Plasmid instability when the hsp60 gene promoter is used to express the protective non-toxic fragment B of the diphtheria toxin in recombinant BCG

Detalhes bibliográficos
Autor(a) principal: Nascimento, Dilzamar V.
Data de Publicação: 2013
Outros Autores: Dellagostin, Odir A., Hirata Jr., Raphael, Pereira, Geraldo M. B., Guaraldi, Ana Luiza Mattos, Armôa, Geraldo R. Garcia
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/12531
Resumo: Universidade do Estado do Rio de Janeiro. Faculdade de Ciências Médicas. Rio de Janeiro, RJ, Brasil.
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spelling Nascimento, Dilzamar V.Dellagostin, Odir A.Hirata Jr., RaphaelPereira, Geraldo M. B.Guaraldi, Ana Luiza MattosArmôa, Geraldo R. Garcia2016-01-14T14:19:31Z2016-01-14T14:19:31Z2013NASCIMENTO, Dilzamar V.; et al. Plasmid instability when the hsp60 gene promoter is used to express the protective non-toxic fragment B of the diphtheria toxin in recombinant BCG . American Journal of Molecular Biology, v.3, n.2, p.81-86, April 2013.2161-6620https://www.arca.fiocruz.br/handle/icict/1253110.4236/ajmb.2013.320112161-6663engScientific Research PublishingPlasmid instability when the hsp60 gene promoter is used to express the protective non-toxic fragment B of the diphtheria toxin in recombinant BCGinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleUniversidade do Estado do Rio de Janeiro. Faculdade de Ciências Médicas. Rio de Janeiro, RJ, Brasil.Universidade Federal de Pelotas. Campus Universitário. Centro de Biotecnologia. Pelotas, RS, 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.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 genetic modification of the live attenuated Mycobacterium bovis BCG to deliver a protective Corynebacterium diphtheriae antigen in vivo could be a safer and less costly alternative to the new and more expensive DTP vaccines available today, in particular to third world-countries. The stability of expression of heterologous antigens in BCG, however, is a major challenge to the use of live recombinant bacteria in vaccine development and appears to be dependent to a certain extent, on a genetic compatibility between the expression cassette within the plasmid construct and the mycobacterium host. In the quest for the best recombinant BCG transformant to express the dtb gene of C. diphtheriae we generated two new rBCG strains by transforming the Moreau substrain of BCG with the mycobacterial expression vectors pUS973 and pUS977, each one carrying a different promoter to drive the expression of the target antigen. After transformation recombinant BCG clones were selected on Middlebrook 7H10 kanamycin Agar plates, expanded in Middlebrook 7H9 kanamycin Broth and analyzed by agarose gel electrophoresis and immunoblotting. rBCGs transformed with the construct carrying the weak PAN promoter from M. paratuberculosis stably expressed the dtb gene. Conversely, rBCGs transformed with the construct carrying the strong mycobacterium hsp60 promoter were unstable and consequently unfit for the expression of the C. diphtheriae gene.BCG VaccineRecombinant BCGFragment B of Diphtheria ToxinAnti-Diphtheria VaccineVacina BCGCorynebacterium diphtheriaeDifteriaVacinas contra Difteria, Tétano e Coqueluche Acelularinfo: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/12531/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALgeraldo_armoa_etal_IOC_2013.pdfgeraldo_armoa_etal_IOC_2013.pdfapplication/pdf527419https://www.arca.fiocruz.br/bitstream/icict/12531/2/geraldo_armoa_etal_IOC_2013.pdf4ca982773f35a2eb1ee02d2129343b23MD52TEXTgeraldo_armoa_etal_IOC_2013.pdf.txtgeraldo_armoa_etal_IOC_2013.pdf.txtExtracted 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dc.title.pt_BR.fl_str_mv Plasmid instability when the hsp60 gene promoter is used to express the protective non-toxic fragment B of the diphtheria toxin in recombinant BCG
title Plasmid instability when the hsp60 gene promoter is used to express the protective non-toxic fragment B of the diphtheria toxin in recombinant BCG
spellingShingle Plasmid instability when the hsp60 gene promoter is used to express the protective non-toxic fragment B of the diphtheria toxin in recombinant BCG
Nascimento, Dilzamar V.
BCG Vaccine
Recombinant BCG
Fragment B of Diphtheria Toxin
Anti-Diphtheria Vaccine
Vacina BCG
Corynebacterium diphtheriae
Difteria
Vacinas contra Difteria, Tétano e Coqueluche Acelular
title_short Plasmid instability when the hsp60 gene promoter is used to express the protective non-toxic fragment B of the diphtheria toxin in recombinant BCG
title_full Plasmid instability when the hsp60 gene promoter is used to express the protective non-toxic fragment B of the diphtheria toxin in recombinant BCG
title_fullStr Plasmid instability when the hsp60 gene promoter is used to express the protective non-toxic fragment B of the diphtheria toxin in recombinant BCG
title_full_unstemmed Plasmid instability when the hsp60 gene promoter is used to express the protective non-toxic fragment B of the diphtheria toxin in recombinant BCG
title_sort Plasmid instability when the hsp60 gene promoter is used to express the protective non-toxic fragment B of the diphtheria toxin in recombinant BCG
author Nascimento, Dilzamar V.
author_facet Nascimento, Dilzamar V.
Dellagostin, Odir A.
Hirata Jr., Raphael
Pereira, Geraldo M. B.
Guaraldi, Ana Luiza Mattos
Armôa, Geraldo R. Garcia
author_role author
author2 Dellagostin, Odir A.
Hirata Jr., Raphael
Pereira, Geraldo M. B.
Guaraldi, Ana Luiza Mattos
Armôa, Geraldo R. Garcia
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv Nascimento, Dilzamar V.
Dellagostin, Odir A.
Hirata Jr., Raphael
Pereira, Geraldo M. B.
Guaraldi, Ana Luiza Mattos
Armôa, Geraldo R. Garcia
dc.subject.en.pt_BR.fl_str_mv BCG Vaccine
Recombinant BCG
Fragment B of Diphtheria Toxin
Anti-Diphtheria Vaccine
topic BCG Vaccine
Recombinant BCG
Fragment B of Diphtheria Toxin
Anti-Diphtheria Vaccine
Vacina BCG
Corynebacterium diphtheriae
Difteria
Vacinas contra Difteria, Tétano e Coqueluche Acelular
dc.subject.decs.pt_BR.fl_str_mv Vacina BCG
Corynebacterium diphtheriae
Difteria
Vacinas contra Difteria, Tétano e Coqueluche Acelular
description Universidade do Estado do Rio de Janeiro. Faculdade de Ciências Médicas. Rio de Janeiro, RJ, Brasil.
publishDate 2013
dc.date.issued.fl_str_mv 2013
dc.date.accessioned.fl_str_mv 2016-01-14T14:19:31Z
dc.date.available.fl_str_mv 2016-01-14T14:19:31Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
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status_str publishedVersion
dc.identifier.citation.fl_str_mv NASCIMENTO, Dilzamar V.; et al. Plasmid instability when the hsp60 gene promoter is used to express the protective non-toxic fragment B of the diphtheria toxin in recombinant BCG . American Journal of Molecular Biology, v.3, n.2, p.81-86, April 2013.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/12531
dc.identifier.issn.none.fl_str_mv 2161-6620
dc.identifier.doi.none.fl_str_mv 10.4236/ajmb.2013.32011
dc.identifier.eissn.none.fl_str_mv 2161-6663
identifier_str_mv NASCIMENTO, Dilzamar V.; et al. Plasmid instability when the hsp60 gene promoter is used to express the protective non-toxic fragment B of the diphtheria toxin in recombinant BCG . American Journal of Molecular Biology, v.3, n.2, p.81-86, April 2013.
2161-6620
10.4236/ajmb.2013.32011
2161-6663
url https://www.arca.fiocruz.br/handle/icict/12531
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language eng
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dc.publisher.none.fl_str_mv Scientific Research Publishing
publisher.none.fl_str_mv Scientific Research Publishing
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