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Registro Completo |
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Biblioteca(s): |
Embrapa Gado de Corte. |
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Data corrente: |
22/03/2011 |
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Data da última atualização: |
09/09/2014 |
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Tipo da produção científica: |
Artigo em Anais de Congresso |
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Autoria: |
ANDREOTTI, R.; SOARES, M.; CUNHA, R. C.; LEITE, F. P. L. |
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Afiliação: |
RENATO ANDREOTTI E SILVA, CNPGC; UFPEL; UFPEL; UFPEL. |
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Título: |
Heterologous expression in Pichia pastoris of gene BMTI optimized. |
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Ano de publicação: |
2010 |
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Fonte/Imprenta: |
In: INTERNATIONAL CONGRESS OF PARASITOLOGY, 12., 2010, Melbourne. Conference abstracts. Melbourne: The Australian Society for Parasitology:World Federation of Parasitologists, 2010. |
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Páginas: |
p.149-152 |
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Idioma: |
Inglês
Português |
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Notas: |
ICOPA XII |
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Conteúdo: |
The cattle tick, Rhipicephalus microplus, is the main tick species that undermines productivity losses in livestock resulting in the production of milk and meat throughout Brazil. Its control is primarily chemical and faces problems due to the resistance of ticks to several active principles used. The immune control is a viable alternative to tick control, and the protein BMTI is a strong candidate as a vaccine antigen. The aim of this work was to synthesize the sequencing of BMTI (BmTIS), optimizing it to "codon usage" of Pichia pastoris in an attempt to improve its expression. This sequence was cloned in eukaryotic expression vector pPICZaA, respecting the make reading the AOX1 promoter, present in the plasmid and that is strongly induced by the presence of methanol, following the sign "a-factor for protein secretion, and the tail of poly-histidine. The resulting plasmid, pPICZaBmTIS, X33 was transformed into strains of P. pastoris. Clones were characterized by PCR using primers of the vector and also by dot blot analysis using anti-histidine. A clone (X33BmTIS17) was sequenced and selected to be induced. A single colony clone X33BmTIS17 was inoculated with 25mL of minimal medium with glycerol and incubated at 200rpm, 300C, overnight. The culture was centrifuged at 1500 g, 5min and resuspended in minimal medium with methanol. The supernatant was subjected to SDS-PAGE 0.7%, which showed a major band with the approximate size of 49 kDa. In Western bloting, a band at the same time was recognized by the anti-histidine and by bovine serum immunized with a synthetic based pepitídeo (BmTI). Thus, a recombinant protein (rBmTI) was
expressed, and was recognized by the anti-histidine and characterized as potential antigenic by bovine serum immunized with the synthetic pepitídeo. MenosThe cattle tick, Rhipicephalus microplus, is the main tick species that undermines productivity losses in livestock resulting in the production of milk and meat throughout Brazil. Its control is primarily chemical and faces problems due to the resistance of ticks to several active principles used. The immune control is a viable alternative to tick control, and the protein BMTI is a strong candidate as a vaccine antigen. The aim of this work was to synthesize the sequencing of BMTI (BmTIS), optimizing it to "codon usage" of Pichia pastoris in an attempt to improve its expression. This sequence was cloned in eukaryotic expression vector pPICZaA, respecting the make reading the AOX1 promoter, present in the plasmid and that is strongly induced by the presence of methanol, following the sign "a-factor for protein secretion, and the tail of poly-histidine. The resulting plasmid, pPICZaBmTIS, X33 was transformed into strains of P. pastoris. Clones were characterized by PCR using primers of the vector and also by dot blot analysis using anti-histidine. A clone (X33BmTIS17) was sequenced and selected to be induced. A single colony clone X33BmTIS17 was inoculated with 25mL of minimal medium with glycerol and incubated at 200rpm, 300C, overnight. The culture was centrifuged at 1500 g, 5min and resuspended in minimal medium with methanol. The supernatant was subjected to SDS-PAGE 0.7%, which showed a major band with the approximate size of 49 kDa. In Western bloting, a band at the same... Mostrar Tudo |
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Thesagro: |
Carrapato; Sanidade Animal. |
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Thesaurus Nal: |
Rhipicephalus microplus. |
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Categoria do assunto: |
H Saúde e Patologia
L Ciência Animal e Produtos de Origem Animal |
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Marc: |
LEADER 02512naa a2200217 a 4500
001 1883447
005 2014-09-09
008 2010 bl --- 0-- u #d
100 1 $aANDREOTTI, R.
245 $aHeterologous expression in Pichia pastoris of gene BMTI optimized.
260 $c2010
300 $ap.149-152
500 $aICOPA XII
520 $aThe cattle tick, Rhipicephalus microplus, is the main tick species that undermines productivity losses in livestock resulting in the production of milk and meat throughout Brazil. Its control is primarily chemical and faces problems due to the resistance of ticks to several active principles used. The immune control is a viable alternative to tick control, and the protein BMTI is a strong candidate as a vaccine antigen. The aim of this work was to synthesize the sequencing of BMTI (BmTIS), optimizing it to "codon usage" of Pichia pastoris in an attempt to improve its expression. This sequence was cloned in eukaryotic expression vector pPICZaA, respecting the make reading the AOX1 promoter, present in the plasmid and that is strongly induced by the presence of methanol, following the sign "a-factor for protein secretion, and the tail of poly-histidine. The resulting plasmid, pPICZaBmTIS, X33 was transformed into strains of P. pastoris. Clones were characterized by PCR using primers of the vector and also by dot blot analysis using anti-histidine. A clone (X33BmTIS17) was sequenced and selected to be induced. A single colony clone X33BmTIS17 was inoculated with 25mL of minimal medium with glycerol and incubated at 200rpm, 300C, overnight. The culture was centrifuged at 1500 g, 5min and resuspended in minimal medium with methanol. The supernatant was subjected to SDS-PAGE 0.7%, which showed a major band with the approximate size of 49 kDa. In Western bloting, a band at the same time was recognized by the anti-histidine and by bovine serum immunized with a synthetic based pepitídeo (BmTI). Thus, a recombinant protein (rBmTI) was expressed, and was recognized by the anti-histidine and characterized as potential antigenic by bovine serum immunized with the synthetic pepitídeo.
650 $aRhipicephalus microplus
650 $aCarrapato
650 $aSanidade Animal
700 1 $aSOARES, M.
700 1 $aCUNHA, R. C.
700 1 $aLEITE, F. P. L.
773 $tIn: INTERNATIONAL CONGRESS OF PARASITOLOGY, 12., 2010, Melbourne. Conference abstracts. Melbourne: The Australian Society for Parasitology:World Federation of Parasitologists, 2010.
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Registro original: |
Embrapa Gado de Corte (CNPGC) |
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| Registros recuperados : 55 | |
| 4. |  | SANTOS JUNIOR, A. G.; CUNHA, R. C.; ANDREOTTI, R.; LEITE, F. P. L. Avaliação de diferentes métodos de purificação da proteína bm86-cg expressa em pichia pastoris. In: CONGRESSO BRASILEIRO DE PARASITOLOGIA VETERINÁRIA, 16., 2010, Campo Grande, MS. [Anais...]. Campo Grande, MS: Colégio Brasileiro de Parasitologia Veterinária, 2010. 1 CD-ROM. 1 p.| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Gado de Corte. |
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| 6. | | ANDREOTTI, R.; SAIMO-KAHWA, M.; CUNHA, R. C.; BARROS, J. C.; LEITE, F. P. L. Characterization and Production of recombinant protein Ra92A from Rhipicephalus appendiculatus aiming tick vaccine antigen in Uganda. In: INTERNATIONAL CONGRESS OF PARASITOLOGY, 13., 2014, Mexico. Conference abstracts... Mexico City, Mexico: Academic Committee ICOPA Congress, 2014.| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Gado de Corte. |
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| 8. | | CUNHA, R. C.; LEITE, F. P. L.; PINTO, L. da S.; SATO, T.; ANDREOTTI, R. Subclonagem do gene Bm86-CG em pPIC9 para expressão heteróloga em Pichia pastoris. In: CONGRESSO BRASILEIRO DE PARASITOLOGIA 21.; ENCONTRO DE PARASITOLOGIA DO MERCOSUL, 2., 2009, Foz do Iguaçu. Novos horizontes em parasitologia: anais. Foz do Iguaçu: SBP, 2009. p. 1298. Revista Patologia Tropical, v. 38, jul.-set 2009. Suplemento 2.| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Gado de Corte. |
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| 12. | | SOUZA, C. C.; XAVIER, A.; LEITE, F. P.; SANTANA, R. C.; PAIVA, A. N. de. Densidade de minicepas em minijardim clonal na produção de mudas de eucalipto. Pesquisa Florestal Brasileira, Colombo, v. 34, n. 77, p. 49-56, jan./mar. 2014.| Biblioteca(s): Embrapa Florestas. |
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| 14. | | LEITE, F. P.; BARROS, N. F. de; SANS, L. M. A; FABRES, A. S. Regime hídrico do solo sob povoamento de eucalipto, floresta nativa e pastagem, na região de Guanhães-MG. Revista Árvore, Viçosa, v. 21, n. 4, p. 455-462, 1997.| Tipo: Artigo em Periódico Indexado |
| Biblioteca(s): Embrapa Milho e Sorgo. |
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| 16. | | PIRES, L. S.; SILVA, M. L. N.; CURI, N.; LEITE, F. P.; BRITO, L. de F. Erosão hídrica pós-plantio em florestas de eucalipto na região centro-leste de Minas Gerais. Pesquisa Agropecuária Brasileira, Brasília, DF, v. 41, n. 4, p. 687-695, abr. 2006 Título em inglês: Water erosion in post-planting eucalyptus forests at center-east region of Minas Gerais State, Brazil.| Biblioteca(s): Embrapa Unidades Centrais. |
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| 18. | | SILVA, M. A. da; SILVA, M. L. N.; CURI, N.; AVANZI, J. C.; LEITE, F. P. Sistemas de manejo em plantios florestais de eucalipto e perdas de solo e água na região do Vale do Rio Roce, MG. Ciência Florestal, Santa Maria, v. 21, n. 4, p. 765-776, out./dez. 2011.| Tipo: Artigo em Periódico Indexado | Circulação/Nível: B - 1 |
| Biblioteca(s): Embrapa Pesca e Aquicultura. |
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| 19. | | SILVA, A. R.; DIAS JUNIOR, M. de S.; LEITE, F. P.; CARVALHO, E. J. M.; VELOSO, C. A. C. Avaliação do efeito da camada de resíduos florestais na pressão de preconsolidação da um latossolo amarelo. In: CONGRESSO BRASILEIRO DE CIÊNCIA DO SOLO, 33., 2011, Uberlândia. Solos nos biomas brasileiros: sustentabilidade e mudanças climáticas: anais. [Uberlândia]: SBCS: UFU, ICIAG, 2011.| Tipo: Artigo em Anais de Congresso |
| Biblioteca(s): Embrapa Amazônia Oriental. |
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| 20. | | SILVA, A. R.; DIAS JUNIOR, M. de S.; LEITE, F. P.; CARVALHO, E. J. M.; VELOSO, C. A. C. Avaliação do efeito da camada de resíduos florestais na pressão de preconsolidação de um latossolo vermelho. In: CONGRESSO BRASILEIRO DE CIÊNCIA DO SOLO, 33., 2011, Uberlândia. Solos nos biomas brasileiros: sustentabilidade e mudanças climáticas: anais. [Uberlândia]: SBCS: UFU, ICIAG, 2011.| Tipo: Artigo em Anais de Congresso |
| Biblioteca(s): Embrapa Amazônia Oriental. |
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| Registros recuperados : 55 | |
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| Nenhum registro encontrado para a expressão de busca informada. |
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