Base de dados : LILACS
Pesquisa : D08.811.150.280.260.240 [Categoria DeCS]
Referências encontradas : 4 [refinar]
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Id: biblio-1021557
Autor: Li, Hedan; Hao, Chengwei; Xu, Daqing.
Título: Development of a novel vector for cloning and expressing extremely toxic genes in Escherichia coli
Fonte: Electron. j. biotechnol;30:88-94, nov. 2017. tab, ilus, graf.
Idioma: en.
Projeto: National Natural Science Foundation of China; . Specialized Research Fund for the Doctoral Programme of Higher Education of China.
Resumo: Background: Escherichia coli has been widely used as a host to clone and express heterologous genes. However, there are few vectors available for cloning and expressing extremely toxic genes, which limits further basic and applied research on extremely toxic proteins. Results: In this study, a novel vector pAU10 was constructed in E. coli. pAU10 utilizes the combination of the efficient but highly repressible T7-lacO promoter/operator and the strong rrnBT2 transcriptional terminator upstream of the T7 promoter to strictly control unwanted transcription of the extremely toxic gene; in addition, the trp promoter/operator is oriented opposite to the T7 promoter to control the production of the antisense RNA that may block the translation of leaky mRNA. Without the supplementation of IPTG and L-tryptophan in the culture medium, transcription of the extremely toxic gene by the T7 promoter is highly repressed, and the trp promoter produces the antisense RNA, which strictly prevents unwanted expression of the extremely toxic protein in E. coli. With the supplementation of IPTG and L-tryptophan, the T7 promoter efficiently transcribes the extremely toxic gene, and the trp promoter does not produce the antisense RNA, ensuring efficient expression of the extremely toxic protein in E. coli. Tight regulation and efficiency of expression of an extremely toxic gene cloned in the vector pAU10 were confirmed by cloning and expressing the restriction endonuclease-encoding gene bamHI without its corresponding methylase gene in E. coli JM109(DE3). Conclusion: pAU10 is a good vector used for cloning and expressing extremely toxic genes in E. coli.
Descritores: Proteínas de Escherichia coli/toxicidade
Escherichia coli/genética
Vetores Genéticos
-Triptofano/metabolismo
Desoxirribonuclease BamHI/metabolismo
Western Blotting
Reação em Cadeia da Polimerase
RNA Antissenso
Regiões Promotoras Genéticas
Clonagem Molecular
Eletroforese em Gel de Poliacrilamida
Proteínas Correpressoras
Genes Bacterianos
Isopropiltiogalactosídeo/metabolismo
Responsável: CL1.1 - Biblioteca Central


  2 / 4 LILACS  
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Id: lil-321732
Autor: Figueiredo, M. S.
Título: Location and rapid analysis of the intragenic BamHI polymorphic site of the factor IX gene
Fonte: Braz. j. med. biol. res = Rev. bras. pesqui. méd. biol;27(5):1117-1121, May 1994.
Idioma: en.
Resumo: We report the precise location and a polymerase chain reaction (PCR)-based method for the analysis of the intragenic BamHI polymorphism of the factor IX (FIX) gene. After screening DNA samples from the Brazilian Black population by a selective amplification of a segment of the FIX gene containing entire exon 2, intron 2, and exon 3 followed by digestion of PCR products with BamHI, we were able to identify individuals presenting the polymorphic BamHI site. By DNA sequencing of selected samples, the dimorphic base was located at nucleotide number 6575 (intron 2). The PCR method outlined here allows rapid and easy analysis of this polymorphism. Its application may be particularly useful for carrier detection and prenatal diagnosis of hemophilia B in the Black population, thus far the only one that has been shown to be polymorphic for this site. Because of its apparent restrictive pattern it may also be used in combination with other markers for estimates of racial admixture in mixed populations in which the Black population is present (as is the case for most countries in the American continent).
Descritores: Desoxirribonuclease BamHI
Fator IX
Polimorfismo Genético/genética
-Grupo com Ancestrais do Continente Africano
Sequência de Bases
Dados de Sequência Molecular
Reação em Cadeia da Polimerase/métodos
Análise de Sequência de DNA
Limites: Feminino
Seres Humanos
Masculino
Tipo de Publ: Research Support, Non-U.S. Gov't
Responsável: BR1.1 - BIREME


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Id: lil-273441
Autor: Pumariega, Tania; Savón, Clara; Muné, Mayra; Cancio, Reynel; González, Grehete; Valdivia, Angel; González, Zoila; Goyenechea, Angel.
Título: Isolation and identification of Adenovirus in hospitalized children, under five years, with acute respiratory disease, in Havana, Cuba
Fonte: Mem. Inst. Oswaldo Cruz;95(6):859-61, Nov.-Dec. 2000. ilus, tab.
Idioma: en.
Resumo: Nine Adenovirus (Ad) strains isolated in Cuba, from 128 nasopharingeal swab specimens of children below five years old, with acute respiratory diseases, during 1996 and 1997, were studied by restriction enzyme analysis of genomic DNA with two endonucleases BamH I and Sma I. All different fragment patterns were compared with the respective prototypes. The identified adenoviruses were Ad 1 (n=4), Ad 2 (n=1) and Ad 6 (n=4). Males were more frequently infected than females. The analysis of the occurrence of these Adenovirus strains of subgenus C revealed that Ad 1 and Ad 6 were the predominant serotypes in 1996 and in 1997, respectively
Descritores: Adenovírus Humanos/isolamento & purificação
Doenças Respiratórias/virologia
-Doença Aguda
Infecções por Adenovirus Humanos/diagnóstico
Cuba
Desoxirribonuclease BamHI
Enzimas de Restrição do DNA
Limites: Seres Humanos
Masculino
Feminino
Lactente
Pré-Escolar
Responsável: BR1.1 - BIREME


  4 / 4 LILACS  
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Id: lil-210964
Autor: Galosi, C. M; Norimine, J; Echeverría, M. G; Oliva, G. A; Nosetto, E. O; Etcheverrigaray, M. E; Tohya, Y; Mikami, T.
Título: Diversity of genomic electropherotypes of naturally occurring equine herpesvirus 1 isolates in Argentina
Fonte: Braz. j. med. biol. res = Rev. bras. pesqui. méd. biol;31(6):771-4, jun. 1998. ilus.
Idioma: en.
Resumo: The genomes of 10 equine herpesvirus 1 (EHV-1) strains isolated in Argentina from 1979 to 1991, and a Japanese HH1 reference strain were compared by restriction endonuclease analysis. Two restriction enzymes, Bam HI and Bgl II, were used and analysis of the electropherotypes did not show significant differences among isolates obtained from horses with different clinical signs. This suggests that the EHV-1 isolates studied, which circulated in Argentina for more than 10 years, belong to a single genotype
Descritores: Variação Genética
Genoma
Herpesvirus Equídeo 1/genética
-Argentina
Desoxirribonuclease BamHI
Desoxirribonucleases de Sítio Específico do Tipo II
Eletroforese
Herpesvirus Equídeo 1/isolamento & purificação
Tipo de Publ: Revisão
Estudo Comparativo
Research Support, Non-U.S. Gov't
Responsável: BR1.1 - BIREME



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