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  1 / 21 LILACS  
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Id: biblio-974318
Autor: Andreolla, Ana Paula; Erpen, Luana Marina Scheer; Frandoloso, Rafael; Kreutz, Luiz Carlos.
Título: Development of an indirect ELISA based on recombinant capsid protein to detect antibodies to bovine leukemia virus
Fonte: Braz. j. microbiol;49(supl.1):68-75, 2018. tab, graf.
Idioma: en.
Projeto: CAPES; . SDECT; . CNPq.
Resumo: Abstract Serological testing and culling infected animals are key management practices aiming eradication of bovine leukemia virus infection. Here, we report the development of an indirect ELISA based on BLV recombinant capsid protein (BLVp24r) to detect anti-BLV antibodies in cattle serum. The BLVp24r was expressed in Escherichia coli and purified by affinity chromatography, and then used to set up the ELISA parameters. The Polysorp ® plate coated with 50 ng of antigen/well and bovine serum diluted 1:100 gave the best results during standardization. Using sera from infected and non-infected cattle we set up the cutoff point at 0.320 (OD450 nm) with a sensitivity of 98.5% and specificity of 100.0%. Then, we tested 1.187 serum samples from dairy (736 samples) and beef cattle (451 samples) with unknown status to BLV. We found that 31.1% (229/736) and 9.5% (43/451) of samples amongst dairy and beef cattle, respectively, had IgGs to BLV. The rate of agreement with a commercial competitive ELISA was 84.3% with a κ value of 0.68. Thus, our BLVp24r iELISA is suitable to detect BLV infected animals and should be a useful tool to control BLV infection in cattle.
Descritores: Ensaio de Imunoadsorção Enzimática/métodos
Testes Sorológicos/métodos
Leucose Enzoótica Bovina/diagnóstico
Vírus da Leucemia Bovina/imunologia
Proteínas do Capsídeo/imunologia
Anticorpos Antivirais/sangue
-Proteínas Recombinantes/análise
Proteínas Recombinantes/genética
Proteínas Recombinantes/imunologia
Ensaio de Imunoadsorção Enzimática/instrumentação
Sensibilidade e Especificidade
Leucose Enzoótica Bovina/sangue
Leucose Enzoótica Bovina/virologia
Vírus da Leucemia Bovina/isolamento & purificação
Vírus da Leucemia Bovina/genética
Proteínas do Capsídeo/análise
Proteínas do Capsídeo/genética
Limites: Animais
Bovinos
Responsável: BR1.1 - BIREME


  2 / 21 LILACS  
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Id: biblio-974285
Autor: Pereira, Joylson de Jesus; Baumworcel, Natasha; Fioretti, Júlia Monassa; Domingues, Cinthya Fonseca; Moraes, Laís Fernandes de; Marinho, Robson dos Santos Souza; Vieira, Maria Clara Rodrigues; Pinto, Ana Maria Viana; de Castro, Tatiana Xavier.
Título: Molecular characterization of feline calicivirus variants from multicat household and public animal shelter in Rio de Janeiro, Brazil
Fonte: Braz. j. microbiol;49(4):777-784, Oct.-Dec. 2018. tab, graf.
Idioma: en.
Projeto: FAPERJ.
Resumo: ABSTRACT The aim of this study was to perform the molecular characterization of conserved and variable regions of feline calicivirus capsid genome in order to investigate the molecular diversity of variants in Brazilian cat population. Twenty-six conjunctival samples from cats living in five public short-term animal shelters and three multicat life-long households were analyzed. Fifteen cats had conjunctivitis, three had oral ulceration, eight had respiratory signs (cough, sneeze and nasal discharge) and nine were asymptomatic. Feline calicivirus were isolated in CRFK cells and characterized by reverse transcription PCR target to both conserved and variable regions of open reading frame 2. The amplicons obtained were sequenced. A phylogenetic analysis along with most of the prototypes available in GenBank database and an amino acid analysis were performed. Phylogenetic analysis based on both conserved and variable region revealed two clusters with an aLTR value of 1.00 and 0.98 respectively and the variants from this study belong to feline calicivirus genogroup I. No association between geographical distribution and/or clinical signs and clustering in phylogenetic tree was observed. The variants circulating in public short-term animal shelter demonstrated a high variability because of the relatively rapid turnover of carrier cats constantly introduced of multiple viruses into this location over time.
Descritores: Doenças do Gato/virologia
Calicivirus Felino/isolamento & purificação
Calicivirus Felino/genética
Infecções por Caliciviridae/veterinária
Animais de Estimação/virologia
-Filogenia
Brasil
Fases de Leitura Aberta
Genoma Viral
Calicivirus Felino/classificação
Infecções por Caliciviridae/virologia
Proteínas do Capsídeo/genética
Limites: Animais
Gatos
Responsável: BR1.1 - BIREME


  3 / 21 LILACS  
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Mores, Nelson
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Id: biblio-889245
Autor: Gava, Danielle; Serrão, Vitor Hugo Balasco; Fernandes, Lana Teixeira; Cantão, Mauricio Egídio; Ciacci-Zanella, Janice Reis; Morés, Nelson; Schaefer, Rejane.
Título: Structure analysis of capsid protein of Porcine circovirus type 2 from pigs with systemic disease
Fonte: Braz. j. microbiol;49(2):351-357, Apr.-June 2018. graf.
Idioma: en.
Projeto: Brazilian Agricultural Research Corporation-EMBRAPA; . São Paulo Research Foundation-FAPESP.
Resumo: Abstract Economic losses with high mortality rate associated with Porcine circovirus type 2 (PCV2) is reported worldwide. PCV2 commercial vaccine was introduced in 2006 in U.S. and in 2008 in Brazil. Although PCV2 vaccines have been widely used, cases of PCV2 systemic disease have been reported in the last years. Eleven nursery or fattening pigs suffering from PCV2 systemic disease were selected from eight PCV2-vaccinated farms with historical records of PCV2 systemic disease in Southern Brazil. PCV2 genomes were amplified and sequenced from lymph node samples of selected pigs. The comparison among the ORF2 amino acid sequences of PCV2 isolates revealed three amino acid substitutions in the positions F57I, N178S and A190T, respectively. Using molecular modeling, a structural model for the capsid protein of PCV2 was built. Afterwards, the mutated residues positions were identified in the model. The structural analysis of the mutated residues showed that the external residue 190 is close to an important predicted region for antibodies recognition. Therefore, changes in the viral protein conformation might lead to an inefficient antibody binding and this could be a relevant mechanism underlying the recent vaccine failures observed in swine farms in Brazil.
Descritores: Circovirus/química
Proteínas do Capsídeo/química
-Conformação Proteica
Suínos
Doenças dos Suínos/virologia
Brasil
Modelos Moleculares
Circovirus/isolamento & purificação
Circovirus/genética
Infecções por Circoviridae/veterinária
Infecções por Circoviridae/virologia
Substituição de Aminoácidos
Proteínas do Capsídeo/genética
Limites: Animais
Responsável: BR1.1 - BIREME


  4 / 21 LILACS  
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Id: biblio-1022119
Autor: Luchs, A; Timenetsky, M do C S T.
Título: Phylogenetic analysis of human group C rotavirus circulating in Brazil reveals a potential unique NSP4 genetic variant and high similarity with Asian strains
Fonte: Mol Genet Genomics;290(3):969-986, 2015.
Idioma: en.
Resumo: Group C rotaviruses (RVC) cause gastroenteritis in humans and animals worldwide, and the evidence for a possible zoonotic role has been recently provided. To gain information on the genetic diversity and relationships between human and animal RVC, we sequenced the VP4, VP7, and NSP4 genes of 12, 19, and 15 human strains, respectively, detected in São Paulo state during historical (1988 and 1993) and recent (2007 and 2008) Brazilian rotavirus surveillance. All RVC strains analyzed in the present study grouped into human genotype (G4-P[2]-E2), and did not show any evidence of animal ancestry. Phylogenetic analysis showed that RVC samples detected in 1988 and 1993 clustered together with strains from distinct continents, indicating that historical RVC strains circulating in São Paulo were closely related to those strains circulating worldwide. All three genes (VP7, VP4 and NSP4) of São Paulo RVC strains isolated in 2007-2008 exhibited close phylogenetic relationship with human RVC strains isolated in China and Japan, suggesting that they are genetically linked, and that a gene flow could be occurring between this Asian countries and Brazil. We identified two distinct clusters in the NSP4 phylogenetic tree. One cluster formed exclusively by human Brazilian strains detected in 1997 and 2003-2004 in Rio de Janeiro, Bahia, and Rio Grande do Sul states (Subgroup II) previously described in a different study, that displayed low sequence identities to other human strains formerly published, and to the Brazilian RVC strains (Subgroup I) characterized in the present study. These data suggests the circulation of two genetic profiles of the NSP4 gene in Brazil. High sequence diversity in NSP4 gene was previously reported in Asia, and additional diversity in NSP4 RVC strains spreading in the world should be expected. More in-depth molecular and epidemiological analysis of human RVC throughout the world will be needed to understand their diversity and clarify their evolution, as well as to develop classifications schemes.
Descritores:
Filogenia
Infecções por Rotavirus/virologia
Toxinas Biológicas/genética
Variação Genética
Brasil/epidemiologia
Humanos
RNA
RNA Viral/isolamento & purificação
Dados de Sequência Molecular
Sequência de Bases
Glicoproteínas
Glicoproteínas/genética
Glicoproteínas/química
Criança
Pré-Escolar
Demografia
Alinhamento de Sequência
Adolescente
Sequência de Aminoácidos
Proteínas não Estruturais Virais
Proteínas não Estruturais Virais/genética
Homologia de Sequência de Aminoácidos
Análise de Sequência de DNA
Rotavirus
Adulto
Proteínas do Capsídeo/química
Gastroenterite/virologia
Genótipo
Lactente
Animais
Pessoa de Meia-Idade
Antígenos Virais/genética
Responsável: BR91.2 - Centro de Documentação


  5 / 21 LILACS  
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Id: biblio-964569
Autor: Rodríguez-Martínez, Douglas; Figueira, Antonia dos Reis; Duarte, Priscilla de Sousa Geraldino; Costa, Suellen Bárbara Ferreira Galvino; González Olmedo, Justo.
Título: First report and molecular characterization of an isolate of papaya ringspot virus (PRSV-W) detected in pumpkin in Cuba / Primeiro relato e caracterização molecular de um isolado de papaya ringspot virus (PRSV-W) detectado em planta de abóbora proveniente de Cuba
Fonte: Biosci. j. (Online);31(4):1133-1142, july/aug. 2015.
Idioma: en.
Resumo: In this work, a virus isolate collected from pumpkin plants (Cucurbita pepo L.), showing severe symptoms of mosaic and leaf deformation, grown in Cuba, was analyzed using indicator plants, electron microscopy, and phylogenetic analysis. Plants of pumpkin, cv. Caserta, inoculated with this virus isolate showed mosaic, leaf distortion and blistering symptoms, whereas papaya plants were immune and did not show any symptoms. A transmission electron microscopic examination of leaf dip preparations made from infected pumpkin leaves revealed the presence of elongated and flexuous particles, approximately 780-800 x 12 nm in size. Genomic fragments containing the coat protein (CP) and HC-Pro genes, amplified by specific primers for Papaya ringspot virus, W strain (PRSV-W), showed amino acid identities of both genes higher than 94% when compared to other PRSV-W isolates from America. In the phylogenetic tree, this virus isolate has grouped with other virus isolates from America, Australia, and India and was more distant from the Asian isolates. Taken together, the analyses allow the conclusion that this virus isolate is a W strain of PRSV, detected for the first time in Cuba.

Neste trabalho um isolado viral coletado em planta de abóbora (Cucurbita pepo L), apresentando sintomas severos de mosaico e deformação foliar, proveniente de uma lavoura localizada em Cuba, foi analisado utilizando plantas indicadoras, microscopia eletrônica de transmissão e análise filogenética. Plantas de abóbora cv. Caserta, inoculadas com este isolado do vírus mostrou mosaico, distorção foliar e bolhas, enquanto que as plantas de mamão foram imunes e não apresentaram sintoma. Exame ao microscópio eletrônico de tranmissão de telas preparadas com a técnica leaf dip, empregando o extrato de folhas de abóbora infectadas revelou a presença de partículas alongadas e flexuosas, medindo cerca de 780-800 x 12 nm. A análise de fragmentos genômicos contendo os genes da proteína capsidial (CP) e HC-Pro, amplificados por primers específicos para Papaya ringspot virus, estirpe W (PRSV -W), mostrou identidades de aminoácidos superiores a 94 % quando ambos os genes foram comparados a outros isolados americanos de PRSV -W. Na árvore filogenética, este isolado estudado se agrupou com os isolados de PRSV-W da América, Austrália e Índia, ficando mais distante dos isolados asiáticos. Tomadas em conjunto, as análises permitem concluir que este isolado viral pertence à estirpe W do PRSV, detectada pela primeira vez em Cuba.
Descritores: Filogenia
Cucurbita pepo
Cuba
Cucurbita
Proteínas do Capsídeo
Biologia Molecular
Responsável: BR396.1 - Biblioteca Central


  6 / 21 LILACS  
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Id: lil-741274
Autor: Ali, Asad; Hussain, Adil; Ahmad, Musharaf.
Título: Occurrence and molecular characterization of Cucumber green mottle mosaic virus in cucurbit crops of KPK, Pakistan
Fonte: Braz. j. microbiol;45(4):1247-1253, Oct.-Dec. 2014. ilus, tab.
Idioma: en.
Resumo: Field survey of the cucurbit crops revealed a high incidence of Cucumber green mottle mosaic virus (CGMMV) in Khyber Pakhtunkhwa Province (KPK), Pakistan. Among the seven districts surveyed, average percent incidence of CGMMV was recorded up to 58.1% in district Nowshera, followed by 51.1% in district Charsada, 40.5% in district Swabi and 37.3% in district Mardan. In Swat and Dir districts average incidence CGMMV was recorded upto 31.2% and 29.4%, respectively. Among the different crops highest incidence in plain areas of KPK was recorded in bottle gourd (59.3%) followed by 56.3% in Squash, 54.5% in Pumpkin, 45.5% in Melon, 41.7% in Cucumber and 29.9% in Sponge gourd. In Northern hilly areas highest incidence of CGMMV (52.9%) was observed in pumpkin, followed by 49.6% in bottle gourd, 47.3% in squash, 45.1% in Melon 42.3% in cucumber and 41.6% in sponge gourd. Little variability was observed in the coat protein amino acid sequence identities of CGMMV Pakistan isolate, when compared with other reported isolates.
Descritores: Cucurbitaceae/virologia
Doenças das Plantas/virologia
Tobamovirus/isolamento & purificação
-Análise por Conglomerados
Proteínas do Capsídeo/genética
Variação Genética
Incidência
Dados de Sequência Molecular
Paquistão
Filogenia
Análise de Sequência de DNA
Homologia de Sequência de Aminoácidos
Tobamovirus/classificação
Tobamovirus/genética
Responsável: BR1.1 - BIREME


  7 / 21 LILACS  
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Texto completo SciELO Venezuela
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Id: lil-740332
Autor: Odreman-Macchioli, María; Vielma, Silvana; Atchley, Daniel; Comach, Guillermo; Ramirez, Alvaro; Pérez, Saberio; Téllez, Luis; Quintero, Beatriz; Hernández, Erick; Muñoz, Maritza; Mendoza, José.
Título: Analysis of real time PCR amplification efficiencies from three genomic region of dengue virus / Análisis de las eficiencias de amplificación por PCR en tiempo real de tres regiones genómicas del virus dengue
Fonte: Invest. clín;54(1):5-19, mar. 2013. tab.
Idioma: en.
Resumo: Early diagnosis of dengue virus (DENV) infection represents a key factor in preventing clinical complications attributed to the disease. The aim of this study was to evaluate the amplification efficiencies of an in-house quantitative real time-PCR (qPCR) assay of DENV, using the non-structural conserved genomic region protein-5 (NS5) versus two genomic regions usually employed for virus detection, the capsid/pre-membrane region (C-prM) and the 3'-noncoding region (3'NC). One-hundred sixty seven acute phase serum samples from febrile patients were used for validation purposes. Results showed that the three genomic regions had similar amplification profiles and correlation coefficients (0.987-0.999). When isolated viruses were used, the NS5 region had the highest qPCR efficiencies for the four serotypes (98-100%). Amplification from acute serum samples showed that 41.1% (67/167) were positive for the universal assay by at least two of the selected genomic regions. The agreement rates between NS5/C-prM and NS5/3'NC regions were 56.7% and 97%, respectively. Amplification concordance values between C-prM/NS5 and NS5/3'NC regions showed a weak (k= 0.109; CI 95%) and a moderate (k= 0.489; CI 95%) efficiencies in amplification, respectively. Serotyping assay using a singleplex NS5-TaqMan® format was much more sensitive than the C-prM/SYBR Green® I protocol (76%). External evaluation showed a high sensitivity (100%), specificity (78%) and high agreement between the assays. According to the results, the NS5 genomic region provides the best genomic region for optimal detection and typification of DENV in clinical samples.

El diagnóstico precoz de la infección por el virus dengue (DENV) constituye un elemento clave para la prevención de las complicaciones clínicas propias de la enfermedad. El objetivo del estudio fue evaluar la detección de DENV mediante un ensayo cuantitativo de PCR-tiempo real (qPCR), desarrollado localmente, utilizando la región no-estructural-5 (NS5), versus dos regiones tradicionalmente empleadas para la detección del virus, la región cápside/pre-membrana (C-prM), y la región noncodificante-3' (3'NC). Se recolectaron 167 muestras de suero de pacientes en fase aguda de la enfermedad. Las tres regiones génicas tuvieron perfiles de amplificación/coeficientes de correlación similares (0,987-0,999). Sin embargo, la región NS5 tuvo la eficiencia de amplificación más elevada para los cuatro serotipos (98-100%). Durante el proceso de validación, 41,1% (67/167) de las muestras de suero resultaron positivas para DENV al menos por dos de las regiones genómicas empleadas. Los valores de concordancia entre las regiones NS5/C-prM y NS5/3'NC fueron de 56,7% y 97%, respectivamente. La concordancia fue débil entre las regiones NS5/C-prM (k= 0,109; CI 95%), sin embargo, fue moderada entre las regiones NS5/3'NC (k= 0,489; CI 95%). El ensayo de tipificación uniplex en formato NS5/TaqMan® mostró alta sensibilidad (100%) que el protocolo C-prM/SYBRGreen®-I (76%). La validación externa del ensayo mostró una alta sensibilidad (100%), especificidad (78%) y acuerdo alto entre los ensayos utilizados. De acuerdo a los resultados obtenidos, la región NS5 ofrece la mayor opción para la detección y serotipificación del DENV en muestras clínicas.
Descritores: /genética
ABATTOIRS' UNTRANSLATED REGIONS/genética
Proteínas do Capsídeo/genética
Vírus da Dengue/genética
Dengue/virologia
Genoma Viral
Reação em Cadeia da Polimerase em Tempo Real
RNA Viral/análise
Proteínas não Estruturais Virais/genética
-Anticorpos Antivirais/sangue
Vírus da Dengue/classificação
Vírus da Dengue/imunologia
Vírus da Dengue/isolamento & purificação
Dengue/sangue
Diagnóstico Precoce
Ensaio de Imunoadsorção Enzimática
Imunoglobulina M/sangue
Compostos Orgânicos
Reprodutibilidade dos Testes
Reação em Cadeia da Polimerase em Tempo Real/métodos
Sensibilidade e Especificidade
Sorotipagem
Taq Polimerase
Cultura de Vírus
Limites: Humanos
Tipo de Publ: Estudo Comparativo
Estudo de Avaliação
Research Support, Non-U.S. Gov't
Estudo de Validação
Responsável: VE1.1 - Biblioteca Humberto Garcia Arocha


  8 / 21 LILACS  
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Id: lil-705289
Autor: Shi, Mei; Zhou, Yaping; Cao, Limin; Ding, Cuijun; Ji, Yun; Jiang, Qinbo; Liu, Xiping; Li, Xiang; Hou, Xueling; Peng, Hongjun; Shi, Weifeng.
Título: Expression of enterovirus 71 capsid protein VP1 in Escherichia coli and its clinical application
Fonte: Braz. j. microbiol;44(4):1215-1222, Oct.-Dec. 2013. ilus, graf, tab.
Idioma: en.
Resumo: The VPl gene of enterovirus 71 (EV71) was synthesized, construct a recombinant plasmid pET15b/VP1 and expressed in E. coli BL21. The recombinant VP1 protein could specifically react with EV71-infected patient sera without the cross-reaction with serum antibodies of coxsackievirus A16 (CA16), A4, A5, B3 and B5 as well as echovirus 6. In acute and convalescent phases, IgM and IgG antibodies of 182 serum samples were detected by ELISA with recombinant VP1 protein as a coated antigen. The results showed that the sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of IgM antibodies in serum samples for the diagnosis of EV71 infection were 90.1, 98.4, 98.8 and 88.7%, respectively; similarly, those of IgG antibodies in serum samples were 82.4, 89.1, 91.5 and 78.1%, respectively. Five of 80 samples (6.25%) from CA16infected patients were detected positive by ELISA with recombinant VP1 protein in which indicated the cross reactions and 0 of 5 samples from patients infected with other enteroviruses including CA4, CA5, CB3, CB5 and echovirus 6. Therefore, the recombinant VP1 protein of EV7l may provide a theoretical reference for establishing an effective antibody screening of IgM for EV71-infected patients with clinically suspected hand, foot, and mouth disease (HFMD).
Descritores: Anticorpos Antivirais/sangue
Proteínas do Capsídeo
Enterovirus Humano A/imunologia
Doença de Mão, Pé e Boca/diagnóstico
-Clonagem Molecular
Proteínas do Capsídeo/genética
Proteínas do Capsídeo/imunologia
Enterovirus Humano A/genética
Ensaio de Imunoadsorção Enzimática/métodos
Escherichia coli/genética
Expressão Gênica
Imunoglobulina G/sangue
Imunoglobulina M/sangue
Valor Preditivo dos Testes
Proteínas Recombinantes
Proteínas Recombinantes/genética
Proteínas Recombinantes/imunologia
Sensibilidade e Especificidade
Testes Sorológicos/métodos
Limites: Pré-Escolar
Feminino
Humanos
Lactente
Masculino
Tipo de Publ: Estudo de Avaliação
Responsável: BR1.1 - BIREME


  9 / 21 LILACS  
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Id: lil-678279
Autor: Memorias do Instituto Oswaldo Cruz; Martinez-Alvarez, Laura; Pina-Vazquez, Carolina; Zarco, Wilbert; Padilla-Noriega, Luis.
Título: The shift from low to high non-structural protein 1 expression in rotavirus-infected MA-104 cells
Fonte: Mem. Inst. Oswaldo Cruz;108(4):421-428, jun. 2013. graf.
Idioma: en.
Projeto: CONACyT; . CONACyT; . UNAM; . UNAM.
Resumo: A hallmark of group/species A rotavirus (RVA) replication in MA-104 cells is the logarithmic increase in viral mRNAs that occurs four-12 h post-infection. Viral protein synthesis typically lags closely behind mRNA synthesis but continues after mRNA levels plateau. However, RVA non-structural protein 1 (NSP1) is present at very low levels throughout viral replication despite showing robust protein synthesis. NSP1 has the contrasting properties of being susceptible to proteasomal degradation, but being stabilised against proteasomal degradation by viral proteins and/or viral mRNAs. We aimed to determine the kinetics of the accumulation and intracellular distribution of NSP1 in MA-104 cells infected with rhesus rotavirus (RRV). NSP1 preferentially localises to the perinuclear region of the cytoplasm of infected cells, forming abundant granules that are heterogeneous in size. Late in infection, large NSP1 granules predominate, coincident with a shift from low to high NSP1 expression levels. Our results indicate that rotavirus NSP1 is a late viral protein in MA-104 cells infected with RRV, presumably as a result of altered protein turnover.
Descritores: Proteínas do Capsídeo/metabolismo
Regulação Viral da Expressão Gênica
Rotavirus/metabolismo
Proteínas não Estruturais Virais/metabolismo
-Linhagem Celular
RNA Viral/genética
Rotavirus/fisiologia
Replicação Viral
Limites: Animais
Cobaias
Tipo de Publ: Research Support, Non-U.S. Gov't
Responsável: BR1.1 - BIREME


  10 / 21 LILACS  
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Id: lil-664764
Autor: Rezende, Joffre M. de.
Título: Envelope viral / Viral envelope
Fonte: Rev. patol. trop;41(3):373-374, jul.-set. 2012.
Idioma: pt.
Descritores: Proteínas do Capsídeo
Responsável: BR15.1 - Biblioteca de Ciências Biomédicas



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