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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 | |
| 7. |  | ANDREOTTI, R.; GARCIA, M. V.; CUNHA, R. C.; BARROS, J. C. Action of Tagetes minuta (Asteraceae) essential oil in the control of Rhipicephalus microplus (Acari: ixodidae). In: INTERNATIONAL SYMPOSIUM ON TICK CONTROL AND TICK-BORNE DISEASES, 2013, Campo Grande, MS. [Proceedings..]. Campo Grande, MS: Embrapa Gado de Corte, 2013. (Embrapa Gado de Corte. Documentos, 202). Comissão organizadora: Renato Andreotti, Fernando Paiva, Wilson Werner Koller, Jacqueline Cavalcante Barros, Luiz Antonio Dias Leal, Jaqueline Matias.| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Gado de Corte. |
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| 8. | | 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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| 9. | | ANDREOTTI, R.; GARCIA, M. V.; CUNHA, R. C.; BARROS, J. C. Protective action of Tagetes minuta (Asteraceae) essential oil in the control of Rhipicephalus microplus (Canestrini, 1887) (Acari: Ixodidae) in a cattle pen trial. Veterinary Parasitology, v.197, n. 1-2, out., 2013, p. 341-345| Tipo: Artigo em Periódico Indexado | Circulação/Nível: A - 1 |
| Biblioteca(s): Embrapa Gado de Corte. |
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| 10. | | 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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| 11. | | CUNHA, R. C.; ANDREOTTI, R.; SILVA, E. A. e; ELISÂNGELA PEREIRA; SATO, T.; THOMAZ-SOCCOL, V. Laboratory diagnosis and clinical signs of canine visceral leishmaniasis in dogs examined at the center for zoonosis control in Campo Grande - MS, Brazil. Archives of Veterinary Science, v. 17, n. 4, p. 17-26, 2012. Título em português: Diagnóstico laboratorial e sinais clínicos para Leishmaniose visceral canina em cães examinados no Centro de Controle de Zoonoses de Campo Grande - MS.| Tipo: Artigo em Periódico Indexado | Circulação/Nível: B - 1 |
| Biblioteca(s): Embrapa Gado de Corte. |
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| 12. | | GARCIA, M. V.; CUNHA, R. C.; ALMEIDA, R. F. C. de; MATIAS, J.; ANDREOTTI, R. Eficiência de acaricidas em larvas de Rhipicephalus sanguineus, com o uso do teste de pacote de larvas. In: CONGRESSO BRASILEIRO DE PARASITOLOGIA VETERINÁRIA, 17., 2012, São Luis. Parasitologia veterinária, bem estar e produção animal: anais. São Luis: Colégio Brasileiro de Parasitologia Veterinária, 2012. p. 157 PA 017| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Gado de Corte. |
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| 20. | | AGUIRRE, A. de A. R.; GARCIA, M. V.; CUNHA, R. C.; ANDREOTTI, R. Reverse vaccinology approach to develop an anti- tick vaccine. In: INTERNATIONAL SYMPOSIUM ON TICK CONTROL AND TICK-BORNE DISEASES, 2013, Campo Grande, MS. [Proceedings..]. Campo Grande, MS: Embrapa Gado de Corte, 2013. (Embrapa Gado de Corte. Documentos, 202). Comissão organizadora: Renato Andreotti, Fernando Paiva, Wilson Werner Koller, Jacqueline Cavalcante Barros, Luiz Antonio Dias Leal, Jaqueline Matias.| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Gado de Corte. |
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| Registros recuperados : 55 | |
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| Nenhum registro encontrado para a expressão de busca informada. |
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