Expression and purification of the immunogenically active fragment B of the Park Williams 8 Corynebacterium diphtheriae strain toxin
| Autor(a) principal: | |
|---|---|
| Data de Publicação: | 2010 |
| Outros Autores: | , , , , , , , , , , |
| Tipo de documento: | Artigo |
| Idioma: | eng |
| Título da fonte: | Repositório Institucional da FIOCRUZ (ARCA) |
| Texto Completo: | https://www.arca.fiocruz.br/handle/icict/22876 |
Resumo: | Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos. Rio de Janeiro, RJ. Brasil / Universidade do Estado do Rio de Janeiro. Faculdade de Ciências Médicas. Rio de Janeiro, RJ, Brasil. |
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Nascimento, D. V.Lemes, E. M. B.Queiroz, J. L. S.Silva Jr., J. G.Nascimento, H. J.Silva, E. D.Hirata Jr., R.Dias, A. A. S. O.,Santos, C. S.Pereira, G. M. B.Guaraldi, A. L. MattosArmoa, G. R. G.2017-10-19T10:49:28Z2017-10-19T10:49:28Z2010NASCIMENTO, D. V. et al. Expression and purification of the immunogenically active fragment B of the Park Williams 8 Corynebacterium diphtheriae strain toxin. Brazilian Journal of Medical and Biological Research, v.43, p.460-466, 2010.0100-879Xhttps://www.arca.fiocruz.br/handle/icict/228761414-431X)engAssociação Brasileira de Divulgação CientíficaDifteriaEscherichia coliExpressão GênicaAfinidade do metal imobilizadoFragment BDiphtheriadtb geneE. coli gene expressionImmobilized metal affinityExpression and purification of the immunogenically active fragment B of the Park Williams 8 Corynebacterium diphtheriae strain toxininfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos. Rio de Janeiro, RJ. Brasil / Universidade do Estado do Rio de Janeiro. Faculdade de Ciências Médicas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos. Rio de Janeiro, RJ. Brasil.Universidade do Estado do Rio de Janeiro. Faculdade de Ciências Médicas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Controle de Qualidade em Saúde. Rio de Janeiro, RJ, Brasil.Universidade do Estado do Rio de Janeiro. Faculdade de Ciências Médicas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ. Brasil / Universidade do Estado do Rio de Janeiro. Faculdade de Ciências Médicas. Rio de Janeiro, RJ, Brasil.Universidade do Estado do Rio de Janeiro. Faculdade de Ciências Médicas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ. Brasil.The construction of a hexahistidine-tagged version of the B fragment of diphtheria toxin (DTB) represents an important step in the study of the biological properties of DTB because it will permit the production of pure recombinant DTB (rDTB) in less time and with higher yields than currently available. In the present study, the genomic DNA of the Corynebacterium diphtheriae Park Williams 8 (PW8) vaccine strain was used as a template for PCR amplification of the dtb gene. After amplification, the dtb gene was cloned and expressed in competent Escherichia coli M15™ cells using the expression vector pQE-30™. The lysate obtained from transformed E. coli cells containing the rDTBPW8 was clarified by centrifugation and purified by affinity chromatography. The homogeneity of the purified rDTBPW8 was confirmed by immunoblotting using mouse polyclonal anti-diphtheria toxoid antibodies and the immune response induced in animals with rDTBPW8 was evaluated by ELISA and dermonecrotic neutralization assays. The main result of the present study was an alternative and accessible method for the expression and purification of immunogenically reactive rDTBPW8 using commercially available systems. Data also provided preliminary evidence that rabbits immunized with rDTBPW8 are able to mount a neutralizing response against the challenge with toxigenic C. diphtheriaeinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-82991https://www.arca.fiocruz.br/bitstream/icict/22876/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALGRG_armoa_etal_IOC_2010.pdfGRG_armoa_etal_IOC_2010.pdfapplication/pdf910390https://www.arca.fiocruz.br/bitstream/icict/22876/2/GRG_armoa_etal_IOC_2010.pdf94fa256eabde895e2f0f6342a63edc54MD52TEXTGRG_armoa_etal_IOC_2010.pdf.txtGRG_armoa_etal_IOC_2010.pdf.txtExtracted texttext/plain26188https://www.arca.fiocruz.br/bitstream/icict/22876/3/GRG_armoa_etal_IOC_2010.pdf.txtf5491879eef0be3e06b48d356f06d025MD53icict/228762018-08-15 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| dc.title.pt_BR.fl_str_mv |
Expression and purification of the immunogenically active fragment B of the Park Williams 8 Corynebacterium diphtheriae strain toxin |
| title |
Expression and purification of the immunogenically active fragment B of the Park Williams 8 Corynebacterium diphtheriae strain toxin |
| spellingShingle |
Expression and purification of the immunogenically active fragment B of the Park Williams 8 Corynebacterium diphtheriae strain toxin Nascimento, D. V. Difteria Escherichia coli Expressão Gênica Afinidade do metal imobilizado Fragment B Diphtheria dtb gene E. coli gene expression Immobilized metal affinity |
| title_short |
Expression and purification of the immunogenically active fragment B of the Park Williams 8 Corynebacterium diphtheriae strain toxin |
| title_full |
Expression and purification of the immunogenically active fragment B of the Park Williams 8 Corynebacterium diphtheriae strain toxin |
| title_fullStr |
Expression and purification of the immunogenically active fragment B of the Park Williams 8 Corynebacterium diphtheriae strain toxin |
| title_full_unstemmed |
Expression and purification of the immunogenically active fragment B of the Park Williams 8 Corynebacterium diphtheriae strain toxin |
| title_sort |
Expression and purification of the immunogenically active fragment B of the Park Williams 8 Corynebacterium diphtheriae strain toxin |
| author |
Nascimento, D. V. |
| author_facet |
Nascimento, D. V. Lemes, E. M. B. Queiroz, J. L. S. Silva Jr., J. G. Nascimento, H. J. Silva, E. D. Hirata Jr., R. Dias, A. A. S. O., Santos, C. S. Pereira, G. M. B. Guaraldi, A. L. Mattos Armoa, G. R. G. |
| author_role |
author |
| author2 |
Lemes, E. M. B. Queiroz, J. L. S. Silva Jr., J. G. Nascimento, H. J. Silva, E. D. Hirata Jr., R. Dias, A. A. S. O., Santos, C. S. Pereira, G. M. B. Guaraldi, A. L. Mattos Armoa, G. R. G. |
| author2_role |
author author author author author author author author author author author |
| dc.contributor.author.fl_str_mv |
Nascimento, D. V. Lemes, E. M. B. Queiroz, J. L. S. Silva Jr., J. G. Nascimento, H. J. Silva, E. D. Hirata Jr., R. Dias, A. A. S. O., Santos, C. S. Pereira, G. M. B. Guaraldi, A. L. Mattos Armoa, G. R. G. |
| dc.subject.other.pt_BR.fl_str_mv |
Difteria Escherichia coli Expressão Gênica Afinidade do metal imobilizado |
| topic |
Difteria Escherichia coli Expressão Gênica Afinidade do metal imobilizado Fragment B Diphtheria dtb gene E. coli gene expression Immobilized metal affinity |
| dc.subject.en.pt_BR.fl_str_mv |
Fragment B Diphtheria dtb gene E. coli gene expression Immobilized metal affinity |
| description |
Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos. Rio de Janeiro, RJ. Brasil / Universidade do Estado do Rio de Janeiro. Faculdade de Ciências Médicas. Rio de Janeiro, RJ, Brasil. |
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2010 |
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2010 |
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2017-10-19T10:49:28Z |
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2017-10-19T10:49:28Z |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/article |
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article |
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NASCIMENTO, D. V. et al. Expression and purification of the immunogenically active fragment B of the Park Williams 8 Corynebacterium diphtheriae strain toxin. Brazilian Journal of Medical and Biological Research, v.43, p.460-466, 2010. |
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https://www.arca.fiocruz.br/handle/icict/22876 |
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0100-879X |
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1414-431X) |
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NASCIMENTO, D. V. et al. Expression and purification of the immunogenically active fragment B of the Park Williams 8 Corynebacterium diphtheriae strain toxin. Brazilian Journal of Medical and Biological Research, v.43, p.460-466, 2010. 0100-879X 1414-431X) |
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Associação Brasileira de Divulgação Científica |
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