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Id: lil-426908
Autor: Carnieli Junior, Pedro; Ventura, Armando Moraes; Durigon, Edison Luiz.
Título: Digoxigenin-labeled probe for rabies virus nucleoprotein gene detection
Fonte: Rev. Soc. Bras. Med. Trop;39(2):159-162, mar.-abr. 2006. ilus, tab.
Idioma: en.
Resumo: A digoxigenin-labeled probe was produced from the Pasteur virus strain for the detection of the rabies virus N gene. The probe hybridization was performed from amplified N gene obtained by reverse transcription polymerase chain reaction and the results by RT-PCR and hybridization showed 100 percent agreement. The hybridization, when carried out in products amplified by RT-PCR, increases the sensitivity of this technique even more and confers specificity to the diagnosis. The technique described in this work will be useful in rabies diagnosis laboratories, once the cost is compatible with traditional rabies diagnostic techniques.
Descritores: DNA Viral/genética
Digoxigenina
Nucleoproteínas/genética
Vírus da Raiva/genética
Raiva/diagnóstico
-Medições Luminescentes
Southern Blotting
Quirópteros
Sondas de DNA
Cavalos
Hibridização In Situ
Raiva/virologia
Reação em Cadeia da Polimerase Via Transcriptase Reversa
Sensibilidade e Especificidade
Ovinos
Limites: Humanos
Animais
Bovinos
Cães
Responsável: BR1.1 - BIREME


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Id: lil-426796
Autor: Avila-Campos, Mario J; Rivera, Irma N; Nakano, Viviane.
Título: Genetic diversity of oral Fusobacterium nucleatum isolated from patients with different clinical conditions
Fonte: Rev. Inst. Med. Trop. Säo Paulo;48(2):59-63, Mar,-Apr. 2006. ilus, tab.
Idioma: en.
Projeto: Fundação de Amparo à Pesquisa do Estado de São Paulo.
Resumo: Neste estudo foi avaliada a diversidade genética de 23 amostras de Fusobacterium nucleatum isoladas da cavidade bucal de 15 pacientes com doença periodontal, de oito cepas isoladas de sete indivíduos sadios, de nove isoladas de nove pacientes com AIDS e de duas isoladas de dois macacos Cebus apella. Pela ação da enzima EcoRI sobre o DNA bacteriano foram reconhecidos 28 ribotipos agrupados de A a J. Os isolados testados formaram 24 ribotipos os quais foram contidos nos grupos A, B, C, D, E e F, e as três cepas de referência e dois isolados clínicos de A. actinomycetemcomitans e E. coli CDC formaram quatro diferentes ribotipos contidos nos grupos G, H, I e J. Em adição, as nove cepas de F. nucleatum isoladas de pacientes com AIDS, seis pertenciam ao grupo C e três ao grupo D. Usando-se a ribotipagem foi possível distinguir F. nucleatum isolados de diferentes origens.
Descritores: Variação Genética
DNA Bacteriano/análise
Infecções por Fusobacterium/microbiologia
Fusobacterium nucleatum/genética
Doenças Periodontais/microbiologia
-Infecções Oportunistas Relacionadas com a AIDS/microbiologia
Técnicas de Tipagem Bacteriana
Southern Blotting
Cebus/microbiologia
Ribotipagem
Limites: Humanos
Animais
Adolescente
Adulto
Tipo de Publ: Research Support, Non-U.S. Gov't
Responsável: BR1.1 - BIREME


  3 / 81 LILACS  
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Id: biblio-1022493
Autor: Liu, Dongmei; Zhu, Hanyu; Chen, Yue; Zheng, Liesheng; Chen, Liguo; Ma, Aimin.
Título: Cloning and heterologous expression of a hydrophobin gene Ltr. hyd from the tiger milk mushroom Lentinus tuber-regium in yeast-like cells of Tremella fuciformis
Fonte: Electron. j. biotechnol;32:6-12, Mar. 2018. tab, graf, ilus.
Idioma: en.
Projeto: National Natural Science Foundation of China (NSFC).
Resumo: Background: Hydrophobins are small proteins secreted by filamentous fungi, which show a highly surface activity. Because of the signally self-assembling abilities and surface activities, hydrophobins were considered as candidates in many aspects, for example, stabilizing foams and emulsions in food products. Lentinus tuber-regium, known as tiger milk mushroom, is both an edible and medicinal sclerotium-producing mushroom. Up to now, the hydrophobins of L. tuber-regium have not been identified. Results: In this paper, a Class I hydrophobin gene, Ltr.hyd, was cloned from L. tuber-regium and expressed in the yeast-like cells of Tremella fuciformis mediated by Agrobacterium tumefaciens. The expression vector pGEH-GH was under the control of T. fuciformis glyceraldehyde-3-phosphate dehydrogenase gene (gpd) promoter. The integration of Ltr.hyd into the genome of T. fuciformis was confirmed by PCR, Southern blot, fluorescence observation and quantitative real-time PCR (qRT-PCR). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated that recombinant hydrophobin rLtr.HYD with an expected molecular mass of 13 kDa was extracted. The yield of rLtr.HYD was 0.66 mg/g dry weight. The emulsifying activity of rLtr.HYD was better than the typical food emulsifiers sodium caseinate and Tween 20. Conclusions: We evaluated the emulsifying property of hydrophobin Ltr.HYD, which can be potentially used as a food emulsifier.
Descritores: Basidiomycota/metabolismo
Proteínas Fúngicas/genética
Lentinula/genética
Lentinula/metabolismo
-Transformação Genética
Basidiomycota/enzimologia
Leveduras
Proteínas Fúngicas/metabolismo
Southern Blotting
Clonagem Molecular
Agrobacterium tumefaciens/metabolismo
Análise de Sequência
Emulsificantes
Eletroforese em Gel de Poliacrilamida
Reação em Cadeia da Polimerase em Tempo Real
Gliceraldeído-3-Fosfato Desidrogenases/metabolismo
Microscopia de Fluorescência
Responsável: CL1.1 - Biblioteca Central


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Id: biblio-1008414
Autor: Liu, Juhua; Gao, Pengzhao; Sun, Xiuxiu; Zhang, Jing; Sun, Peiguang; Wang, Jiashui; Jia, Caihong; Zhang, Jianbin; Hu, Wei; Xu, Biyu; Jin, Zhiqiang.
Título: Efficient regeneration and genetic transformation platform applicable to five Musa varieties
Fonte: Electron. j. biotechnol;25:33-38, ene. 2017. tab, ilus.
Idioma: en.
Projeto: Modern Agro-industry Technology Research System; . National Nonprofit Institute Research Grant of Institute of Tropical Bioscience and Biotechnology CATAS-ITBB.
Resumo: Background: Banana (Musa spp.) is an important staple food, economic crop, and nutritional fruit worldwide. Conventional breeding has been seriously hampered by their long generation time, polyploidy, and sterility of most cultivated varieties. Establishment of an efficient regeneration and transformation system for banana is critical to its genetic improvement and functional genomics. Results: In this study, a vigorous and repeatable transformation system for banana using direct organogenesis was developed. The greatest number of shoots per explant for all five Musa varieties was obtained using Murashige and Skoog medium supplemented with 8.9 µM benzylaminopurine and 9.1 µM thidiazuron. One immature male flower could regenerate 380­456, 310­372, 200­240, 130­156, and 100­130 well-developed shoots in only 240­270 d for Gongjiao, Red banana, Rose banana, Baxi, and Xinglongnaijiao, respectively. Longitudinal sections of buds were transformed through particle bombardment combined with Agrobacterium-mediated transformation using a promoterless ß-glucuronidase (GUS) reporter gene; the highest transformation efficiency was 9.81% in regenerated Gongjiao plantlets in an optimized selection medium. Transgenic plants were confirmed by a histochemical assay of GUS, polymerase chain reaction, and Southern blot. Conclusions: Our robust transformation platform successfully generated hundreds of transgenic plants. Such a platform will facilitate molecular breeding and functional genomics of banana.
Descritores: Musa/crescimento & desenvolvimento
Musa/genética
-Regeneração
Transformação Genética
Imuno-Histoquímica
Southern Blotting
Reação em Cadeia da Polimerase
Plantas Geneticamente Modificadas
Agrobacterium tumefaciens/fisiologia
Musa/microbiologia
Organogênese Vegetal
Glucuronidase
Responsável: CL1.1 - Biblioteca Central


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Zatz, Mayana
Bueno, Maria Rita Passos
Machado, Luís dos Ramos
Yacubian, Elza Márcia Targas
Id: lil-165394
Autor: Zatz, Mayana; Bueno, Maria Rita Passos.
Título: Testes moleculares em medicina: quais e quando / Molecular tests in medicine: what and when
Fonte: In: Nitrini, Ricardo; Machado, Luís dos Ramos; Yacubian, Elza Marcia Targas; Rabello, Getúlio Daré. Condutas em neurologia: 1995. Säo Paulo, Clínica Neurológica HC/FMUSP, 1995. p.69-69.
Idioma: pt.
Descritores: Aberrações Cromossômicas/diagnóstico
Genes/fisiologia
Biologia Molecular
-Projeto Genoma Humano
Southern Blotting
Reação em Cadeia da Polimerase
Mapeamento Cromossômico
Limites: Humanos
Responsável: BR1.1 - BIREME
BR1.1/2673.06; BR73.1; WL100, N731c. 1968


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Id: lil-751586
Autor: Oliveira, Guilherme Marx de.
Título: Uso de técnicas moleculares na detecção de DNA parasitário e no estudo da variabilidade genética em diferentes fragmentos teciduais de cães naturalmente infectados por Leishmania (Viannia) braziliensis / Use of molecular techniques in parasite DNA detection and genetic variability study in different tissue fragments from naturally infected dogs by Leishmania (Viannia) braziliensis.
Fonte: Rio de Janeiro; s.n; 2013. 50 p. ilus, tab, graf.
Idioma: pt.
Tese: Apresentada a Instituto de Pesquisa Clínica Evandro Chagas para obtenção do grau de Mestre.
Resumo: A leishmaniose tegumentar americana (LTA), em cães, costuma estar associada à resposta humoral baixa ou mesmo negativa, o que inviabiliza os métodos sorológicos convencionais, como ferramenta única no diagnóstico. [...] Métodos convencionais de diagnóstico parasitológico, não têm sido capazes de detectar a presença do parasito em outros sítios anatômicos diferentes da lesão cutânea, em cães naturalmente infectados por Leishmania (Viannia) braziliensis. Diante dos questionamentos sobre o papel do cão doméstico no ciclo de transmissão da LTA e sobre o valor de métodos diagnósticos aplicados na rotina, principalmente em áreas de sobreposição da forma tegumentar e visceral, faz-se necessário a avaliação desses animais sob diversos aspectos. O presente estudo teve como objetivo empregar a PCR específica associada à hibridização molecular e a técnica da reação em cadeia da polimerase com primer único em condições de baixa estringência (LSSP-PCR) visando detectar a presença de DNA parasitário e investigar a variabilidade genética de populações parasitárias presentes em diferentes tecidos de cães naturalmente infectados por L. (V.) braziliensis. Os animais foram selecionados entre os cães sororeatores para Leishmania procedentes de cidades do estado do Rio de Janeiro e encaminhados para eutanásia ao Laboratório de Pesquisa Clínica em Dermatozoonoses em Animais Domésticos do Instituto de Pesquisa Clínica Evandro Chagas da Fundação Oswaldo Cruz. Durante a necropsia, foram obtidas amostras de lesão cutânea, pele íntegra (região escapular e abdominal), baço, fígado e linfonodos (poplíteo e cervical)...

In dogs, American tegumentary leishmaniasis (ATL) is usually associated to a low humoral responseor even to negative results which turns not unfeasible the conventional serological methods. [...] Conventional methods ofparasitological diagnosis have failed to detect the presence of the parasite anatomic sites others thanthe cutaneous lesion, in dogs naturally infected by Leishmania (Viannia) braziliensis. Regarding thequestions on the rule of domestic dogs in the transmission cycle of ATL and, on the applicability ofdiagnostic methods mainly in areas where both visceral and tegumentary disease are found, theevaluation of these animals under different aspects is needed. The objective of this project is to applyspecific PCR assays associated to molecular hybridization and the technique of Low-StringencySpecific-Single Primer – PCR (LSSP-PCR) in order to detect the presence of parasite DNA andevaluate the genetic variability of parasite populations found in different tissues of dogs naturallyinfected by Leishmania (V.) braziliensis...
Descritores: Leishmania braziliensis
Leishmaniose Cutânea/diagnóstico
Leishmaniose Cutânea/epidemiologia
Leishmaniose Cutânea/transmissão
-Southern Blotting
Reação em Cadeia da Polimerase
Limites: Cães
Responsável: BR15.1 - Biblioteca de Ciências Biomédicas


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Id: lil-723999
Autor: Sun, Mingyue; Wang, Yue; Chen, Zhongju; Zhu, Xuhui; Tian, Lei; Sun, Ziyong.
Título: The first report of the vanC1 gene in Enterococcus faecium isolated from a human clinical specimen
Fonte: Mem. Inst. Oswaldo Cruz;109(6):712-715, 09/09/2014. graf.
Idioma: en.
Projeto: China National Mega Project.
Resumo: The vanC1 gene, which is chromosomally located, confers resistance to vancomycin and serves as a species marker for Enterococcus gallinarum. Enterococcus faecium TJ4031 was isolated from a blood culture and harbours the vanC1gene. Polymerase chain reaction (PCR) assays were performed to detect vanXYc and vanTc genes. Only the vanXYc gene was found in the E. faecium TJ4031 isolate. The minimum inhibitory concentrations of vancomycin and teicoplanin were 2 µg/mL and 1 µg/mL, respectively. Real-time reverse transcription-PCR results revealed that the vanC1and vanXYc genes were not expressed. Pulsed-field gel electrophoresis and southern hybridisation results showed that the vanC1 gene was encoded in the chromosome. E. faecalis isolated from animals has been reported to harbour vanC1gene. However, this study is the first to report the presence of the vanC1gene in E. faecium of human origin. Additionally, our research showed the vanC1gene cannot serve as a species-specific gene of E. gallinarum and that it is able to be transferred between bacteria. Although the resistance marker is not expressed in the strain, our results showed that E. faecium could acquire the vanC1gene from different species.
Descritores: Proteínas de Bactérias/genética
Enterococcus faecium/genética
Genes Bacterianos/genética
Enterococos Resistentes à Vancomicina/genética
-Antibacterianos/farmacologia
Southern Blotting
Proteínas de Bactérias/sangue
Eletroforese em Gel de Campo Pulsado
Enterococcus faecalis/genética
Enterococcus faecium/efeitos dos fármacos
Enterococcus/efeitos dos fármacos
Enterococcus/genética
Hibridização In Situ/métodos
Testes de Sensibilidade Microbiana
Tipagem de Sequências Multilocus
Família Multigênica/fisiologia
Reação em Cadeia da Polimerase
Teicoplanina/farmacologia
Resistência a Vancomicina/genética
Vancomicina/farmacologia
Limites: Humanos
Tipo de Publ: Research Support, Non-U.S. Gov't
Responsável: BR1.1 - BIREME


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Id: lil-640518
Autor: Zhang, Wei-Wei; Yang, Ming-Ming; Li, Heng-xin; Wang, Dun.
Título: Construction of recombinant Bacillus subtilis strains for efficient pimelic acid synthesis
Fonte: Electron. j. biotechnol;14(6):1-1, Nov. 2011. ilus, tab.
Idioma: en.
Projeto: Science and Technology Ministry. National New Productions Project.
Resumo: As a precursor, pimelic acid plays an important role in biotin biosynthesis pathway of Bacillus subtilis. Fermentations supplemented with pimelic acid could improve the production of biotin, however, with a disadvantage-high cost. So it is necessary to improve the biosynthesis of pimelic acid via genetic engineering in B. subtilis. In this study, we constructed a recombinant B. subtilis strain for improving the synthesis of pimelic acid, in which a maltose-inducible Pglv promoter was inserted into the upstream of the cistron bioI-orf2-orf3 and, meanwhile, flanked by the tandem cistrons via a single crossover event. The copy number of the integrant was amplified by high-concentration resistance screen and increased to 4-5 copies. The production of pimelic acid from multiple copies integrant was about 4 times higher than that from single copy (1017.13 ug/ml VS. 198.89 μg/ml). And when induced by maltose the production of pimelic acid was about 2 times of that under non-induction conditions (2360.73 μg/ml VS. 991.59 ug/ml). Thus, these results demonstrated that the production of pimelic acid was improved obviously through reconstructed B. subtilis. It also suggested that our expression system provided a convenient source of pimelic acid that would potentially lower the cost of production of biotin from engineered B. subtilis.
Descritores: Ácidos Pimélicos/metabolismo
Bacillus subtilis
-Southern Blotting
Cromatografia Líquida de Alta Pressão
Regiões Promotoras Genéticas
Recombinação Genética
Responsável: CL1.1 - Biblioteca Central


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Id: lil-639657
Autor: Cánepa, Mariana; Deninghoff, Valeria; Perazzo, Florencia; Paesani, Fernando; Nieto, Silvana; García, Alejandro; Avagnina, Alejandra; Elsner, Boris.
Título: Amplificación del oncogén Her-2/neu en el carcinoma mamario
Fonte: Medicina (B.Aires);72(1):88-89, feb. 2012. tab.
Idioma: es.
Descritores: Neoplasias da Mama/genética
Amplificação de Genes
/genética
GENES, ERBB-TEMEFOS/genética
-Southern Blotting
Western Blotting
Hibridização In Situ/métodos
Limites: Feminino
Humanos
Tipo de Publ: Carta
Responsável: AR1.2 - Instituto de Investigaciónes Epidemiológicas


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Id: lil-626737
Autor: Liu, Feng; Zhao, Yi-Ying; Zheng, Jin-Gui.
Título: High-level expression of modified gene encoding human adiponectin in transgenic rice
Fonte: Biol. Res;44(4):369-375, 2011. ilus, tab.
Idioma: en.
Projeto: research program of Fujian, China; . Doctor Funds of Xinjiang Bingtuan.
Resumo: Adiponectin is a polypeptide specifically secreted from human adipocytes, and its deficiency is closely linked to increased obesity and type II diabetes. There is an urgent demand for large-scale production of human adiponectin for pharmaceutical applications. Here, we report that we have successfully obtained a high-level of expression of modified genes encoding human adiponectin in transgenic rice. The 735 bp cDNA of the native human sequence was adopted to rice codon usage, fused to the translation initiation sequence in the N terminus and to the KDEL signal sequence in the C terminus. An amplification promoting sequence acting as an enhancer of transcription was also introduced to enhance gene expression. The presence of the transgene and mRNA transcripts was confirmed by PCR, Southern blot and RT-PCR. Western blot analysis revealed that a protein of approximately 30 kDa was produced in rice leaves. ELISA analysis was used to determine the amount of recombinant adiponectin in transformants with the modified gene in up to 0.32% of total soluble leaf protein. Our results establish the feasibility of high-level expression of recombinant human adiponectin in transgenic rice.
Descritores: Adiponectina/genética
Oryza/genética
Plantas Geneticamente Modificadas/genética
-Adiponectina/metabolismo
Southern Blotting
Códon
DNA Complementar
Regulação da Expressão Gênica de Plantas
Oryza/química
Reação em Cadeia da Polimerase Via Transcriptase Reversa
Tipo de Publ: Research Support, Non-U.S. Gov't
Responsável: CL1.1 - Biblioteca Central



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