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Registro Completo |
Biblioteca(s): |
Embrapa Gado de Corte. |
Data corrente: |
22/03/2011 |
Data da última atualização: |
09/09/2014 |
Tipo da produção científica: |
Artigo em Anais de Congresso |
Autoria: |
ANDREOTTI, R.; SOARES, M.; CUNHA, R. C.; LEITE, F. P. L. |
Afiliação: |
RENATO ANDREOTTI E SILVA, CNPGC; UFPEL; UFPEL; UFPEL. |
Título: |
Heterologous expression in Pichia pastoris of gene BMTI optimized. |
Ano de publicação: |
2010 |
Fonte/Imprenta: |
In: INTERNATIONAL CONGRESS OF PARASITOLOGY, 12., 2010, Melbourne. Conference abstracts. Melbourne: The Australian Society for Parasitology:World Federation of Parasitologists, 2010. |
Páginas: |
p.149-152 |
Idioma: |
Inglês Português |
Notas: |
ICOPA XII |
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 |
Thesagro: |
Carrapato; Sanidade Animal. |
Thesaurus Nal: |
Rhipicephalus microplus. |
Categoria do assunto: |
H Saúde e Patologia L Ciência Animal e Produtos de Origem Animal |
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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Embrapa Gado de Corte (CNPGC) |
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Registro Completo
Biblioteca(s): |
Embrapa Recursos Genéticos e Biotecnologia. |
Data corrente: |
08/08/2005 |
Data da última atualização: |
12/05/2022 |
Autoria: |
BRONDANI, R. P. V.; GRATTAPAGLIA, D. |
Título: |
Comparative mapping in Eucalyptus reveals significant RAPD locus linkage and order conservation between trees and greater meiotic recombination in females. |
Ano de publicação: |
1997 |
Fonte/Imprenta: |
In: IUFRO CONFERENCE ON SILVICULTURE AND IMPROVEMENT EUCALYPTS=CONFERÊNCIA IUFRO SOBRE SILVICULTURA E MELHORAMENTO DE EUCALIPTOS, 1997, Salvador. Proceedings...=Anais... Colombo: EMBRAPA-CNPF, 1997. v. 2. |
Páginas: |
p. 24-29. |
Idioma: |
Português |
Palavras-Chave: |
RAPD. |
Thesagro: |
Eucalipto. |
Categoria do assunto: |
-- |
Marc: |
LEADER 00686nam a2200145 a 4500 001 1185924 005 2022-05-12 008 1997 bl uuuu u00u1 u #d 100 1 $aBRONDANI, R. P. V. 245 $aComparative mapping in Eucalyptus reveals significant RAPD locus linkage and order conservation between trees and greater meiotic recombination in females.$h[electronic resource] 260 $aIn: IUFRO CONFERENCE ON SILVICULTURE AND IMPROVEMENT EUCALYPTS=CONFERÊNCIA IUFRO SOBRE SILVICULTURA E MELHORAMENTO DE EUCALIPTOS, 1997, Salvador. Proceedings...=Anais... Colombo: EMBRAPA-CNPF, 1997. v. 2.$c1997 300 $ap. 24-29. 650 $aEucalipto 653 $aRAPD 700 1 $aGRATTAPAGLIA, D.
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