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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: |
28/10/1992 |
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Data da última atualização: |
09/07/2013 |
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Autoria: |
SILVA, C. M. da; ZUFFO, N. L. |
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Título: |
Cultivares de soja recomendadas para o estado de Mato Grosso do Sul, safra 1985/86. |
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Ano de publicação: |
1985 |
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Fonte/Imprenta: |
Dourados: EMBRAPA-UEPAE Dourados/Campo Grande:EMPAER, 1985. |
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Páginas: |
10p. |
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Série: |
(EMBRAPA-UEPAE Dourados/EMPAER. Recomendação Técnica Conjunta, 1). |
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Idioma: |
Português |
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Conteúdo: |
Cultivares de ciclo precoce; Cultivares de ciclo médio; Cultivares de ciclo semitardio; Cultivares de ciclo tardio. |
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Palavras-Chave: |
Brasil; Cultivar; Mato Grosso do Sul; Recomendação; Soybean. |
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Thesagro: |
Cerrado; Soja; Variedade. |
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Thesaurus NAL: |
Brazil; varieties. |
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Categoria do assunto: |
--
A Sistemas de Cultivo |
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Marc: |
LEADER 00856nam a2200265 a 4500
001 1233386
005 2013-07-09
008 1985 bl uuuu u0uu1 u #d
100 1 $aSILVA, C. M. da
245 $aCultivares de soja recomendadas para o estado de Mato Grosso do Sul, safra 1985/86.
260 $aDourados: EMBRAPA-UEPAE Dourados/Campo Grande:EMPAER$c1985
300 $a10p.
490 $a(EMBRAPA-UEPAE Dourados/EMPAER. Recomendação Técnica Conjunta, 1).
520 $aCultivares de ciclo precoce; Cultivares de ciclo médio; Cultivares de ciclo semitardio; Cultivares de ciclo tardio.
650 $aBrazil
650 $avarieties
650 $aCerrado
650 $aSoja
650 $aVariedade
653 $aBrasil
653 $aCultivar
653 $aMato Grosso do Sul
653 $aRecomendação
653 $aSoybean
700 1 $aZUFFO, N. L.
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