Responsável: CL1.1 - Biblioteca Central


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Texto completo SciELO Brasil
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Id: lil-416283
Autor: Xinmin, Li; Kim, Jaejung; Zhou, Jian; Gu, Weikuan; Quigg, Richard.
Título: Use of signal thresholds to determine significant changes in microarray data analyses
Fonte: Genet. mol. biol;28(2):191-200, 2005. tab, graf.
Idioma: en.
Resumo: The use of a constant fold-change to determine significant changes in gene expression has been widely accepted for its intuition and ease of use in microarray data analysis, but this concept has been increasingly criticized because it does not reflect signal intensity and can result in a substantial number of false positives and false negatives. To resolve this dilemma, we have analyzed 65 replicate Affymetrix chip-chip comparisons and determined a series of user adjustable signal-dependent thresholds which do not require replicates and offer a 95 percent confidence interval. Quantitative RT-PCR shows that such thresholds significantly improve the power to discriminate biological changes in mRNA from noise and reduce false calls compared to the traditional two-fold threshold. The user-friendly nature of this approach means that it can be easily applied by any user of microarray analysis, even those without any specialized knowledge of computational techniques or statistics. Noise is a function of signal intensity not only for Affymetrix data but also for cDNA array data, analysis of which may also be benefited by our methodology.
Descritores: Análise de Sequência com Séries de Oligonucleotídeos
Sinais Direcionadores de Proteínas
Estatística como Assunto
Limites: Seres Humanos
Responsável: BR1.1 - BIREME


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Id: lil-335981
Autor: Burdick, Debra; Le Gall, Annick H; Rodriguez-Boulan, Enrique.
Título: Vesicular transport: implications for cell polarity
Fonte: Biocell;20(3):343-353, Dec. 1996.
Idioma: en.
Resumo: In polarized cells intracellular sorting of plasma membrane proteins occurs to a large extent at the trans-Golgi network, giving rise to vesicles destined for distinct plasma membrane domains. This review discusses the several pathways, both direct and indirect, which lead to protein incorporation into the correct cell surface, as well as the mechanisms involved. Proteins contain signals which direct their incorporation into the distinct vesicles destined for plasma membrane microdomains. Specific coat proteins are involved in vesicle assembly and are likely to play a role in the generation of discrete vesicle populations. Molecules involved in vesicle docking and fusion may also add specificity to the targeting process.
Descritores: Polaridade Celular
Proteínas de Membrana/metabolismo
Vesículas Revestidas/fisiologia
-Sequência de Aminoácidos
Transporte Biológico
Linhagem Celular
Complexo de Golgi
Rim
Fusão de Membrana
Modelos Biológicos
Dados de Sequência Molecular
Organelas
Sinais Direcionadores de Proteínas
Proteínas de Membrana/classificação
Proteínas de Membrana/fisiologia
Tirosina
Limites: Animais
Cães
Tipo de Publ: Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.
Responsável: BR1.1 - BIREME


  5 / 7 LILACS  
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Texto completo SciELO Brasil
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Id: lil-325930
Autor: Surpili, Marcelo J; Müller-Röber, Bernd; Willmitzer, Lothar.
Título: A yeast-based model system for cloning secreted and membrane proteins
Fonte: An. acad. bras. ciênc;74(4):599-608, Dec. 2002. ilus, graf.
Idioma: en.
Resumo: The targeting of proteins to cell organelles and membranes, or of proteins destined to secretion, is coordinated by signal sequences located at the 5´-end of their respective genes. A signal sequence trap system was envisaged in which a truncated version of the yeast acid phosphatase pho5 gene lacking the start codon and signal sequence could serve as a reporter gene. A fraction enriched in 5´-end fragments obtained by PCR from a potato guard-cell cDNA library was cloned in frame to the acid phosphatase gene and the acid phosphatase activity was assayed directly in yeast colonies grown on selective medium. Putative signal sequences targeting the acid phosphatase to the membrane or to the outside of the cell were used to screen the cDNA bank in order to recover the original full-size sequence which gave rise to the signal sequence. Two unknown sequences displaying marked tissue-specific expression were retrieved, one of them (YE139) with a higher expression level in green buds and stem cells, and the other one (YE290) with a higher expression level in androceum, gyneceum, and roots. The limitations of the system are further analyzed using other sequences as control
Descritores: Fosfatase Ácida
Clonagem Molecular
Proteínas de Membrana
Sinais Direcionadores de Proteínas
Saccharomyces cerevisiae
-Fosfatase Ácida
Sequência de Bases
Biblioteca Gênica
Proteínas de Membrana
Modelos Biológicos
Reação em Cadeia da Polimerase
Canais de Potássio
Sinais Direcionadores de Proteínas
Transdução de Sinais
Solanum tuberosum
Tipo de Publ: Research Support, Non-U.S. Gov't
Responsável: BR1.1 - BIREME


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Texto completo SciELO Chile
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Id: lil-301906
Autor: Martínez T., Alejandra; González Correa, Carlos; Kawaguchi P., Fernando; Montoya, Rolando; Corvalán, Alejandro; Madariaga B., Jaime; Roa S., Jorge; García C., Apolinaria; Salgado S., Fernando; Solar, Henry; Palma, Mariana.
Título: Helicobacter pylori: análisis de cagA y genotipificación de vacA en Chile: detección de una cepa s2/m1 / Helicobacter pylori: cagA status and vacA genotyping in Chile, detection of a s2/m1 strain
Fonte: Rev. méd. Chile;129(10):1147-1153, oct. 2001. tab, graf.
Idioma: es.
Resumo: Background: The genes cagA and vacA encode H pylori virulence factors. Aim: To genotype these genes in H pylori strains isolated from patients with upper gastrointestinal symptoms. Material and methods: We studied 50 patients who underwent an upper gastrointestinal endoscopy, with positive culture for H pylori. Detection of cagA and vacA gerotyping was done using polymerase chain reactions. Results: The gene cagA was detected in 19 samples (38 per cent). Signal sequences s1 and s2 of vacA gene were detected in 16 samples each (32 per cent). There was simultaneous amplification of s1 and s2 in 6 samples and they were not detected in 9 samples. The middle region of vacA was m1 in 9 samples, m2 in 29 samples and there was simultaneous amplification of m1 and m2 in 12 samples. In 16 samples (32 per cent), more than one type of signal sequence or medial region was detected. Of those patients in whom vacA was the only genotype detected, 15 were s2/m2, 7 were s1/m1, 4 were s1/m2 and 1 was s2/m1. Conclusions: In these patients, the infection with cagA- H pylori strains, predominates, the prevalence of infection with s1 or s2 strains is similar and the predominant medial region is m2
Descritores: Genótipo
Helicobacter pylori
-Biópsia
Duodenoscopia
Eletroforese em Gel de Ágar
Gastroscopia
Sinais Direcionadores de Proteínas
Limites: Seres Humanos
Tipo de Publ: Revisão
Responsável: CL1.1 - Biblioteca Central


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Id: lil-252940
Autor: Anon.
Título: Premio Nobel en Medicina 1999 / Nobel Premiun in Medicine 1999
Fonte: Antioxid. calid. vida;6(25):26-7, sept. 1999. ilus.
Idioma: es.
Descritores: Medicina
Prêmio Nobel
Sinais Direcionadores de Proteínas/fisiologia
-Partícula de Reconhecimento de Sinal/isolamento & purificação
Partícula de Reconhecimento de Sinal/fisiologia
Sinais Direcionadores de Proteínas/isolamento & purificação
Sinais Direcionadores de Proteínas/metabolismo
Limites: Seres Humanos
Responsável: AR144.1 - CIBCHACO - Centro de Información Biomedica del Chaco



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