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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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 | Acesso ao texto completo restrito à biblioteca da Embrapa Agropecuária Oeste. Para informações adicionais entre em contato com cpao.biblioteca@embrapa.br. |
Registro Completo
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Biblioteca(s): |
Embrapa Agropecuária Oeste. |
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Data corrente: |
16/07/2001 |
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Data da última atualização: |
16/07/2001 |
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Autoria: |
STAUT, L. A.; KURIHARA, C. H.; ROLIM, M. V.; ANDRADE, P. J. M. |
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Título: |
Parcelamento da cobertura nitrogenada do algodoeiro no plantio direto. |
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Ano de publicação: |
2000 |
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Fonte/Imprenta: |
In: EMBRAPA AGROPECUARIA OESTE (Dourados, MS). Resultados de pesquisa com algodao, milho e soja - safra 1999/2000: convenio Embrapa Agropecuaria Oeste e Fundacao Chapadao. Dourados, 2000. |
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Páginas: |
p.15-17 |
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Série: |
(Embrapa Agropecuaria Oeste. Documentos, 29). |
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Idioma: |
Português |
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Palavras-Chave: |
Brasil; Chapadao do Sul; Direct sowing; Fertilization; Mato Grosso do Sul. |
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Thesagro: |
Adubação; Algodão; Cerrado; Gossypium Hirsutum; Nitrogênio; Plantio Direto; Rendimento. |
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Thesaurus NAL: |
Brazil; cotton; nitrogen; yields. |
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Categoria do assunto: |
-- |
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Marc: |
LEADER 01085naa a2200361 a 4500
001 1243586
005 2001-07-16
008 2000 bl uuuu u00u1 u #d
100 1 $aSTAUT, L. A.
245 $aParcelamento da cobertura nitrogenada do algodoeiro no plantio direto.
260 $c2000
300 $ap.15-17
490 $a(Embrapa Agropecuaria Oeste. Documentos, 29).
650 $aBrazil
650 $acotton
650 $anitrogen
650 $ayields
650 $aAdubação
650 $aAlgodão
650 $aCerrado
650 $aGossypium Hirsutum
650 $aNitrogênio
650 $aPlantio Direto
650 $aRendimento
653 $aBrasil
653 $aChapadao do Sul
653 $aDirect sowing
653 $aFertilization
653 $aMato Grosso do Sul
700 1 $aKURIHARA, C. H.
700 1 $aROLIM, M. V.
700 1 $aANDRADE, P. J. M.
773 $tIn: EMBRAPA AGROPECUARIA OESTE (Dourados, MS). Resultados de pesquisa com algodao, milho e soja - safra 1999/2000: convenio Embrapa Agropecuaria Oeste e Fundacao Chapadao. Dourados, 2000.
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